KR20190142456A - 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 - Google Patents
인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 Download PDFInfo
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Abstract
인시류 저해 활성을 나타내는 신규한 키메릭 살곤충 단백질(chimeric insecticidal protein)을 암호화하는 뉴클레오타이드 서열이 개시된다. 특정 실시형태는 키메릭 살곤충 단백질 중 1종 이상을 암호화하는 재조합 핵산 분자를 함유하는 조성물 및 형질전환된 식물, 식물 부분 및 종자를 제공한다.
Description
관련 출원에 대한 참고
본 출원은 2014년 10월 16일자로 출원된 미국 가출원 제62/064,989호의 이익을 청구하며, 그것은 전문이 참고로 본원에 포함된다.
서열 목록의 포함
컴퓨터 판독가능한 형태의 서열 목록이 전자 제출에 의해서 본 출원과 함께 제출되고, 그의 전체가 참고로 본 출원에 포함된다. 서열 목록은 371,735 킬로바이트 크기(MS-윈도우즈(MS-Windows)(등록상표) 운용 시스템에서 측정)이고, 파일명이 P34230WO00_SEQ_PCT.txt인 2015년 10월 13일자로 생성된 파일에 포함되어 있다.
기술 분야
본 발명은 일반적으로 곤충 저해 단백질의 분야에 관한 것이다. 작물 및 종자의 농업-관련 해충에 대한 곤충 저해 활성을 나타내는 신규한 부류의 키메릭 살곤충 단백질(chimeric insecticidal protein)이 본 출원에서 개시된다. 특히, 개시된 부류의 단백질은 인시목(Lepidopteran order)의 곤충 해충에 대해서 살곤충 활성을 나타낸다. 개시된 독소(toxin) 단백질 중 하나 이상을 암호화하는 재조합 핵산 분자를 함유하는 식물, 식물 부분, 및 종자가 제공된다.
특히, 옥수수, 대두, 사탕수수, 벼, 밀, 채소류 및 목화를 비롯한 농업에서 중요한 식물로부터의 작물 수확량의 개선이 상당히 중요해지고 있다. 식량에 대한 농업 생산물에 대한 증가하고 있는 요구에 더하여, 의류 및 인구 증가에 따른 에너지 제공, 기후 관련 효과 및 농사 이외의 용도로 토지를 사용하려는 인구 증가로부터의 압력은 농사를 위해 사용 가능한 경작지의 양을 감소시킬 것으로 예상된다. 이러한 인자는 특히 식물 생명공학 및 농경법 실시에서의 상당한 개선이 없이는, 식량 안보와 관련하여 암울한 예측으로 이어진다. 이러한 압력에 비추어, 기술, 농업 기술 및 해충 제어에서의 환경적으로 지속가능한 개선이 농사에 사용 가능한 제한된 경작지 면적에 대한 작물 생산성 확대를 위한 필수적인 툴이다.
곤충, 특히 인시목의 곤충은 농작물 손실의 주요 원인인 것으로 간주되어, 침입된 지역에서 작물 생산량을 감소시킨다. 농업에 부정적으로 영향을 미치는 인시류 해충 종은 가을멸강충(fall armyworm)(스포돕테라 프루기페르다(Spodoptera frugiperda)), 파밤나방(beet armyworm)(스포돕테라 엑시구아(Spodoptera exigua)), 베르타 밤나방(bertha armyworm)(마메스트라 콘피구라타(Mamestra configurata)), 검거세미 나방(black cutworm)(아그로티스 입실론(Agrotis ipsilon)), 남방은무늬밤나방 애벌레(cabbage looper)(트리코플루시아 니(Trichoplusia ni)), 대두 애벌레(soybean looper)(크리소데익시스 인클루덴스(Chrysodeixis includens)), 벨벳콩 자나방(velvetbean caterpillar)(안티카르시아 겜마탈리스(Anticarsia gemmatalis)), 그린 클로버웜(green cloverworm)(히페나 스카브라(Hypena scabra)), 회색 담배 나방(tobacco budworm)(헬리오티스 비레센스(Heliothis virescens)), 낟알 거세미 나방(granulate cutworm)(아그로티스 수브테라네아(Agrotis subterranea)), 멸강나방(armyworm)(수달레티아 유니펀크타(Pseudaletia unipuncta)), 서양 거세미 나방(western cutworm)(아그로티스 오르토고니아(Agrotis orthogonia)), 유럽 조명충 나방(European corn borer)(오스트리니아 누빌랄리스(Ostrinia nubilalis)), 네이블 오렌지 나방(navel orangeworm)(아미엘로이스 트란시텔라(Amyelois transitella)), 옥수수 뿌리 벌집 나방(corn root webworm)(크람부스 칼리기노셀루스(Crambus caliginosellus)), 소드 벌집 나방(sod webworm)(헤르페토그라마 리카르시살리스(Herpetogramma licarsisalis)), 해바라기 나방(sunflower moth)(호모에오소마 엘렉텔룸(Homoeosoma electellum)), 명충 나방 애벌레(lesser cornstalk borer)(엘라스모팔푸스 리그노셀루스(Elasmopalpus lignosellus)), 코들링 나방(codling moth)(시디아 포모넬라(Cydia pomonella)), 그레이프 베리 나방(grape berry moth)(엔도피자 비테아나(Endopiza viteana)), 복숭아순 나방(oriental fruit moth)(그라폴리타 몰레스타(Grapholita molesta)), 해바라기순 나방(sunflower bud moth)(술레이마 헬리안타나(Suleima helianthana)), 배추좀 나방(diamondback moth)(플루텔라 크실로스텔라(Plutella xylostella)), 분홍 솜벌레(pink bollworm)(펙티노포라 고시피엘라(Pectinophora gossypiella)), 분홍 명밤 나비(pink stem borer)(세사미아 인페렌스(Sesamia inferens)), 매미 나방(gypsy moth)(리만트리아 디스파르(Lymantria dispar)), 목화 잎 벌레(cotton leaf worm)(알라바마 아르길라세아(Alabama argillacea)), 과일 나무 잎말이 나방(fruit tree leaf roller)(아르킵스 아르기로스필라(Archips argyrospila)), 유럽 잎말이 나방(European leafroller)(아르킵스 로사나(Archips rosana)), 이화명 나방(Asiatic rice borer), 또는 쌀 명밤 나방(rice stem borer)(킬로 서프레살리스(Chilo suppressalis)), 혹명 나방(rice leaf roller)(크나팔로크로시스 메디날리스(Cnaphalocrocis medinalis)), 옥수수 뿌리 벌집 나방(크람부스 칼리기노셀루스(Crambus caliginosellus)), 잔디 포충 나방(bluegrass webworm)(크람부스 테테렐루스(Crambus teterrellus)), 남서부 조명충 나방(southwestern corn borer)(디아트라에아 그란디오셀라(Diatraea grandiosella)), 사탕수수 명나방(surgarcane borer)(디아트라에아 사카랄리스(Diatraea saccharalis)), 스피니 볼웜(spiny bollworm)(에아리아스 인술라나(Earias insulana)), 스팟티드 볼웜(spotted bollworm)(이아리아스 비텔라(Earias vittella)), 구세계 목화씨 벌레(Old World cotton bollworm)(헬리코베르파 아르미게라(Helicoverpa armigera)), 왕담배 밤나방(corn earworm), 콩 팟웜(soy podworm) 또는 목화씨 벌레(헬리코베르파 제아(Helicoverpa zea)), 소드 벌집 나방(헤르페토그라마 리카르시살리스(Herpetogramma licarsisalis)), 유럽 포도나무 나방(European grape vine moth)(로베시아 보트라나(Lobesia botrana)), 귤굴 나방(citrus leafminer)(필록니스티스 시트렐라(Phyllocnistis citrella)), 큰 흰나비(large white butterfly)(피에리스 브라시카에(Pieris brassicae)), 배추 흰나비(imported cabbageworm), 또는 작은 흰나비(피에리스 라파에(Pieris rapae)), 담배 거세미 나방(tobacco cutworm), 또는 클러스터 캐터필라(cluster caterpillar)(스포돕테라 리투라(Spodoptera litura)), 및 토마토 잎나방(tomato leafminer)(투타 앱솔루타(Tuta absoluta))을 포함하지만, 이들로 제한되는 것은 아니다.
역사적으로, 합성 화학 살곤충제의 집중적인 살포가 농업에서 해충 방제제로서 신뢰되었다. 저항성 문제의 출현에 더하여, 환경 및 인간 건강에 대한 우려가 생물학적 살충제의 연구 및 개발을 자극하였다. 이러한 연구 노력은 박테리아를 비롯한, 다양한 곤충병원성 미생물 종의 점진적인 발견 및 사용으로 이어졌다.
곤충병원성 박테리아, 특히 바실러스속에 속하는 박테리아의 생물학적 해충 방제제로서의 가능성이 발견되고, 발전되었을 때 생물학적 방제 패러다임이 이동하였다. 박테리아 바실러스 투린기엔시스(Bt)의 균주를 살곤충 단백질을 위한 공급원으로서 사용하여 왔는데, 그 이유는 Bt 균주가 특정 곤충에 대해서 높은 독성을 나타내는 것을 발견하였기 때문이다. Bt 균주는 포자 형성의 시작 시에 그리고 성장 정체기(stationary growth phase) 동안 부아포 결정 봉입체(parasporal crystalline inclusion body) 내에 국지화된 델타-엔도톡신(예를 들어, Cry 단백질)을 생성한다고 공지되어 있으며, 분비된 살곤충 단백질을 생성한다고 또한 공지되어 있다. 민감한 곤충이 섭취한 후, 델타-엔도톡신뿐만 아니라 분비된 독소는 중장 상피(midgut epithelium)의 표면에서 그의 효과를 발휘하여, 세포막을 파괴하여, 세포를 파괴하고, 사멸시킨다. 살곤충 단백질을 암호화하는 유전자는 또한 다른 바실러스 및 다양한 다른 박테리아 종, 예컨대 브레비바실러스 래터로스포러스(Brevibacillus laterosporus), 리시니바실러스 스파에리커스(Lysinibacillus sphaericus)("Ls" 이전에 바실러스 스파에리커스라 알려짐) 및 파에니바실러스 포필리아에(Paenibacillus popilliae)에서 확인되었다.
결정성인 분비된 가용성 살곤충 단백질 독소는 그의 숙주에 대해서 고도로 특이적이어서, 화학적 살곤충제에 대한 대체품으로서 전세계적으로 허용되어 왔다. 예를 들어, 살곤충 독소 단백질은 다양한 농업 응용에서 사용되어 농업적으로 중요한 식물을 곤충 침입으로부터 보호하고, 화학적 살충제 적용에 대한 요구를 감소시키고, 수확량을 증가시켰다. 살곤충 독소 단백질은 기계적 방법, 예컨대 분무하여 다양한 박테리아 균주를 함유하는 미생물 제제를 식물 표면 상에 확산시킴으로써, 그리고 유전자 형질전환 기술(genetic transformation technique)을 사용하여 살곤충 독소 단백질을 발현하는 트랜스제닉 식물(transgenic plant) 및 종자를 생산함으로써 작물의 농업 관련 해충을 방제하는 데 사용된다.
살곤충 단백질을 발현하는 트랜스제닉 식물의 사용은 전세계적으로 채택되어 왔다. 예를 들어, 2012년에, 2천6백십만 헥타아르에 Bt 독소를 발현하는 트랜스제닉 작물을 심었다(문헌 [James, C., Global Status of Commercialized Biotech/GM Crops: 2012. ISAAA Brief No. 44]). 트랜스제닉 곤충-보호된 작물의 전세계적 사용 및 이러한 작물에서 사용된 살곤충 단백질의 제한된 수가 현재 사용되는 살곤충 단백질에 저항성을 부여하는 기존의 곤충 대립유전자에 대한 선별 압력을 생성하였다.
살곤충 단백질에 대한 표적 해충의 저항성의 발전이, 살곤충 단백질을 발현하는 트랜스제닉 작물에 대한 곤충 저항성의 증가를 조절하기에 유용한 새로운 형태의 살곤충 단백질의 발견 및 발전에 대한 계속적인 필요성을 생성한다. 효능이 개선되고, 더 넓은 스펙트럼의 허용되는 곤충 종에 대해서 방제를 나타내는 새로운 살곤충 단백질이 저항성 대립유전자를 발전시킬 수 있는 살아남은 곤충의 수를 감소시킬 것이다. 또한, 동일한 곤충 해충에 독성이 있고, 상이한 작용 기전을 나타내는 2종 이상의 트랜스제닉 살곤충 단백질을 한 식물에서 사용하는 것이 임의의 단일 표적 곤충 종에서 저항성 확률을 감소시킨다.
결론적으로, 농업 분야에서 현재 사용되는 독소에 비해서 개선된 살곤충 특성, 예컨대 더 넓은 스펙트럼의 표적 곤충 해충 종에 대한 증가된 효능 및 상이한 작용 기전을 갖는 추가적인 살곤충 단백질을 연구하는 것이 상당히 필요하다. 이러한 필요성을 충족시키기 위해서, 본 발명은 중요한 표적 인시류 해충 종에 대해서 활성을 나타내는 신규한 Cry1 키메릭 살곤충 단백질을 개시한다.
Cry1 결정 단백질 군의 구성원은 인시류 해충에 대해서 생물활성을 나타낸다고 당업계에 공지되어 있다. Cry1 결정 단백질의 전구체 형태는 2개의 대략 동일한 크기의 분절로 이루어진다. 프로톡신(protoxin) 분절로서 공지된, 전구체 단백질의 카르복시-말단 부분은 결정 형성을 안정화시키고, 살곤충 활성을 나타내지 않는다. 전구체 단백질의 아미노 말단 절반부는 Cry1 단백질의 독소 분절을 포함하고, Cry1 군 구성원 내에 보존되거나 실질적으로 보존된 서열의 정렬에 기초하여, 3개의 구조 도메인, 즉 도메인 I, 도메인 II, 및 도메인 III로 추가로 나뉠 수 있다. 도메인 I은 활성 독소 분절의 대략 1/3을 포함하고, 채널 형성에 필수적인 것으로 밝혀져 있다. 도메인 II 및 III 모두는 연구될 곤충 및 살곤충 단백질에 따라서, 수용체 결합 및 곤충 종 특이성에 관련된다.
당업계에서 공지된 많은 자연 존재(native) 살곤충 단백질의 도메인 구조의 조합으로부터 향상된 특성을 갖는 키메릭 단백질을 임의로 생성할 가능성은 희박하다. 이것은 단백질 구조, 올리고머화, 및 살곤충 단백질 분절을 방출시키는 데 요구되는 활성화(그러한 형태로 발현된다면, 키메릭 전구체의 올바른 단백질 가수분해 가공 포함)의 복잡한 성질의 결과이다. 키메릭 구조의 생성을 위해서 각각의 양친(parental) 단백질 내에서 프로톡신 및 특이적인 타겟을 주의깊게 선택하는 것 만이 기능성 키메릭 살곤충 독소가 키메라가 유래된 양친 단백질에 비해서 개선된 살곤충 활성을 나타내도록 구성될 수 있게 한다. 서로 상이한 임의의 둘 이상의 독소의 프로톡신 및 독소 도메인 I, II 및 III의 재조립은 보통 잘못된 결정 형성을 나타내거나, 바람직한 표적 곤충 해충 종에 대해서 유도되는 임의의 검출가능한 살곤충 활성이 완전히 결핍된 단백질 구성을 유발한다고 당업계에 공지되어 있다. 시행 착오를 거쳐서 만이 효과적인 살곤충 키메라를 설계하였지만, 그 이후에도, 통상의 기술자는 키메라의 구성 프로톡신 또는 독소 도메인이 유래될 수 있는 임의의 단일 양친 독소 단백질과 동등하거나 또는 그것에 비해서 개선된 살곤충 활성을 나타내는 키메라를 설계했다고 확신하지 못한다. 예를 들어, 문헌은 2종 이상의 결정 단백질 전구체로부터의 키메릭 단백질의 구성 또는 조립의 다양한 예를 보고한다. 예를 들어, 문헌[Jacqueline S. Knight, et al. "A Strategy for Shuffling Numerous Bacillus thuringiensis Crystal Protein Domains." J. Economic Entomology, 97 (6) (2004): 1805-1813]; 보쉬(Bosch) 등의 특허(미국 특허 제6,204,246호); 말바(Malvar) 및 길머(Gilmer)의 특허(미국 특허 제6,017,534호)를 참고하기 바란다. 이들 예 각각에서, 생성된 키메라 중 다수는 키메라의 성분이 유래된 전구체 단백질과 동등하거나 또는 그에 비해서 개선된 살곤충 또는 결정 형성 특성을 나타내지 않았다.
인시류 종의 식물 해충에 유독성인 키메릭 살곤충 단백질을 암호화하는 재조합 핵산 분자가 제공된다. 키메릭 살곤충 단백질 각각은 제제 중에서 그리고 식물체 내에서 단독으로 또는 서로와 그리고 다른 살곤충 단백질 및 곤충 저해제와 조합되어 사용될 수 있기 때문에; 농업계에서 현재 사용되는 살곤충 단백질 및 살곤충 화학물질에 대한 대안을 제공한다.
특정 실시형태에서, 본원에는 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 바와 같은 아미노산 서열을 포함하는 키메릭 살곤충 단백질이 개시되어 있다. 이러한 키메릭 살곤충 단백질은 인시목 곤충 종, 예컨대 안티카르시아 겜마탈리스, 디아트라에아 사카랄리스, 엘라스모팔푸스 리그노셀루스, 헬리코베르파 제아, 헬리오티스 비레센스, 크리소데익시스 인클루덴스, 스포돕테라 코스미오이데스(Spodoptera cosmioides), 스포돕테라 에리다니아(Spodoptera eridania), 스포돕테라 프루기페르다, 스포돕테라 엑시구아, 헬리코베르파 아르미게라, 스포돕테라 리투라, 펙티노포라 고시피엘라, 디아트라에아 그란디오셀라, 에아리아스 비텔라, 헬리코베르파 겔로토페온, 및 라치플루시아 누(Rachiplusia nu) (이에 제한되지 않음)에 대해서 저해 활성을 나타낸다.
또 다른 실시형태에서, 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드가 개시되며, 여기서 폴리뉴클레오타이드는 이종 프로모터에 작동 가능하게 연결되어 있고, 키메릭 살곤충 단백질은 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 바와 같은 아미노산 서열을 포함한다. 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드가 또한 고려되며, 여기서 폴리뉴클레오타이드는 임의로 엄격한 조건(stringent condition) 하에서 서열번호 1, 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 22, 24, 26, 27, 29, 31, 32, 34, 35, 37, 38, 40, 42, 44, 46, 48, 49, 51 또는 52 중 어느 하나에 명시된 바와 같은 폴리뉴클레오타이드 서열의 역상보체에 혼성화(hybridizing)되거나; 또는 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 바와 같은 아미노산 서열을 포함하는 키메릭 살곤충 단백질을 암호화하는 뉴클레오타이드 서열을 포함한다.
다른 실시형태에서, 본원에는 서열번호 1, 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 22, 24, 26, 27, 29, 31, 32, 34, 35, 37, 38, 40, 42, 44, 46, 48, 49, 51 또는 52 중 어느 하나에 명시된 폴리뉴클레오타이드를 포함하는 숙주 세포가 개시되어 있고, 여기서 숙주 세포는 박테리아 숙주 세포 또는 식물 숙주 세포로 이루어진 군으로부터 선택된다. 고려되는 박테리아 숙주에는 아그로박테리움(Agrobacterium), 리조븀(Rhizobium), 바실러스(Bacillus), 브레비바실러스(Brevibacillus), 에쉐리키아(Escherichia), 슈도모나스(Pseudomonas), 클렙시엘라(Klebsiella), 및 에르위니아(Erwinia)가 포함되며; 여기서 바실러스 종은 바실러스 세레우스(Bacillus cereus) 또는 바실러스 투린기엔시스(Bacillus thuringiensis)이고, 상기 브레비바실러스가 브레비바실러스 라테로스페러스(Brevibacillus laterosperous)이고, 상기 에쉐리키아가 에쉐리키아 콜라이(Escherichia coli)이다. 고려되는 식물 세포에는 외떡잎 식물 및 쌍떡잎 식물이 포함된다.
본 명세서에 개시된 다른 실시형태는 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 바와 같은 아미노산 서열을 포함하는 키메릭 살곤충 단백질을 포함하는 곤충 저해 조성물을 포함한다. 특정 실시형태에서, 곤충 저해 조성물은 키메릭 살곤충 단백질과 상이한 적어도 1종의 곤충 저해제를 추가로 포함한다. 키메릭 살곤충 단백질과 상이한 고려되는 곤충 저해제에는 곤충 저해 단백질, 곤충 저해 dsRNA 분자, 및 곤충 저해 화학물질이 포함된다. 키메릭 살곤충 단백질과 상이한 이러한 곤충 저해제는 인시목, 딱정벌레목, 노린재목, 동시아목, 또는 총채벌레목 중 1종 이상의 해충 종에 대해서 활성을 나타낸다.
또 다른 실시형태에서, 본원에는 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 바와 같은 아미노산 서열을 포함하는 키메릭 살곤충 단백질; 또는 서열번호 1, 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 22, 24, 26, 27, 29, 31, 32, 34, 35, 37, 38, 40, 42, 44, 46, 48, 49, 51 또는 52 중 어느 하나에 명시된 폴리뉴클레오타이드를 곤충 저해 유효량으로 포함하는 종자가 개시되어 있다.
인시류 해충을 억제량의 본 발명의 키메릭 살곤충 단백질과 접촉시키는 것을 포함하는 인시류 해충의 방제 방법이 또한 고려된다.
또 다른 실시형태에서, 본원에는 키메릭 살곤충 단백질을 포함하는 트랜스제닉 식물 세포, 식물 또는 식물 부분이 개시되어 있으며, 여기서 키메릭 살곤충 단백질은 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 임의의 아미노산 서열을 포함하거나; 또는 키메릭 살곤충 단백질은 서열번호 21, 10에 대해서 적어도 94% 동일성; 서열번호 28에 대해서 적어도 93% 동일성; 서열번호 7에 대해서 적어도 87% 동일성; 서열번호 4에 대해서 적어도 90% 동일성; 서열번호 13에 대해서 적어도 91% 동일성; 서열번호 16에 대해서 적어도 64% 동일성; 서열번호 19에 대해서 적어도 66% 동일성; 서열번호 23에 대해서 적어도 86% 동일성; 서열번호 25에 대해서 적어도 91% 동일성; 서열번호 30에 대해서 적어도 94% 동일성; 서열번호 33에 대해서 적어도 91% 동일성; 서열번호 36에 대해서 적어도 64% 동일성; 서열번호 39에 대해서 적어도 66% 동일성; 서열번호 41에 대해서 적어도 94% 동일성; 서열번호 43에 대해서 적어도 84% 동일성; 서열번호 45에 대해서 적어도 93% 동일성; 서열번호 47에 대해서 적어도 94% 동일성; 서열번호 50에 대해서 적어도 91% 동일성; 또는 서열번호 53에 대해서 적어도 93% 동일성을 갖는 단백질을 포함한다. 인시류 해충을 이러한 트랜스제닉 식물 세포, 식물 또는 식물 부분에 노출시키는 것을 포함하는 인시류 해충의 방제 방법이 또한 고려되며, 여기서 상기 식물 세포, 식물 또는 식물 부분은 인시류 억제량의 키메릭 살곤충 단백질을 발현한다.
본원의 다른 실시형태에서, 식물 세포, 식물, 또는 식물 부분으로부터 유래된 상품(commodity product)이 제공되고, 여기서 상품은 검출가능한 양의 키메릭 살곤충 단백질을 포함한다. 고려되는 상품에는 식물 바이오매스(biomass), 오일, 곡물(meal), 동물 사료, 곡물 가루, 플레이크, 겨(bran), 린트, 외피, 및 가공된 종자가 포함된다.
본 명세서에 개시된 또 다른 방법은 키메릭 살곤충 단백질을 포함하는 종자의 생산 방법이며, 그 방법은 키메릭 살곤충 단백질을 포함하는 적어도 하나의 종자를 심는 단계; 상기 종자로부터 식물을 성장시키는 단계; 및 상기 식물로부터 종자를 수확하는 단계를 포함하고, 여기서 상기 수확된 종자는 키메릭 살곤충 단백질을 포함한다.
*1, 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 22, 24, 26, 27, 29, 31, 32, 34, 35, 37, 38, 40, 42, 44, 46, 48, 49, 51 또는 52로 이루어진 군으로부터 선택된 뉴클레오타이드 서열을 포함하는, 키메릭 살곤충 단백질을 암호화하는 재조합 폴리뉴클레오타이드 분자; 및 임의로 키메릭 살곤충 단백질과 상이한 곤충 저해제를 암호화하는 폴리뉴클레오타이드 서열이 또한 본 명세서에서 고려된다.
본 명세서에서 고려되는 또 다른 재조합 핵산 분자는 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드 분절에 작동 가능하게 연결된 이종 프로모터를 포함하고, 여기서 키메릭 살곤충 단백질은 서열번호 21, 10, 28, 7, 4, 13, 16, 19, 23, 25, 30, 33, 36, 39, 41, 43, 45, 47, 50 또는 53 중 어느 하나에 명시된 임의의 아미노산 서열을 포함하거나; 또는 키메릭 살곤충 단백질은 서열번호 21, 10에 대해서 적어도 94% 동일성; 서열번호 28에 대해서 적어도 93% 동일성; 서열번호 7에 대해서 적어도 87% 동일성; 서열번호 4에 대해서 적어도 90% 동일성; 서열번호 13에 대해서 적어도 91% 동일성; 서열번호 16에 대해서 적어도 64% 동일성; 서열번호 19에 대해서 적어도 66% 동일성; 서열번호 23에 대해서 적어도 86% 동일성; 서열번호 25에 대해서 적어도 91% 동일성; 서열번호 30에 대해서 적어도 94% 동일성; 서열번호 33에 대해서 적어도 91% 동일성; 서열번호 36에 대해서 적어도 64% 동일성; 서열번호 39에 대해서 적어도 66% 동일성; 서열번호 41에 대해서 적어도 94% 동일성; 서열번호 43에 대해서 적어도 84% 동일성; 서열번호 45에 대해서 적어도 93% 동일성; 서열번호 47에 대해서 적어도 94% 동일성; 서열번호 50에 대해서 적어도 91% 동일성; 또는 서열번호 53에 대해서 적어도 93% 동일성을 갖는 단백질을 포함하거나; 또는 폴리뉴클레오타이드 분절은 서열번호 1, 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 22, 24, 26, 27, 29, 31, 32, 34, 35, 37, 38, 40, 42, 44, 46, 48, 49, 51 또는 52 중 어느 하나에 명시된 바와 같은 뉴클레오타이드 서열을 갖는 폴리뉴클레오타이드에 혼성화된다.
본 발명의 다른 실시형태, 특징 및 이점은 하기 발명을 실시하기 위한 구체적인 내용, 실시예 및 청구범위로부터 명백할 것이다.
서열의 간단한 설명
서열번호 서열번호 1은 박테리아 세포에서의 발현을 위해서 사용되는 TIC1100을 암호화하는 재조합 DNA 서열이다.
서열번호 2는 식물 세포에서의 발현을 위한 TIC1100을 암호화하는 합성 DNA 서열이다.
서열번호 3은 식물 세포에서의 발현을 위한 TIC1100을 암호화하는 합성 DNA 서열이다.
서열번호 4는 TIC1100의 아미노산 서열이다.
서열번호 5는 박테리아 세포에서의 발현을 위해서 사용되는 TIC860을 암호화하는 재조합 DNA 서열이다.
서열번호 6은 식물 세포에서의 발현을 위한 TIC860을 암호화하는 합성 DNA 서열이다.
서열번호 7은 TIC860의 아미노산 서열이다.
서열번호 8은 박테리아 세포에서의 발현을 위해서 사용되는 TIC867을 암호화하는 재조합 DNA 서열이다.
서열번호 9는 식물 세포에서의 발현을 위한 TIC867을 암호화하는 합성 DNA 서열이다.
서열번호 10은 TIC867의 아미노산 서열이다.
서열번호 11은 박테리아 세포에서의 발현을 위해서 사용되는 TIC867_20을 암호화하는 재조합 DNA 서열이다.
서열번호 12는 식물 세포에서의 발현을 위한 TIC867_20을 암호화하는 합성 DNA 서열이다.
서열번호 13은 TIC867_20의 아미노산 서열이다.
서열번호 14는 박테리아 세포에서의 발현을 위해서 사용되는 TIC867_21을 암호화하는 재조합 DNA 서열이다.
서열번호 15는 식물 세포에서의 발현을 위한 TIC867_21을 암호화하는 합성 DNA 서열이다.
서열번호 16은 TIC867_21의 아미노산 서열이다.
서열번호 17은 박테리아 세포에서의 발현을 위해서 사용되는 TIC867_22를 암호화하는 재조합 DNA 서열이다.
서열번호 18은 식물 세포에서의 발현을 위한 TIC867_22를 암호화하는 합성 DNA 서열이다.
서열번호 19는 TIC867_22의 아미노산 서열이다.
서열번호 20은 식물 세포에서의 발현을 위한 TIC867_23을 암호화하는 합성 DNA 서열이다.
서열번호 21은 TIC867_23의 아미노산 서열이다.
서열번호 22는 식물 세포에서의 발현을 위한 TIC867_24를 암호화하는 합성 DNA 서열이다.
서열번호 23은 TIC867_24의 아미노산 서열이다.
서열번호 24는 식물 세포에서의 발현을 위한 TIC867_24를 암호화하는 합성 DNA 서열이다.
서열번호 25는 TIC867_25의 아미노산 서열이다.
서열번호 26은 박테리아 세포에서의 발현을 위해서 사용되는 TIC868을 암호화하는 재조합 DNA 서열이다.
서열번호 27은 식물 세포에서의 발현을 위한 TIC868을 암호화하는 합성 DNA 서열이다.
서열번호 28은 TIC868의 아미노산 서열이다.
서열번호 29는 식물 세포에서의 발현을 위한 TIC868_9를 암호화하는 합성 DNA 서열이다.
서열번호 30은 TIC868_9의 아미노산 서열이다.
서열번호 31은 박테리아 세포에서의 발현을 위해서 사용되는 TIC868_10을 암호화하는 재조합 DNA 서열이다.
서열번호 32는 TIC868 변종인 TIC868_10을 암호화하는 식물 세포에서의 발현을 위한 합성 DNA 서열이다.
서열번호 33은 TIC868_10의 아미노산 서열이다.
서열번호 34는 박테리아 세포에서의 발현을 위해서 사용되는 TIC868_11을 암호화하는 재조합 DNA 서열이다.
서열번호 35는 식물 세포에서의 발현을 위한 TIC868_11을 암호화하는 합성 DNA 서열이다.
서열번호 36은 TIC868_11의 아미노산 서열이다.
서열번호 37은 박테리아 세포에서의 발현을 위해서 사용되는 TIC868_12를 암호화하는 재조합 DNA 서열이다.
서열번호 38은 식물 세포에서의 발현을 위한 TIC868_12를 암호화하는 합성 DNA 서열이다.
서열번호 39는 TIC868_12의 아미노산 서열이다.
서열번호 40은 식물 세포에서의 발현을 위한 TIC868_13을 암호화하는 합성 DNA 서열이다.
서열번호 41은 TIC868_13의 아미노산 서열이다.
서열번호 42는 식물 세포에서의 발현을 위한 TIC868_14를 암호화하는 합성 DNA 서열이다.
서열번호 43은 TIC868_14의 아미노산 서열이다.
서열번호 44는 식물 세포에서의 발현을 위한 TIC868_15를 암호화하는 합성 DNA 서열이다.
서열번호 45는 TIC868_15의 아미노산 서열이다.
서열번호 46은 식물 세포에서의 발현을 위한 TIC868_29를 암호화하는 합성 DNA 서열이다.
서열번호 47은 TIC868_29의 아미노산 서열이다.
서열번호 48은 박테리아 세포에서의 발현을 위해서 사용되는 TIC869를 암호화하는 재조합 DNA 서열이다.
서열번호 49는 식물 세포에서의 발현을 위한 TIC869를 암호화하는 합성 DNA 서열이다.
서열번호 50은 TIC869의 아미노산 서열이다.
서열번호 51은 박테리아 세포에서의 발현을 위해서 사용되는 TIC836을 암호화하는 재조합 DNA 서열이다.
서열번호 52는 식물 세포에서의 발현을 위한 TIC836을 암호화하는 합성 DNA 서열이다.
서열번호 53은 TIC836의 아미노산 서열이다.
농업 해충 방제 분야에서의 문제점은, 표적 해충에 대해서 효과적이고, 표적 해충 종에 대해서 넓은 스펙트럼의 독성을 나타내고, 바람직하지 않은 작물 문제를 유발하지 않으면서 식물에서 발현될 수 있고, 식물에서 상업적으로 사용되는 기존의 독소와 비교하여 대안적인 작용 기전을 제공하는 새로운 살곤충 단백질에 대한 요구를 특징으로 할 수 있다. 신규한 키메릭 살곤충 단백질이 본 명세서에서 개시되며, 그것은 특히 넓은 스펙트럼의 인시류 곤충 해충에 대해서 이러한 요구 각각을 충족시킨다.
현재 사용되는 살곤충 단백질에 대한 곤충 저항성의 발달을 회피하거나 또는 방지하기 위해서, 상이한 작용 기전(MOA), 뿐만 아니라 넓은 스펙트럼 및 효능을 갖는 새로운 살곤충 단백질이 인시류 제어를 위해서 필요하다. 이러한 요구를 해결하기 위한 한 방법은 상이한 생물 기원, 바람직하게는 박테리아, 진균 또는 식물로부터 새로운 살곤충 단백질을 개발하는 것이다. 또 다른 접근은 구조 유사성을 나타내는 다양한 Bt 단백질들 사이에서 분절을 교환하여 곤충 저해 특성을 갖는 새로운 키메릭 Bt 단백질을 생성하는 것이다. 당업계에 공지된 다수의 자연 존재 살곤충 결정 단백질의 도메인 구조를 재배열함으로써 향상된 특성을 갖는 키메릭 단백질을 생성할 가능성은 희박하다고 당업계에 공지되어 있다. 예를 들어, 문헌[A Strategy for Shuffling Numerous Bacillus thuringiensis Crystal Protein Domains." J. Economic Entomology, 97 (6) (2004): 1805-1813]을 참고하기 바란다.
본원에는 신규한 키메릭 살곤충 단백질을 암호화하는 재조합 핵산 분자 서열이 개시되어 있다. 이러한 살곤충 단백질은 개선된 살곤충 특성, 예컨대 더 넓은 스펙트럼의 표적 곤충 해충 종에 대해서 증가된 효능을 갖고, 상이한 작용 기전을 갖는 추가적인 독성 살곤충 단백질을 조작하려는 당업계의 계속적인 요구를 해결한다. 본 명세서에 개시된 예시적인 단백질을 비롯한, 이러한 군의 단백질의 구성원은 인시류 곤충 해충 종에 대해서 살곤충 활성을 나타낸다.
용어 "분절" 또는 "단편"는 개시된 키메릭 살곤충 단백질을 설명하는 완전한 아미노산 또는 핵산 서열보다 더 짧은 연속적인 아미노산 또는 핵산 서열을 기술하기 위해서 본 출원에서 사용된다. 곤충 저해 활성을 나타내는 분절 또는 단편가 또한 본 출원에 개시되어 있는데, 단 키메릭 살곤충 단백질의 상응하는 부분을 갖는 그러한 분절 또는 단편의 정렬이 분절 또는 단편와 키메릭 살곤충 단백질의 상응하는 부분 사이에서 약 65 내지 약 100%의 임의의 백분율의 아미노산 서열 동일성을 유발해야 한다.
용어 "활성인" 또는 "활성", "살충 활성" 또는 "살충" 또는 "살곤충 활성", "곤충 저해" 또는 "살곤충"에 대한 본 출원에서의 언급은 억제(성장, 섭취, 생식력, 또는 생존력의 억제), 억압(성장, 섭취, 생식력, 또는 생존력의 억압), 방제(해충 칩입 방제, 유효량의 살곤충 단백질을 함유하는 특정 작물에 대한 해충 섭취 활성의 제어) 또는 해충의 살상(해충의 이환(morbidity), 사멸, 또는 감소된 생식력을 유발함)에서의 독성제, 예컨대 살곤충 단백질의 효능을 말한다. 이러한 용어는 살충 유효량의 살곤충 단백질을 해충에 제공한 결과를 포함하고자 하며, 여기서 살곤충 단백질에 대한 해충의 노출은 이환, 사멸, 감소된 생식력, 또는 발육 저지를 유발한다. 이러한 용어는 또한 식물 내에 또는 식물 상에 살충 유효량의 살곤충 단백질을 제공한 결과로서, 식물, 식물 조직, 식물 부분, 종자, 식물 세포로부터의 해충의 퇴치, 또는 식물이 성장할 수 있는 특정 지리적인 위치로부터의 해충의 퇴치를 포함한다. 일반적으로, 살충 활성은 살충 단백질의 능력이 인시목 곤충을 포함하지만, 그에 제한되지 않는 특정 표적 해충의 이러한 단백질, 단백질 단편, 단백질 분절 또는 폴리뉴클레오타이드에 대한 곤충 섭식에 의해서 유발되는 성장, 발달, 생존력, 섭식 행동, 교배 행동 또는 생식력을 억제하거나, 부작용의 임의의 측정 가능한 감소를 억제하는 데 효과적인 것을 말한다. 살곤충 단백질은 식물에 의해서 생성될 수 있거나, 식물 또는 식물이 위치된 위치 내의 환경에 적용될 수 있다. 용어 "생물활성" "유효한" , "효과적인" 또는 그의 변형 용어가 또한 표적 곤충 해충에 대한 본 발명의 키메릭 살곤충 단백질의 효과를 기술하기 위해서 본 출원에서 상호 교환 가능하게 사용되는 용어이다.
표적 해충의 먹이에 제공되는 경우, 살충 유효량의 독성제는 독성제가 해충을 접촉할 때 살충 활성을 나타낸다. 독성제는 당업계에 공지된 살곤충 단백질제 또는 1종 이상의 화학 작용제일 수 있다. 살곤충 화학 작용제 및 살곤충 단백질제는 단독으로 또는 서로와 조합되어 사용될 수 있다. 화학 작용제에는 표적 해충의 억압을 위해서 특정 유전자를 표적화하는 dsRNA 분자, 유기염소, 유기인산염, 카르바메이트, 피레트로이드, 네오니코티노이드, 및 리아노이드가 포함되지만, 이들로 제한되는 것은 아니다. 살곤충 단백질제에는 본 출원에서 언급된 키메릭 살곤충 단백질, 뿐만 아니라 인시류 해충 종을 표적화하는 것을 비롯한 다른 단백질 독성제, 뿐만 아니라 다른 식물 해충을 방제하는 데 사용되는 단백질 독소, 예컨대 딱정벌레류, 총채벌레류, 노린재류 및 동시아류 종을 방제하는데 사용하기 위해서 당업계에서 입수 가능한 Cry 단백질이 포함된다.
해충, 특히 작물의 해충에 대한 언급은 작물의 곤충 해충, 특히 개시된 키메릭 살곤충 단백질에 의해서 방제되는 인시류 곤충 해충을 의미하고자 한다. 그러나, 해충에 대한 언급은 또한 딱정벌레류, 노린재류 및 동시아류의 식물 곤충 해충, 뿐만 아니라 선충 및 진균을 포함할 수 있는데, 이때 이러한 해충을 표적화하는 독성제는 키메릭 살곤충 단백질, 또는 키메릭 살곤충 단백질에 대해서 65 내지 약 100 동일한 단백질과 함께 국지화되거나 또는 함께 존재한다.
본 명세서에 개시된 키메릭 살곤충 단백질은 성충, 번데기, 애벌레, 및 유충을 비롯한 인시류 곤충 종, 뿐만 아니라 성충 및 유충을 비롯한 노린재 곤충 종으로부터의 곤충 해충에 대해서 살곤충 활성을 나타낸다. 인시목 곤충은 밤나방과(Family Noctuidae)의 멸강나방, 거세미 나방, 자나방, 및 헬리오틴(heliothine), 예를 들어 가을멸강충(스포돕테라 프루기페르다), 파밤나방(스포돕테라 엑시구아), 베르타 밤나방(마메스트라 콘피구라타), 검거세미 나방(아그로티스 입실론), 남방은무늬밤나방 애벌레(트리코플루시아 니), 대두 애벌레(수도플루시아 인클루덴스), 벨벳콩 자나방(안티카르시아 겜마탈리스), 그린 클로버웜(히페나 스카브라), 회색 담배 나방(헬리오티스 비레센스), 낟알 거세미 나방(아그로티스 수브테라네아), 멸강나방(수달레티아 유니펀크타), 서양 거세미 나방(아그로티스 오르토고니아); 명나방과로부터의 명나방(borer), 보호 고치를 만드는 유충(casebearer), 벌집 나방(webworm), 콘웜(coneworm), 캐비지웜(cabbageworm) 및 잎을 갉아먹는 인시류의 애벌레(skeletonizer), 예를 들어, 유럽 조명충 나방(오스트리니아 누빌랄리스), 네이블 오렌지 나방(아미엘로이스 트란시텔라), 옥수수 뿌리 벌집 나방(크람부스 칼리기노셀루스), 소드 벌집 나방(헤르페토그라마 리카르시살리스), 해바라기 나방(호모에오소마 엘렉텔룸), 명충 나방 애벌레(엘라스모팔푸스 리그노셀루스); 잎말이 나방과의 잎말이 나방(leafroller), 버드웜(budworm), 종자 벌레(seed worm), 및 과실 벌레(fruit worm), 예를 들어, 코들링 나방(시디아 포모넬라), 그레이프 베리 나방(엔도피자 비테아나), 복숭아순 나방(그라폴리타 몰레스타), 해바라기순 나방(술레이마 헬리안타나); 및 다수의 다른 경제적으로 중요한 인시류, 예를 들어, 배추좀 나방(플루텔라 크실로스텔라), 분홍 솜벌레(펙티노포라 고시피엘라) 및 매미 나방(리만트리아 디스파르)을 포함하지만 이들로 제한되는 것은 아니다. 인시목의 다른 곤충 해충은 예를 들어, 알라바마 아르길라세아(목화 잎 벌레), 아르킵스 아르기로스필라(과일 나무 잎말이 나방), 아르킵스 로사나(유럽 잎말이 나방) 및 다른 아르킵스 종, 킬로 서프레살리스(이화 명나방, 또는 쌀 명밤 나방), 크나팔로크로시스 메디날리스(혹명 나방), 크람부스 칼리기노셀루스(옥수수 뿌리 벌집 나방), 크람부스 테테렐루스(잔디 포충 나방), 디아트라에아 그란디오셀라(남서부 조명충 나방), 디아트라에아 사카랄리스(사탕수수 명나방), 에아리아스 인술라나(스피니 볼웜), 이아리아스 비텔라(스팟티드 볼웜), 헬리코베르파 아르미게라(미국 목화씨 벌레(American bollworm)), 헬리코베르파 제아(왕담배 밤나방 또는 목화씨 벌레), 헬리오티스 비레센스(회색 담배 나방), 헤르페토그라마 리카르시살리스(소드 벌집 나방), 로베시아 보트라나(유럽 포도나무 나방), 필록니스티스 시트렐라(귤굴 나방), 피에리스 브라시카에(큰 흰나비), 피에리스 라파에(배추 흰나비 앱러레, 또는 작은 흰나비), 플루텔라 크실로스텔라(배추좀 나방), 스포돕테라 엑시구아(파밤나방), 스포돕테라 리투라(담배 거세미 나방, 클러스터 캐터필라), 및 투타 앱솔루타(토마토 잎나방)를 포함한다.
본 출원에서 "단리된 DNA 분자", 또는 동등한 용어 또는 구에 대한 언급은 DNA 분자가 그의 본래 환경 내에 존재하지 않고, 단독으로 존재하거나 또는 다른 조성물과 조합되어 존재하는 것을 의미하고자 한다. 예를 들어, 유기체의 게놈의 DNA 내에서 본래 발견되는 핵산 요소, 예컨대 암호화 서열, 인트론 서열, 미번역 리더 서열, 프로모터 서열, 전사 중단 서열 등은, 그 요소가 유기체의 게놈 내에 존재하거나 그것이 본래 발견되는 게놈 내의 위치에 존재하는 한, "단리"되었다고 간주되지 않는다. 그러나, 이러한 요소 및 이러한 요소의 하위부분 각각은 그 요소가 유기체의 게놈 내에 그리고 그것이 본래 발견되는 게놈 내의 위치에 존재하지 않는 한 본 개시물의 범주 내에서 "단리"된 것이다. 유사하게, 살곤충 단백질 또는 그 단백질의 임의의 자연 발생 살곤충 변종을 암호화하는 뉴클레오타이드 서열은, 그 뉴클레오타이드 서열이 그 단백질을 암호화하는 서열이 본래 발견되는 박테리아의 DNA 내에 존재하지 않는 한 단리된 뉴클레오타이드 서열일 것이다. 자연 발생 살곤충 단백질의 아미노산 서열을 암호화하는 합성 뉴클레오타이드 서열은 본 개시물의 목적을 위해서 단리되었다고 간주될 것이다. 본 개시물의 목적을 위해서, 임의의 트랜스제닉 뉴클레오타이드 서열, 즉 식물 또는 박테리아의 세포의 게놈에 삽입되거나, 또는 염색체외 벡터에 존재하는 DNA의 뉴클레오타이드 서열은 그것이 세포를 형질전환시키기 위해서 사용된 플라스미드 또는 유사한 구조체 내에 존재하든, 식물 또는 박테리아의 게놈 내에 존재하든, 또는 식물 또는 박테리아로부터 유래된 조직, 자손, 생물학적 샘플 또는 상품에 검출가능한 양으로 존재하든 그렇지 않든 단리된 뉴클레오타이드 서열인 것으로 간주될 것이다.
실시예에서 추가로 기술된 바와 같이, 키메라지네시스(chimeragenesis) 노력을 통해서, 키메릭 살곤충 단백질을 암호화하는 대략 팔백사십사(844)개의 뉴클레오타이드 서열을 공지된 살곤충 독소의 프로톡신 및 독소 도메인(본 명세서에서 "모 단백질"이라 칭함)으로부터 구성하고, 발현시켜서, 인시류 활성에 대해서 생물학적 검정에서 시험하였다. 적은 수의 구성된 키메릭 살곤충 단백질이 그의 독소 성분이 유래된 모 단백질에 비해서 개선된 인시류 활성 또는 향상된 인시류 스펙트럼을 나타내었다.
개선된 인시류 활성 또는 향상된 인시류 스펙트럼을 갖는 이러한 신규한 키메릭 살곤충 단백질은 하기 살곤충 모 단백질 프로톡신 및 독소 도메인으로부터 구성되었다: Cry1Ah(도메인 I), Cry1Bb1(도메인 I 및 II), Cry 1Be2(도메인 I 및 II), Cry1Ja1(도메인 I 및 II), Cry1Fa1(도메인 I 및 II), Cry1Ac(도메인 II 및 프로톡신), Cry1Ca(도메인 III 및 프로톡신), Cry1Ka(도메인 III 및 프로톡신), Cry1Jx(도메인 III), Cry1Ab(도메인 III), Cry1Ab3(프로톡신), Cry1Da1(프로톡신), Cry4(프로톡신), Cry9(프로톡신), Cry1Be(프로톡신), 및 Cry1Ka(프로톡신).
구체적으로, 개선된 인시류 활성 또는 향상된 인시류 스펙트럼을 갖는 본 발명의 신규한 키메릭 살곤충 단백질은 하기 프로톡신 및 도메인 조합을 포함한다: TIC1100/서열번호 4(도메인 I- Cry1Ah, 도메인 II- Cry1Ac, 도메인 III- Cry1Ca, 프로톡신- Cry1Ac), TIC860/서열번호 7(도메인 I- Cry1Bb1, 도메인 II- Cry1BB1, 도메인 III- Cry1Ca, 프로톡신- Cry1Ac), TIC867/서열번호 10(도메인 I- Cry1Be2, 도메인 II- Cry1Be2, 도메인 III-Cry1Ka, 프로톡신- Cry1Ab3), TIC868/서열번호 28(도메인 I- Cry1Be2, 도메인 II-Cry1Be2, 및 도메인 III- Cry1Ca, 프로톡신- Cry1Ab3), TIC869/서열번호 50(도메인 I-Cry1Ja1, 도메인 II- Cry1Ja1, 도메인 III- Cry1Jx, 프로톡신-Cry1Ab3) 및 TIC836/서열번호 53(도메인 I-Cry1Fa1, 도메인 II-Cry1Fa1, 도메인 III- Cry1Ab, 프로톡신-Cry1Ac).
아미노산 치환 또는 대체 프로톡신 도메인이 도입된 변종이 또한 키메릭 살곤충 단백질 TIC867 및 TIC868을 위해서 구성될 수 있다. 구체적으로 TIC867 및 TIC868의 이러한 변종은 하기 아미노산 치환 및 대체 프로톡신 도메인을 포함할 수 있다: TIC867_20/서열번호 13(대체 프로톡신 도메인 Cry1Da1), TIC867_21/서열번호 16(대체 프로톡신 도메인 Cry4), TIC867_22/서열번호 19(대체 프로톡신 도메인 Cry9), TIC867_23/서열번호 21(대체 프로톡신 도메인 Cry1Be), TIC867_24/서열번호 23(대체 프로톡신 도메인 Cry1Ka), TIC867_25/서열번호 25(대체 프로톡신 도메인 Cry1Ka), TIC868_9/서열번호 30(아미노산 변형 N240S_Y343Q_N349T), TIC868_10/서열번호 33(대체 프로톡신 도메인 Cry1Da1), TIC868_11/서열번호 36(대체 프로톡신 도메인 Cry4), TIC868_12/서열번호 39(대체 프로톡신 도메인 Cry 9), TIC868_13/서열번호 41(대체 프로톡신 도메인 Cry1Be), TIC868_14/서열번호 43(대체 프로톡신 도메인 Cry1Ka), TIC868_15/서열번호 45(대체 프로톡신 도메인 Cry1Ca), 및 TIC868_29/서열번호 47(아미노산 변형 Q136Y_Y343Q_N349T).
실시예에 예증된 바와 같이, 이들 TIC867 및 TIC868 변종 각각은 인시류 활성을 변경하고/변경하거나 모 키메릭 살곤충 단백질의 인시류 활성 스펙트럼을 감소시켰는데, 따라서 이는 대체 프로톡신 도메인 및 아미노산 치환이 키메릭 살곤충 단백질 TIC867 및 TIC868의 살곤충 활성 및 스펙트럼에 직접적인 결과를 갖는다는 것을 나타낸다.
키메릭 살곤충 단백질 중 다수가 다양한 인시류 곤충 해충 종에 대해서 살곤충 활성을 나타낸다. 구체적으로, 본 출원에 개시된 신규한 키메릭 살곤충 단백질은 하기 인시류 곤충 해충 중 하나 이상에 대해서 활성을 나타내었다: 벨벳콩 자나방(VBC, 안티카르시아 겜마탈리스), 사탕수수 명나방(SCB, 디아트라에아 사카랄리스), 명충 나방 애벌레(LSCB, 엘라스모팔푸스 리그노셀루스), 왕담배 밤나방(CEW, 헬리코베르파 제아), 대두 팟웜(Soybean pod worm)(SPW, 헬리코베르파 제아), 목화씨 벌레(CBW, 헬리코베르파 제아), 회색 담배 나방(TBW, 헬리오티스 비레센스), 대두 애벌레(SBL, 크리소데익시스 인클루덴스), 블랙 멸강나방(Black armyworm)(BLAW, 스포돕테라 코스미오이데스), 남방 멸강나방(Southern armyworm)(SAW, 스포돕테라 에리다니아), 가을멸강충(FAW, 스포돕테라 프루기페르다), 파밤나방(BAW, 스포돕테라 엑시구아), 올드 월드 볼웜(OBW, 헬리코베르파 아르미게라), 오리엔탈 리프웜(Oriental leafworm)(OLW, 스포돕테라 리투라), 분홍 솜벌레(PBW, 펙티노포라 고시피엘라), 남서부 조명충 나방(SWCB, 디아트라에아 그란디오셀라), 스팟티드 볼웜(SBW, 에아리아스 비텔라), 미국 목화씨 벌레(American bollworm)(SABW, 헬리코베르파 겔로토페온), 및 해바라기 애벌레(Sunflower looper)(SFL, 라치플루시아 누). 따라서, 본 출원에 기술된 예시적인 단백질은 일반적인 기능과 관련되고, 성충, 애벌레 및 번데기를 비롯한 인시류 곤충 종으로부터의 곤충 해충에 대해서 살곤충 활성을 나타낸다.
당업계에 공지된 알고리즘을 기초로 다양한 컴퓨터를 사용하여 서로와 비교함으로써 키메릭 살곤충 단백질과 유사한 단백질을 식별할 수 있다. 예를 들어, 키메릭 살곤충 단백질에 관련된 단백질의 아미노산 서열 동일성은 다음 디폴트 파라미터를 사용하여 클러스탈(Clustal) W 정렬을 사용하여 분석될 수 있다: 가중치 매트릭스: 블로섬(blosum), 갭 오프닝 페널티(Gap opening penalty): 10.0, 갭 익스텐션 페널티(Gap extension penalty): 0.05, 친수성 갭(Hydrophilic gap): 온(On), 친수성 잔기(Hydrophilic residue): GPSNDQERK, 잔기-특이적 갭 페널티(Residue-specific gap penalty): 온(문헌 [Thompson, et al (1994) Nucleic Acids Research, 22:4673-4680]). (아미노산 동일성/해당 단백질의 길이) x 100%의 값에 의해서 아미노산 동일성 백분율을 추가로 계산한다. 다른 정렬 알고리즘이 또한 당업계에서 사용 가능하고, 클러스탈 W 정렬을 사용하여 수득된 것과 유사한 결과를 제공하고, 본 출원에서 고려된다.
곤충 저해 활성을 나타내는 쿼리(query) 단백질이 본 출원에서 개시되며, 단 대상 키메릭 살곤충 단백질을 갖는 그러한 쿼리 단백질의 정렬은 서열번호 4, 7, 10, 13, 16, 19, 21, 23, 25, 28, 30, 33, 36, 39, 41, 43, 45, 47, 50 및 53에 제시되어 있고, 쿼리 단백질과 대상 단백질 간의 아미노산 서열 동일성은 적어도 약 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 또는 약 100% (또는 이 범위의 임의의 백분율)이어야 한다.
본 출원의 실시예에서 추가로 기술된 바와 같이, 키메릭 살곤충 단백질을 암호화하는 합성 또는 인공 서열을 식물에서 사용하기 위해서 설계하였다. 식물에서 사용하기 위해서 설계된 예시적인 합성 뉴클레오타이드 서열은 서열번호 2 및 3(TIC1100), 서열번호 6(TIC860), 서열번호 9(TIC867), 서열번호 12(TIC867_20), 서열번호 15(TIC867_21), 서열번호 18(TIC867_22), 서열번호 20(TIC867_23), 서열번호 22(TIC867_24), 서열번호 24(TIC867_25), 서열번호 27(TIC868), 서열번호 29(TIC868_9), 서열번호 32(TIC868_10), 서열번호 35(TIC868_11), 서열번호 38(TIC868_12), 서열번호 40(TIC868_13), 서열번호 42(TIC868_14), 서열번호 44(TIC868_15), 서열번호 46(TIC868_29), 서열번호 49(TIC869) 및 서열번호 52(TIC836)에 명시된다.
식물 세포에서의 발현을 위해서, 키메릭 살곤충 단백질은 발현되어 시토졸 내에 잔류할 수 있거나 식물 세포의 다양한 세포 소기관으로 표적화될 수 있다. 예를 들어, 단백질의 엽록체로의 표적화은 오프-표현형(off-phenotype)이 발생하는 것을 방지하면서 트랜스제닉 식물에서 발현되는 단백질의 수준을 증가시킬 수 있다. 표적화는 또한 유전자 이식 이벤트에서 해충 저항성 효능을 증가시킬 수 있다. 표적 펩타이드 또는 트랜지트(transit) 펩타이드는 핵, 미토콘드리아, 소포체(ER), 엽록체, 아포플라스트, 퍼옥시좀 및 원형질 막을 비롯한 세포의 특정 구역으로의 단백질의 이송을 유도하는 짧은(3 내지 70개의 아미노산 길이) 펩타이드 쇄이다. 일부 타겟 펩타이드는 단백질이 이송된 후 신호 펩티다제에 의해서 단백질로부터 절단된다. 엽록체로의 표적화를 위해서, 단백질은 대략 40 내지 50개의 아미노산인 트랜지트 펩타이드를 함유한다. 엽록체 트랜지트 펩타이드의 사용의 설명에 대해서는, 미국 특허 제5,188,642호 및 제5,728,925호를 참고하기 바란다. 많은 엽록체-국지화된 단백질은 전구체로서 핵 유전자로부터 발현되어, 엽록체 트랜지트 펩타이드(CTP)에 의해서 엽록체로 표적화된다. 그러한 단리된 엽록체 단백질의 예에는 리불로오스-1,5,-비스포스페이트 카르복실라제, 페레독신, 페레독신 옥시도리덕타제, 광수확 복합 단백질(light-harvesting complex protein) I 및 단백질 II, 티오레독신 F, 엔올피루빌 시키메이트 포스페이트 신타제(enolpyruvyl shikimate phosphate synthase)(EPSPS), 및 미국 특허 제7,193,133호에 기술된 트랜지트 펩타이드의 소단위체(SSU)와 연관된 것이 포함되지만, 이들로 제한되는 것은 아니다. 비-엽록체 단백질이 이종 CTP를 갖는 단백질 융합의 사용에 의해서 엽록체로 표적화될 수 있고, 그 CTP는 엽록체로 단백질을 표적화하기에 충분하다는 것이 생체내 및 생체외 실험에서 예증되어 있다. 적합한 엽록체 트래지트 펩타이드, 예컨대 아라비돕시스 탈리아나(Arabidopsis thaliana) EPSPS CTP(CTP2)(문헌 [Klee et al., Mol. Gen. Genet. 210:437-442, 1987] 참고) 또는 페츄니아 히브리다(Petunia hybrida) EPSPS CTP(CTP4)(문헌 [della-Cioppa et al., Proc. Natl. Acad. Sci. USA 83:6873-6877, 1986] 참고)의 도입이 트랜스제닉 식물에서 이종 EPSPS 단백질 서열을 엽록체로 표적화하는 것이 밝혀져 있다(미국 특허 제5,627,061호; 제5,633,435호; 및 제5,312,910호; 및 유럽 특허 제EP 0218571호; 제EP 189707호; 제EP 508909호; 및 제EP 924299호 참고). 키메릭 살곤충 단백질의 엽록체로의 표적화를 위해서, 엽록체 트랜지트 펩타이드를 암호화하는 서열은 식물 세포에서 최적의 발현을 위해서 설계된 키메릭 살곤충 단백질을 암호화하는 합성 암호화 서열에 작동가능한 연결 및 프레임으로 5' 자리에 위치된다.
당업계에 공지된 형질전환 방법 및 기술에 따라서 이러한 합성 또는 인공 뉴클레오타이드 서열을 함유하는 발현 카세트 및 벡터를 구성하였고, 옥수수, 목화 및 대두 식물 세포에 도입하였다. 형질전환된 세포(transformed cell)를 키메릭 살곤충 단백질을 발현하는 것으로 관찰된 형질전환된 식물 내에서 재생시켰다. 살충 활성을 시험하기 위하여, 형질전환된 식물로부터 수득된 식물 잎 디스크를 사용하여 인시류 해충 유충의 존재 하에서 생물학적 검정을 수행하였다. 키메릭 살곤충 단백질을 암호화하는 재조합 핵산 분자 조성물이 고려된다. 예를 들어, 키메릭 살곤충 단백질은 재조합 DNA 구조체를 사용하여 발현될 수 있는데, 그 구조체에서 키메릭 살곤충 단백질을 암호화하는 ORF를 갖는 폴리뉴클레오타이드 분자는 유전자 발현 요소, 예컨대 프로모터, 및 구조체가 의도되는 시스템에서 발현을 위해서 필요한 임의의 다른 조절 요소에 작동 가능하게 연결되어 있다. 비제한적인 예에는 식물에서의 키메릭 살곤충 단백질의 발현을 위한 합성 키메릭 살곤충 단백질 암호화 서열에 작동 가능하게 연결된 식물-기능성 프로모터 또는 Bt 박테리아 또는 다른 바실러스 종에서의 단백질의 발현을 위한 키메릭 살곤충 단백질 암호화 서열에 작동 가능하게 연결된 Bt-기능성 프로모터가 포함된다. 인핸서(enhancer), 인트론, 미번역 리더, 암호화된 단백질 부동화 태그(immobilization tag)(HIS-tag), 위치 이동 펩타이드(즉, 색소체 트랜지트 펩타이드, 신호 펩타이드), 번역 후 변형 효소(post-translational modifying enzyme)를 위한 폴리펩타이드 서열, 리보솜 결합 자리, 및 RNAi 타겟 자리를 포함하지만, 그에 제한되지 않는 다른 요소가 키메릭 살곤충 단백질 암호화 서열에 작동 가능하게 연결될 수 있다.
본 명세서에 제공된 예시적인 재조합 폴리뉴클레오타이드 분자에는 서열번호 4(TIC1100), 7(TIC860), 10(TIC867), 13(TIC867_20), 16(TIC867_21), 19(TIC867_22), 21(TIC867_23), 23(TIC867_24), 25(TIC867_25), 28(TIC868), 30(TIC868_9), 33(TIC868_10), 36(TIC868_11), 39(TIC867_12), 41(TIC867_13), 43(TIC867_14), 45(TIC867_15), 47(TIC867_29), 50(TIC869) 및 53(TIC836)에 명시된 바와 같은 아미노산 서열을 갖는 폴리펩타이드 또는 단백질을 암호화하는 폴리뉴클레오타이드, 예컨대 서열번호 1, 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 22, 24, 26, 27, 29, 31, 32, 34, 35, 37, 38, 40, 42, 44, 46, 48, 49, 51, 및 52에 작동 가능하게 연결된 이종 프로모터가 포함되지만, 이들로 제한되는 것은 아니다. 이종 프로모터는 또한 색소체 표적화된 키메릭 살곤충 단백질 및 표적화되지 않은 키메릭 살곤충 단백질을 암호화하는 합성 DNA 암호화 서열에 작동 가능하게 연결될 수 있다. 본 명세서에 개시된 키메릭 살곤충 단백질을 암호화하는 재조합 핵산 분자의 코돈은 동의 코돈(동의 치환(silent substitution)으로 당업계에서 공지됨)에 의해서 치환될 수 있다고 고려된다.
키메릭 살곤충 단백질 암호화 서열을 포함하는 재조합 DNA 분자 또는 구조체는 키메릭 살곤충 단백질을 암호화하는 DNA 서열, 키메릭 살곤충 단백질과 상이한 단백질, 곤충 저해 dsRNA 분자, 또는 보조적 단백질에 부수적으로 또는 그들과 함께 발현하도록 구성될 수 있는 하나 이상의 독성제를 암호화하는 DNA 구역을 추가로 포함할 수 있다. 보조 단백질은, 예를 들어 발현의 도움, 식물에서의 안정성에 영향을 미침, 올리고머화를 위한 자유에너지를 최적화함, 독성을 증가시킴, 그리고 활성 범위를 증가시킴으로써, 보조인자, 효소, 결합-파트너, 또는 곤충 저해 제제의 효율성을 돕는 기능을 하는 다른 제제를 포함하지만, 이들로 제한되지는 않는다. 보조 단백질은, 예를 들어 하나 이상의 곤충 저해 제제의 흡수를 용이하게 하거나, 또는 독성 제제의 독성 효과를 강화시킬 수 있다.
재조합 DNA 분자 또는 구조체는 모든 단백질 또는 dsRNA 분자가 하나의 프로모터로부터 발현되거나 각각의 단백질 또는 dsRNA 분자가 개별 프로모터 제어 또는 그의 일부 조합 하에 있도록 조립될 수 있다. 본 발명의 단백질은 키메릭 살곤충 단백질이 선택된 발현 시스템의 유형의 따라서, 다른 개방 판독 프레임(open reading frame) 및 프로모터를 또한 함유하는 일반적인 뉴클레오타이드 분절로부터 발현되는 다중-유전자 발현 시스템으로부터 발현될 수 있다. 예를 들어, 박테리아 다중-유전자 발현 시스템은 단일 프로모터를 사용하여 단일 오페론(operon) 내로부터의 다중-연결/탠덤(tandem) 개방 판독 프레임의 발현(즉, 다시스트론 발현(polycistronic expression))을 유도할 수 있다. 또 다른 예에서, 식물 다중-유전자 발현 시스템은 상이한 단백질 또는 다른 독성제, 예컨대 1종 이상의 dsRNA 분자를 각각 발현하는 다중-미연결 발현 카세트를 사용할 수 있다.
키메릭 살곤충 단백질 암호화 서열을 포함하는 재조합 핵산 분자 또는 재조합 DNA 구조체는 벡터, 예를 들어, 플라스미드, 바큘로바이러스, 합성 염색체, 비리온, 코스미드, 파지미드, 파지(phage), 바이러스 벡터에 의해서 숙주 세포로 전달될 수 있다. 그러한 벡터를 사용하여 숙주 세포에서 키메릭 살곤충 단백질 암호화 서열의 안정한 발현 또는 일시적인 발현을 성취할 수 있거나, 암호화된 폴리펩타이드의 후속 발현을 성취할 수 있다. 키메릭 살곤충 단백질 서열 암호화 서열을 포함하고, 숙주 세포에 도입되는 외인성 재조합 폴리뉴클레오타이드 또는 재조합 DNA 구조체를 본 명세서에서 "이식 유전자"라 칭한다.
키메릭 살곤충 단백질 중 임의의 1종 이상을 암호화하는 폴리뉴클레오타이드를 함유하는 트랜스제닉 박테리아, 트랜스제닉 식물 세포, 트랜스제닉 식물, 및 트랜스제닉 식물 부분이 본 명세서에서 제공된다. 용어 "박테리아 세포" 또는 "박테리아"에는 아그로박테리움, 바실러스, 에쉐리키아, 살로넬라, 슈도모나스, 또는 리조븀 세포가 포함되지만, 이들로 제한되는 것은 아니다. 용어 "식물 세포" 또는 "식물"에는 쌍떡잎 식물 세포 또는 외떡잎 식물 세포가 포함될 수 있지만, 이들로 제한되는 것은 아니다. 고려되는 식물 및 식물 세포에는 자주개자리, 바나나, 보리, 콩, 브로콜리, 양배추, 배추속 식물, 당근, 카사바, 카스터(castor), 콜리플라워, 셀러리, 병아리콩, 배추, 감귤, 코코넛, 커피, 옥수수, 토끼풀, 목화, 조롱박, 오이, 미송, 가지, 유칼립투스, 아마, 마늘, 포도, 홉, 리크(leek), 상추, 로브롤리 소나무(Loblolly pine), 수수, 멜론, 견과, 귀리, 올리브, 양파, 풍치림, 야자, 목초, 완두, 땅콩, 후추, 나무콩, 소나무, 감자, 포플러, 호박, 라디에타 파인, 유채, 벼, 루트스톡(rootstock), 호밀, 홍화, 관목, 수수, 서던 파인(Southern pine), 대두, 시금치, 스쿼시(squash), 딸기, 사탕무, 사탕수수, 해바라기, 스위트 콘, 스위트 검(sweet gum), 고구마, 스위치그래스(switchgrass), 차, 담배, 토마토, 트리티케일, 잔디풀, 수박, 및 밀 식물 세포 또는 식물이 포함되지만, 이들로 제한되는 것은 아니다. 특정 실시형태에서, 트랜스제닉 식물 세포로부터 재생된 트랜스제닉 식물 및 트랜스제닉 식물 부분이 제공된다. 특정 실시형태에서, 트랜스제닉 식물은 절단, 스냅핑(snapping), 그라인딩 또는 식물로부터 부분을 달리 자르는 것에 의해서 트랜스제닉 종자로부터 수득될 수 있다. 특정 실시형태에서, 식물 부분은 종자, 꼬투리(boll), 잎, 꽃, 줄기, 뿌리 또는 그의 임의의 일부, 또는 트랜스제닉 식물 부분의 재생불가능한 일부일 수 있다. 본 명세서에서 사용되는 바와 같이, 트랜스제닉 식물 부분의 "재생불가능한 일부"는 전체 식물을 형성하도록 유도될 수 없는 일부 또는 유성 생식 및/또는 무성 생식할 수 있는 전체 식물을 형성하도록 유도될 수 없는 일부이다. 특정 실시형태에서, 식물 부분의 재생불가능한 일부는 트랜스제닉 종자, 꼬투리, 잎, 꽃, 줄기 또는 뿌리의 일부이다.
인시류-억제량의 키메릭 살곤충 단백질을 포함하는 트랜스제닉 식물의 생산 방법이 제공된다. 그러한 식물은 본 출원에 제공된 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드를 식물 세포에 도입하고, 곤충 또는 인시류-억제량의 키메릭 살곤충 단백질을 발현하는 상기 식물 세포로부터 유래된 식물을 선별함으로써 생산될 수 있다. 식물은 재생, 종자, 화분, 또는 분열 조직 형질전환 기술에 의해서 식물 세포로부터 유래될 수 있다. 식물의 형질전환 방법은 당업계에 공지되어 있다. 예를 들어, 아그로박테리움-매개된 형질전환은 미국 특허 출원 공개 제2009/0138985A1호(대두), 제2008/0280361A1호(대두), 제2009/0142837A1호(옥수수), 제2008/0282432호(목화), 및 제2008/0256667호(목화)에 기술되어 있다.
키메릭 살곤충 단백질을 발현하는 식물은 다른 살곤충 단백질을 발현하는 유전자 이식 이벤트와 함께 성장하고/성장하거나 트랜스제닉 특질, 예컨대 다른 곤충 제어 특질, 제초제 내성 유전자, 유전자 부여 수율(conferring yield) 또는 스트레스 내성 특질 등을 발현시킴으로써 이종 교배될 수 있거나, 또는 그러한 특질은 특질이 모두 연결되도록 단일 벡터 내에 조합될 수 있다.
검출가능한 양의 키메릭 살곤충 단백질, 그의 곤충 저해 분절 또는 단편, 또는 그의 임의의 구별되는 부분을 포함하는 가공된 식물 생성물이 또한 본 출원에서 개시된다. 특정 실시형태에서, 가공된 생성물은 식물 부분, 식물 바이오매스, 오일, 으깬 곡물(meal), 당, 동물 사료, 곡물 가루, 플레이크, 겨, 린트, 깍지, 가공된 종자, 및 종자로 이루어진 군으로부터 선택된다. 특정 실시형태에서, 가공된 생성물은 재생불가능하다. 식물 생성물은 트랜스제닉 식물 또는 트랜스제닉 식물 부분으로부터 유래된 상품 또는 다른 상업 제품을 포함할 수 있고, 여기서 상품 또는 다른 제품은 키메릭 살곤충 단백질의 구별되는 부분을 암호화하거나 포함하는 뉴클레오타이드 분절 또는 발현된 RNA 또는 단백질을 검출함으로써 상거래를 통해서 추적될 수 있다.
키메릭 살곤충 단백질을 사용하여 작물의 곤충, 특히 인시류 침입을 방제하는 방법이 또한 본 출원에서 개시된다. 그러한 방법은 곤충-억제량 또는 인시류-억제량의 키메릭 살곤충 단백질을 포함하는 식물을 성장시키는 단계를 포함한다. 특정 실시형태에서, 그러한 방법은 (i) 식물 또는 식물을 발생시키는 종자에 키메릭 살곤충 단백질을 포함하거나 암호화하는 임의의 조성물을 적용하는 것; 및 (ii) 식물 또는 식물을 발생시키는 식물 세포를 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드로 형질전환시키는 것 중 임의의 하나 이상을 추가로 포함할 수 있다. 일반적으로, 키메릭 살곤충 단백질이 조성물, 미생물 또는 트랜스제닉 식물에 제공되어 인시류 곤충에 대한 곤충 저해 활성을 부여할 수 있다고 고려된다.
특정 실시형태에서, 키메릭 살곤충 단백질은 재조합 바실러스 또는 발현에 적합한 조건 하에서 키메릭 살곤충 단백질을 발현하도록 형질전환된 임의의 다른 재조합 박테리아 세포를 배양함으로써 제조된 곤충 저해 조성물의 살곤충 활성 성분이다. 그러한 조성물은 키메릭 살곤충 단백질을 발현/생성하는 그러한 재조합 세포의 배양액의 건조, 동결 건조, 균질화, 추출, 여과, 원심 분리, 침강, 또는 농축에 의해서 제조될 수 있다. 그러한 방법은 바실러스 또는 다른 곤충병원성 박테리아 세포 추출물, 세포 현탁액, 세포 분쇄액, 세포 용해물, 세포 상청액, 세포 여과물, 또는 세포 펠렛을 생성할 수 있다. 이렇게 제조된 키메릭 살곤충 단백질을 수득함으로써, 키메릭 살곤충 단백질을 포함하는 조성물은 박테리아 세포, 박테리아 포자, 및 부아포 봉입체(parasporal inclusion body)를 포함할 수 있고, 농업용 곤충 저해 분무 제품으로서 또는 섭식 생물학적 검정에서의 곤충 저해 제제로서 다양한 용도를 위해서 제제화될 수 있다.
상기에 언급된 화합물 또는 제제는 농업용으로 허용가능한 담체, 예컨대 미끼, 분말, 더스트, 펠렛, 과립, 스프레이, 에멀젼, 콜로이드 현탁액, 수용액, 바실러스 포자 또는 결정 제제 또는 종자 처리제를 추가로 포함할 수 있다. 그러한 화합물 또는 제제는 또한 단백질 중 1종 이상을 발현하도록 형질전환된 재조합 식물 세포, 식물 조직, 종자 또는 식물; 또는 단백질 중 1종 이상을 발현하도록 형질전환된 박테리아를 추가로 포함할 수 있다. 재조합 폴리펩타이드에 내재하는 곤충 활성 또는 살곤충 저해 수준 및 식물 또는 섭식 검정에 적용될 화합물 또는 제제의 수준에 따라서, 화합물 또는 제제는 다양한 중량% 양의 재조합 폴리펩타이드, 예를 들어 0.0001 중량% 내지 0.001 중량% 내지 0.01 중량% 내지 1% 내지 99 중량%의 재조합 폴리펩타이드를 포함할 수 있다.
실시형태에서, 저항성 발전 가능성을 감소시키기 위해서, 키메릭 살곤충 단백질을 포함하는 곤충 저해 조성물 또는 트랜스제닉 식물은 동일한 인시류 곤충 종에 대해서 곤충 저해 활성을 나타내지만, 키메릭 살곤충 단백질과 상이한 적어도 1종의 추가의 독성제를 추가로 포함할 수 있다. 그러한 조성물을 위해서 가능한 추가의 독성제는 곤충 저해 단백질 및 곤충 저해 dsRNA 분자를 포함한다. 곤충 해충을 방제하기 위해서 그러한 리보뉴클레오타이드 서열을 사용한 한 예가 바움(Baum) 등의 특허(미국 특허 공개 제2006/0021087 A1호)에 기술되어 있다. 인시류 해충의 방제를 위한 그러한 추가 폴리펩타이드(들)는 곤충 저해 단백질, 예컨대 Cry1A(미국 특허 제5,880,275호), Cry1Ab, Cry1Ac, Cry1A.105, Cry1Ae, Cry1B(미국 특허 공개 제10/525,318호), Cry1C(미국 특허 제6,033,874호), Cry1D, Cry1E, Cry1F 및 Cry1A/F 키메라(미국 특허 제7,070,982호; 제6,962,705호; 및 제6,713,063호), Cry1G, Cry1H, Cry1I, Cry1J, Cry1K, Cry1L, Cry2A, Cry2Ab(미국 특허 제7,064,249호), Cry2Ae, Cry4B, Cry6, Cry7, Cry8, Cry9, Cry15, Cry43A, Cry43B, Cry51Aa1, ET66, TIC400, TIC800, TIC834, TIC1415, Vip3A, VIP3Ab, VIP3B, AXMI-001, AXMI-002, AXMI-030, AXMI-035, 및 AXMI-045(미국 특허 공개 제2013-0117884 A1호), AXMI-52, AXMI-58, AXMI-88, AXMI-97, AXMI-102, AXMI-112, AXMI-117, AXMI-100(미국 특허 공개 제2013-0310543 A1호), AXMI-115, AXMI-113, AXMI-005(미국 특허 공개 제2013-0104259 A1호), AXMI-134(미국 특허 공개 제2013-0167264 A1호), AXMI-150(미국 특허 공개 제2010-0160231 A1호), AXMI-184(미국 특허 공개 제2010-0004176 A1호), AXMI-196, AXMI-204, AXMI-207, AXMI-209(미국 특허 공개 제2011-0030096 A1호), AXMI-218, AXMI-220(미국 특허 공개 제2014-0245491 A1호), AXMI-221z, AXMI-222z, AXMI-223z, AXMI-224z, AXMI-225z(미국 특허 공개 제2014-0196175 A1호), AXMI-238(미국 특허 공개 제2014-0033363 A1호), AXMI-270(미국 특허 공개 제2014-0223598 A1호), AXMI-345(미국 특허 공개 제2014-0373195 A1호), DIG-3(미국 특허 공개 제2013-0219570 A1호), DIG-5(미국 특허 공개 제2010-0317569 A1호), DIG-11(미국 특허 공개 제2010-0319093 A1호), AfIP-1A 및 그의 유도체(미국 특허 공개 제2014-0033361 A1호), AfIP-1B 및 그의 유도체(미국 특허 공개 제2014-0033361 A1호), PIP-1APIP-1B(미국 특허 공개 제2014-0007292 A1호), PSEEN3174(미국 특허 공개 제2014-0007292 A1호), AECFG-592740(미국 특허 공개 제2014-0007292 A1호), Pput_1063(미국 특허 공개 제2014-0007292 A1호), Pput_1064(미국 특허 공개 제2014-0007292 A1호), GS-135 및 그의 유도체(미국 특허 공개 제2012-0233726 A1호), GS153 및 그의 유도체(미국 특허 공개 제2012-0192310 A1호), GS154 및 그의 유도체(미국 특허 공개 제2012-0192310 A1호), GS155 및 그의 유도체(미국 특허 공개 제2012-0192310 A1호), 미국 특허 공개 제2012-0167259 A1호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2012-0047606 A1에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2011-0154536 A1호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2011-0112013 A1호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2010-0192256 A1호에 기술된 바와 같은 서열번호 2 및 4 및 그의 유도체, 미국 특허 공개 제2010-0077507 A1호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2010-0077508 A1호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2009-0313721 A1호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 공개 제2010-0269221 A1호에 기술된 바와 같은 서열번호 2 또는 4 및 그의 유도체, 미국 특허 제7,772,465 (B2)호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 국제 특허 제WO2014/008054 A2호에 기술된 바와 같은 CF161_0085 및 그의 유도체, 미국 특허 공개 제US2008-0172762 A1호, 제US2011-0055968 A1호, 및 제US2012-0117690 A1호에 기술된 바와 같은 인시류 독소 단백질 및 그의 유도체; 미국 특허 제US7510878(B2)호에 기술된 바와 같은 서열번호 2 및 그의 유도체, 미국 특허 제7812129(B1)호에 기술된 바와 같은 서열번호 2 및 그의 유도체 등(이들로 제한되지 않음)으로 이루어진 군으로부터 선택될 수 있다.
다른 실시형태에서, 곤충 저해 조성물 또는 트랜스제닉 식물은 수득된 곤충 저해 스펙트럼을 확장시키기 위해서, 본 발명의 키메릭 살곤충 단백질에 의해서 억제되지 않는 곤충 해충(예컨대, 딱정벌레류, 노린재류 및 동시아류 해충)에 대해서 곤충 저해 활성을 나타내는 적어도 1종의 추가 독성제를 추가로 포함할 수 있다.
딱정벌레류 해충의 방제를 위한 그러한 추가 독성제는 곤충 저해 단백질, 예컨대 Cry3Bb(미국 특허 제6,501,009호), Cry1C 변종, Cry3A 변종, Cry3, Cry3B, Cry34/35, 5307, AXMI134(미국 특허 공개 제2013-0167264 A1호), AXMI-184(미국 특허 공개 제2010-0004176 A1호), AXMI-205(미국 특허 공개 제2014-0298538 A1호), axmi207(미국 특허 공개 제2013-0303440 A1호), AXMI-218, AXMI-220(미국 특허 공개 제20140245491A1호), AXMI-221z, AXMI-223z(미국 특허 공개 제2014-0196175 A1호), AXMI-279(미국 특허 공개 제2014-0223599 A1호), AXMI-R1 및 그의 변종(미국 특허 공개 제2010-0197592 A1호), TIC407, TIC417, TIC431, TIC807, TIC853, TIC901, TIC1201, TIC3131, DIG-10(미국 특허 공개 제2010-0319092 A1호), eHIPs(미국 특허 출원 공개 제2010/0017914호), IP3 및 그의 변종(미국 특허 공개 제2012-0210462 A1호), 및 ω-헥사톡신-Hv1a(미국 특허 출원 공개 제US2014-0366227 A1호) (이에 제한되지 않음)로 이루어진 군으로부터 선택될 수 있다.
노린재류 해충의 방제를 위한 그러한 추가 독성제는 노린재류 활성 단백질, 예컨대 TIC1415(미국 특허 공개 제2013-0097735 A1호), TIC807(미국 특허 제8609936호), TIC834(미국 특허 공개 제2013-0269060 A1호), AXMI-036(미국 특허 공개 제2010-0137216 A1호), 및 AXMI-171(미국 특허 공개 제2013-0055469 A1호) (이에 제한되지 않음)로 이루어진 군으로부터 선택될 수 있다. 딱정벌레류, 인시류, 및 노린재류 곤충 해충의 방제를 위한 추가 폴리펩타이드는 네일 크릭모어(Neil Crickmore)에 의해서 유지되는 바실러스 투린기엔시스 독소 명명법 웹사이트(www.btnomenclature.info)에서 찾아볼 수 있다.
키메릭 살곤충 단백질-암호화 서열 및 키메릭 살곤충 단백질에 대해서 상당한 백분율의 동일성을 갖는 서열은 당업자에게 공지된 방법, 예를 들어 중합효소 연쇄 반응(PCR), 열 증폭 및 혼성화를 사용하여 식별될 수 있다. 예를 들어, 키메릭 살곤충 단백질을 사용하여 관련 단백질에 특이적으로 결합하는 항체를 생성할 수 있고, 그것을 사용하여 스크리닝하여 밀접하게 관련된 다른 단백질을 찾을 수 있다.
추가로, 키메릭 살곤충 단백질을 암호화하는 뉴클레오타이드 서열을 스크리닝용 프로브 또는 프라이머로서 사용하여 열 사이클 또는 등온 증폭 및 혼성화 방법을 사용하여 그러한 부류의 다른 구성원을 식별할 수 있다. 예를 들어, 서열번호 2에 나타낸 바와 같은 서열로부터 유래된 올리고뉴클레오타이드를 사용하여, 상품으로부터 유래된 데옥시리보핵산 샘플 내의 키메릭 살곤충 이식 유전자 유무를 측정할 수 있다. 올리고뉴클레오타이드를 사용한 특정 핵산 검출 방법의 민감성을 고려할 때, 서열번호 2 중 어느 하나에 명시된 바와 같은 서열로부터 유래된 올리고뉴클레오타이드를 사용하여 상품의 일부만이 서열번호 2 중 임의의 것을 함유하는 트랜스제닉 식물로부터 유래된 풀드 소스(pooled source)로부터 유래된 상품 내에서 각각의 키메릭 살곤충 단백질을 검출할 수 있다고 예상된다.
실시예
상기 내용에 비추어, 당업자는 하기에 개시된 실시형태가 본 발명을 단지 대표하는 것이며, 이것은 다양한 형태로 포함될 수 있다는 것을 인지할 것이다. 따라서, 본 명세서에 개시된 구체적인 구조적 상세사항 및 기능적 상세사항은 제한으로서 이해되어서는 안 된다.
실시예 1
인시류-활성인 신규한 키메릭 살곤충 단백질 암호화 서열의 생성 및 클로닝
본 실시예는 신규한 키메릭 살곤충 단백질의 생성 및 키메릭 살곤충 단백질의 클로닝 및 발현을 설명한다.
공지된 Cry 단백질 유전자로부터 재조합 핵산 서열을 구성하여 신규한 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드 서열을 생성하였다. 생성된 폴리뉴클레오타이드 서열을 바실러스 투린기엔시스(Bt) 발현 플라스미드 벡터에 클로닝하였다. 폴리뉴클레오타이드 서열의 확인 후, 발현 플라스미드를 Bt에 형질전환시키고, 발현시켰다. 발현된 신규한 키메릭 단백질의 제제를 다양한 인시류 해충에 대한 활성에 대해서 검정하였다.
키메릭 살곤충 단백질을 암호화하는 다수의 폴리뉴클레오타이드 서열을 생성하였고, 생물학적 검정에서 시험하였다. 키메릭 살곤충 단백질 전부가 활성을 나타낸 것은 아니었다. 생물학적 검정에서 예증된 특정 인시류에 대한 활성을 기초로 단지 몇몇의 키메릭 살곤충 단백질을 선별하였다. 본래 키메릭 살곤충 단백질 TIC867 및 TIC868을 기반으로 하는 아미노산 치환 또는 대체 프로톡신 도메인이 도입된 아미노산 변종을 또한 생성하였다. 본 발명의 키메릭 살곤충 단백질(도메인 I, II 및 III, 프로톡신)의 성분을 표 1에 나타낸다. 본래 TIC868 단백질 서열에 대한 TIC868 변종에서의 아미노산 치환을 또한 나타낸다.
실시예 2
신규한 키메릭 살곤충 단백질은 인시류 해충에 대해서 활성을 나타냄
본 실시예는 실시예 1에 기술된 키메릭 살곤충 단백질의 시험 및 키메릭 살곤충 단백질에 대해서 관찰된 인시류 활성을 설명한다.
키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드 서열을 Bt에서 발현시켰다. 이어서, 발현된 키메릭 살곤충 단백질을 옥수수, 사탕수수, 대두, 및 목화, 뿐만 아니라 다른 작물의 해충인 것으로 공지된 다양한 인시류에 대해서 검정하였다. 구체적으로, 살곤충 단백질을 벨벳콩 자나방(VBC, 안티카르시아 겜마탈리스), 사탕수수 명나방(SCB, 디아트라에아 사카랄리스), 명충 나방 유충(LSCB, 엘라스모팔푸스 리그노셀루스), 왕담배 밤나방(CEW, 헬리코베르파 제아), 회색 담배 나방(TBW, 헬리오티스 비레센스), 대두 애벌레(SBL, 크리소데익시스 인클루덴스), 블랙 멸강나방(BLAW, 스포돕테라 코스미오이데스), 남방 멸강나방(SAW, 스포돕테라 에리다니아), 가을멸강충(FAW, 스포돕테라 프루기페르다), 파밤나방(BAW, 스포돕테라 엑시구아), 올드 월드 볼웜(OBW, 헬리코베르파 아르미게라), 오리엔탈 리프웜(OLW, 스포돕테라 리투라), 분홍 솜벌레(PBW, 펙티노포라 고시피엘라), 검거세미 나방(BCW, 아그로티스 입실론), 남서부 조명충 나방(SWCB, 디아트라에아 그란디오셀라), 스팟티드 볼웜(SBW, 에아리아스 비텔라), 및 유럽 조명충 나방(ECB, 오스트리니아 누빌랄리스)에 대한 활성에 대해서 검정하였다. 왕담배 밤나방(CEW, 헬리코베르파 제아)은 대두 팟웜(SPW) 및 목화씨 벌레(CBW)라고도 지칭된다. 사멸률 및 발육 저지 점수뿐만 아니라 MIC50 점수의 조합을 통해서 활성을 결정하였다. MIC50은 죽은 유충 및 L1 유충(제2 영기로의 탈피에 실패한 유충) 모두가 점수에서 고려되는 탈피 억제 농도를 말한다. 표 2는 각각의 키메릭 살곤충 단백질의 활성을 보여준다. '+' 표시는 특정 곤충 해충에 대해서 관찰된 활성을 나타낸다.
상기 표 2로부터 알 수 있는 바와 같이, 키메릭 살곤충 단백질의 대부분이 1종 이상의 인시류 해충 종에 대해서 활성을 나타내었다.
실시예 3
식물에서의 발현을 위한 키메릭 살곤충 단백질을 암호화하는 유전자의 합성
본 실시예는 식물에서의 발현을 위한 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드의 합성을 보여준다.
식물에서의 키메릭 살곤충 단백질의 발현에서 사용하기 위해서 합성 암호화 서열을 구성하였다. 미국 특허 제5,500,365호에 일반적으로 기술된 방법에 따라서 합성 서열을 설계하고, 합성하여, 키메릭 살곤충 단백질의 아미노산 서열을 보존하면서, 특정 해로운 문제 서열, 예컨대 ATTTA 및 A/T 풍부 식물 폴리아데닐화 서열을 회피하였다. 식물에서의 발현을 위한 키메릭 살곤충 단백질을 암호화하는 이러한 유전자에 대한 뉴클레오타이드 서열을 표 3에 열거한다.
실시예 4
식물에서의 키메릭 살곤충 단백질의 발현을 위한 발현 카세트
본 실시예는 키메릭 살곤충 단백질을 암호화하는 식물에서 사용하기 위해서 설계된 폴리뉴클레오타이드 서열을 포함하는 발현 카세트의 구성을 보여준다.
표 3에 제공된 식물 발현을 위해서 설계된 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드 서열을 사용하여 다양한 식물 발현 카세트를 구성하였다. 이와 같은 발현 카세트는 식물 원형질에서의 일시적인 발현 또는 식물 세포의 형질변환에 유용하다. 전형적인 발현 카세트는 세포 내 단백질의 궁극적인 배치에 관련하여 고안되었다. 발현 카세트 중 하나의 세트는 단백질이 번역되어 세포질 내에 남아있을 수 있도록 하는 방식으로 고안되었다. 발현 카세트의 다른 세트는 엽록체 또는 색소체와 같은 세포의 세포 기관의 표적화를 가능하게 하기 위하여 독소 단백질과 인접하는 수송 펩타이드를 갖도록 설계되었다. 모든 발현 카세트는, 다수의 프로모터 요소, 인핸서 요소, 또는 당업자에게 공지된 트랜스진(transgene)의 발현을 북돋우도록 작동가능하게 연결된 다른 발현 요소로 구성될 수 있는, 프로모터를 가지는 5' 말단에서 시작하도록 설계되었다. 프로모터 서열은 프로모터에 대하여 보통 하나 이상의 리더 서열 3'과 연속적으로 이어졌다. 인트론 서열은 보통 리더 서열에 대하여 3'에 제공되어 트랜스진의 발현을 개선시켰다. 독소에 대하여 암호화 서열 또는 독소에 대하여 수송 펩타이드 및 암호화 서열은 보통 작동가능하게 연결된 프로모터, 리더 및 인트론 배역에 대하여 3'에 위치하였다. 3'UTR 서열은 보통 암호화 서열의 3'에서 제공되어 전사의 종결을 용이하게 하고 생성된 전사체의 폴리아데닐화에 중요한 서열을 제공하였다. 상기 기재된 요소 모두, 종종 발현 카세트의 구조에 제공된 추가적인 서열에 작동가능하게 연결되고, 순차적으로 배열되었다.
실시예 5
안정하게 형질전환된 옥수수에서의 키메릭 살곤충 단백질의 인시류 활성
본 실시예는 옥수수 식물에서 발현되어, 각각의 옥수수 곤충 해충에 대한 먹이로서 제공되는 경우 인시류 해충에 대해서 키메릭 살곤충 단백질이 나타내는 저해 활성을 보여준다.
옥수수 품종 LH244를 아그로박테리움-매개된 형질전환 방법을 사용하여 실시예 4에 기술된 이원(binary) 형질전환 벡터를 사용하여 형질전환시켰다. 당업계에 공지된 방법에 의해 형질전환된 세포를 유도하여 식물을 형성하였다. 미국 특허 제8,344,207호에 기술된 것과 유사하게 식물 잎 디스크를 사용하는 생물학적 검정을 수행하였다. 형질전환되지 않은 LH244 식물을 사용하여 음성 대조군으로서 사용될 조직을 수득하였다. 각각의 이원 벡터로부터의 다중 트랜스제닉 이벤트를 왕담배 밤나방(CEW, 헬리코베르파 제아), 가을멸강충(FAW, 스포돕테라 프루기페르다), 검거세미 나방(BCW, 아그로티스 입실론) 및 남서부 조명충 나방(SWCB, 디아트라에아 그란디오셀라)에 대해서 평가하였다.
R0 세대 유전자 이식 식물 및 F1 세대 유전자 이식 식물에 대해서 잎 디스크 생물학적 검정을 수행하였다. 또한, 인시류 곤충 해충이 침입한 특정 키메릭 살곤충 단백질을 발현시킨 전체 트랜스제닉 F1 식물에 대해서 잎 손상 순위를 평가하였다. TIC860 및 TIC868을 발현하는 F1 트랜스제닉 이벤트를 또한 FAW, CEW, 및 SWCB에 대해서 필드(field)에서의 활성에 대해서 평가하였다. 검정 결과를 표 4에 나타낸다. '+' 표시는 특정 곤충 해충에 대해서 관찰된 활성을 나타낸다. 표 4에서 알 수 있는 바와 같이, 키메릭 살곤충 단백질의 대부분 및 키메릭 살곤충 단백질 변종의 다수는 1종 이상의 인시류 해충 종에 대해서 활성을 나타내었다.
실시예 6
안정하게 형질전환된 대두에서의 키메릭 살곤충 단백질의 인시류 활성
본 실시예는 대두 식물에서 발현되어, 각각의 곤충 해충에 대한 먹이로서 제공되는 경우 인시류 해충에 대해서 키메릭 살곤충 단백질이 나타내는 저해 활성을 보여준다.
선별된 키메릭 살곤충 단백질에 대한 암호화 서열을 식물 발현을 위해서 재설계하고, 이원 식물 형질전환 벡터에 클로닝하고, 그것을 사용하여 대두 식물 세포를 형질전환시켰다. 식물 형질전환 벡터는 실시예 4에 기술된 바와 같은 키메릭 살곤충 단백질의 발현을 위한 제1 트랜스진 카세트 및 스펙티노마이신 선별을 사용한 트랜스제닉 식물 세포의 선별을 위한 제2 트랜스진 카세트를 포함하였다. 일부 예에서, 예컨대 TIC1100, TIC860 및 TIC836의 경우에, 엽록체 트랜지트 펩타이드 암호화 서열을 키메릭 살곤충 암호화 서열에 작동 가능하게 연결시켰다. 색소체 표적화된 그리고 표적화되지 않은 TIC1100, TIC860 및 TIC836을 사용하여 검정을 수행하였다. 하기 표 5는 안정하게 형질전환된 대두에서의 발현을 위해서 사용된 키메릭 살곤충 단백질 및 TIC867 변종 키메릭 살곤충 단백질 및 관련 암호화 서열을 보여준다.
아그로박테리움-매개된 형질전환에 의해서 상기에 기술된 이원 형질전환 벡터를 사용하여 대두 식물 세포를 형질전환시켰다. 생성된 형질전환된 식물 세포를 전체 대두 식물을 형성하도록 유도하였다. 잎 조직을 수확하고, 실시예 5에 기술된 바와 같은 생물학적 검정에서 사용하거나, 또는 대안적으로, 동결 건조된 조직을 생물학적 검정을 위한 곤충 먹이로 사용하였다. FAW, 남방 멸강나방(SAW, 스포돕테라 에리다니아), 대두 유충(SBL, 크리소데익시스 인클루덴스), 대두 팟웜(SPW, 헬리코베르파 제아), 벨벳콩 자나방(VBC, 안티카르시아 겜마탈리스), 회색 담배 나방(TBW, 헬리오티스 비레센스), 블랙 멸강나방(BLAW, 스포돕테라 코스미오이데스), 명충 나방 애벌레(LSCB, 엘라스모팔푸스 리그노셀루스) 및 올드 월드 볼웜(OBW, 헬리코베르파 아르미게라)에 대해서 생물학적 검정을 수행하였다.
표 5는 R0 세대 식물에서의 각각의 살곤충 단백질에 대한 인시류의 선택된 종에 대한 활성을 보여주는데, 여기서 '+'는 활성을 나타낸다. 표 5에서 알 수 있는 바와 같이, 안정하게 형질전환된 대두에서 발현된 키메릭 살곤충 단백질 각각은 다수의 인시류 종에 대해서 활성을 나타내었다. 특히, TIC867 변종인 TIC867_23이 SPW에 대해서 활성을 나타낸 것이 주목된다.
선택된 형질전환된 이벤트를 자가 수분하게 하고, 생성된 종자를 성장시켰다. R1 세대 식물로부터 잎 조직을 수확하여, 섭취 생물학적 검정에서 사용하였다. TIC1100, TIC860, TIC867, TIC868, TIC869 및 TIC836을 발현하는 R1 식물을 SAW, SBL, SPW 및 VBC에 대한 활성에 대해서 검정하였다. 표 6은 이 시험에서 관찰된 활성을 나타낸다. '+' 표시는 특정 곤충 해충에 대해서 관찰된 활성을 나타낸다. 표 6에 나타낸 바와 같이, R1 세대 식물로부터의 발현된 키메릭 살곤충 단백질 대부분은 1종 이상의 인시류 종에 대해서 활성을 나타내었다.
표 7은 TIC1100, TIC860, 및 TIC836을 발현하는 안정하게 형질전환된 R1 세대 대두 식물을 사용하여 스크린 하우스에서 수행된 필드 시험의 결과를 나타낸다. 스크린 하우스에서 식물 침입에 사용된 SAW, SBL 및 SPW를 포함한다. 대두 식물에서 탈엽이 15% 이하일 때 저항성이라고 정의하였다. 이러한 케이지 시험에서 관찰된 저항성은 표 6에 나타낸 R1 세대 대두 잎 조직 검정에서 관찰된 저항성과 일치한다. '+' 표시는 특정 곤충 해충에 대해서 관찰된 활성을 나타낸다.
TIC867 및 TIC869를 발현하는 안정하게 형질전환된 R1 세대 대두 식물을 사용한 스크린 하우스에서의 필드 시험을 또한 아르헨티나의 상이한 두 지역, 즉 아세베도(Acevedo) 및 폰테주엘라(Fontezuela)에서 수행하였다. 스크린 하우스에서 식물 침입에 사용된 종은 남미 목화씨 벌레(South American bollworm)(SABW, 헬리코베르파 겔로토페온), VBC, BLAW, 및 해바라기 애벌레(SFL, 라치플루시아 누)를 포함한다. 대두 식물에서 탈엽이 15% 이하일 때 저항성이라고 정의하였다. 하기 표 8은 관찰된 저항성을 보여준다. '+' 표시는 특정 곤충 해충에 대해서 관찰된 활성을 나타낸다. 표 8에 나타낸 바와 같이, TIC867을 발현하는 트랜스제닉 대두 식물은 BLAW 및 VBC에 저항성을 나타내었다. TIC869를 발현하는 트랜스제닉 대두 식물은 SABW, SFL, BLAW 및 VBC에 저항성을 나타내었다.
실시예 7
안정하게 형질전환된 목화에서의 키메릭 살곤충 단백질의 인시류 활성
본 실시예는 목화 식물에서 발현되어, 각각의 곤충 해충에 대한 먹이로서 제공되는 경우 인시류 해충에 대한 키메릭 살곤충 단백질이 나타내는 저해 활성을 보여준다.
선택된 키메릭 살곤충 단백질에 대한 암호화 서열을 식물 발현을 위해서 재설계하고, 이원 식물 형질전환 벡터에 클로닝하고, 그것을 사용하여 목화 식물 세포를 형질전환시켰다. 생성된 이원 벡터는 실시예 4에 기술된 것과 유사하였고, 그것을 사용하여 색소체 표적화되거나, 색소체 표적화되지 않은 TIC860(암호화 서열: 서열번호 6; 단백질 서열: 서열번호 7), TIC867(암호화 서열: 서열번호 9; 단백질 서열: 서열번호 10), TIC868(암호화 서열: 서열번호 27; 단백질 서열: 서열번호 28) 및 TIC867_23(암호화 서열: 서열번호 20; 단백질 서열: 서열번호 23)을 발현시켰다.
아그로박테리움-매개된 형질전환 방법에 의해서 목화 식물 세포를 형질전환시켰다. 형질전환된 목화 세포를 전체 식물을 형성하도록 유도하였다. 목화 잎 조직을 실시예 5에 기술된 바와 같이 목화씨 벌레(CBW, 헬리코베르파 제아), FAW, TBW 및 SBL에 대한 생물학적 검정에서 사용하였다. 표 9는 안정하게 형질전환된 R0 세대 목화에서 TIC860, TIC867, 및 TIC868에 대한 이러한 인시류 종에 대해서 관찰된 활성을 보여주며, 여기서 '+'는 활성을 나타낸다. 표 9에서 알 수 있는 바와 같이, TIC860, TIC867, 및 TIC868은 안정하게 형질전환된 R0 세대 목화에서 2종 이상의 인시류 해충 종에 대해서 활성을 나타내었다.
선택된 트랜스제닉 이벤트를 사용하여 R1 종자를 생산하였다. TIC860, TIC867, 및 TIC868을 발현하는 R1 식물을 CBW, FAW, TBW, 및 SBL에 대한 저항성에 대해서 검정하였다. 잎 조직, 봉오리 조직 및 꼬투리 조직을 검정에서 사용하였다. 표 10은 이 시험에서 관찰된 활성을 나타낸다. '+' 표시는 특정 해충에 대해서 관찰된 활성을 나타낸다. 표 10에서 입증되는 바와 같이, TIC860은 잎 조직에서 FAW에 대해서 활성을 나타내었다. 추가로, 키메릭 살곤충 단백질 TIC867은 잎 조직, 봉오리 조직 및 꼬투리 조직에서 CBW 및 FAW에 대해서 활성을 나타내었을 뿐만 아니라, 잎에서는 TBW 및 SBL에 대해서 활성을 나타내었다. 키메릭 살곤충 단백질 TIC868은 잎 조직, 봉오리 조직 및 꼬투리 조직에서 FAW에 대해서 활성을 나타내었을 뿐만 아니라, 잎에서는 TBW 및 SBL에 대해서 활성을 나타내었다.
본원에 개시되고 청구된 조성물 모두는 본 개시물을 참고하여 과도한 실험 없이 제조되고 성취될 수 있다. 본 발명의 조성물이 상기 예시적인 실시형태와 관련하여 기술되어 있지만, 본 발명의 충실한 개념, 사상 및 범주를 벗어나지 않으면서 변형, 변화, 개질 및 변경이 본원에 기술된 조성물에 적용될 수 있음은 당업자에게 자명할 것이다. 보다 구체적으로, 화학적 및 생리학적으로 모두 관련된 특정 제제가 본 명세서에 기재된 제제를 대체할 수 있는 한편, 동일 또는 유사한 결과가 얻어질 것이다. 당업자에게 명백한 이와 같은 유사한 대체 및 변형은 모두 첨부된 청구범위에 의해 한정되는 바와 같이 본 발명의 사상, 범주, 및 개념 내에 있는 것으로 여겨진다.
본 명세서에 인용된 모든 공개물 및 공개된 특허 문헌은 각각의 개별 공개물 또는 특허 출원이 구체적이고 개별적으로 참고로 포함된 것처럼 동일한 정도로 참고로 본원에 포함된다.
SEQUENCE LISTING
<110> Monsanto Technology LLC
Baum, James A
Cerruti, Thomas A
Dart, Crystal L
English, Leigh H
Fu, Xiaoran
Guzov, Victor M
Howe, Arlene R
Morgenstern, Jay P
Roberts, James K
Salvador, Sara A
Wang, Jinling
<120> Novel Chimeric Insecticidal Proteins Toxic or Inhibitory to
Lepidopteran Pests
<130> P34230WO00/0022270.00098
<150> US 62/064989
<151> 2014-10-16
<160> 53
<170> PatentIn version 3.5
<210> 1
<211> 3570
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC1100.
<400> 1
atggagatag tgaataatca gaatcaatgc gtgccttata attgtttgaa taatcccgaa 60
atcgaaatat tagaaggcgg aagaatatca gttggtaata ccccaattga tatttctctt 120
tcgcttactc agtttctttt gagtgaattt gtcccaggtg cggggtttgt attaggatta 180
attgatttaa tatggggatt tgtaggtcct tcccaatggg acgcatttct tgctcaagtg 240
gaacagttaa ttaaccaaag aatagcagaa gctgtaagaa atacagcaat tcaggaatta 300
gagggaatgg cacgggttta tagaacctat gctactgctt ttgctgagtg ggaaaaagct 360
cctgatgacc cagagctaag agaagcacta cgtacacaat ttacagcaac tgagacttat 420
ataagtggaa gaatatccgt tttaaaaatt caaacttttg aagtacagct gttatcagtg 480
tttgcccaag ctgcaaattt acatttatct ttattaagag acgttgtgtt ttttgggcaa 540
agatggggtt tttcaacgac aaccgtaaat aattactaca atgatttaac agaagggatt 600
agtacctata cagattatgc tgtacgctgg tacaatacgg gattagaacg tgtatgggga 660
ccggattcta gagattgggt aaggtataat caatttagaa gagaattaac actaactgta 720
ttagatatcg ttgctctgtt cccgaattat gatagtagaa gatatccaat tcgaacagtt 780
tcccaattaa caagagaaat ttatacaaac ccagtattag aaaattttga tggtagtttt 840
cgaggctcgg ctcagggcat agaaagaagt attaggagtc cacatttgat ggatatactt 900
aacagtataa ccatctatac ggatgctcat aggggttatt attattggtc agggcatcaa 960
ataatggctt ctcctgtcgg tttttcgggg ccagaattca cgtttccgct atatggaacc 1020
atgggaaatg cagctccaca acaacgtatt gttgctcaac taggtcaggg cgtgtataga 1080
acattatcgt ccactttata tagaagacct tttaatatag ggataaataa tcaacaacta 1140
tctgttcttg acgggacaga atttgcttat ggaacctcct caaatttgcc atccgctgta 1200
tacagaaaaa gcggaacggt agattcgctg gatgaaatac cgccacagaa taacaacgtg 1260
ccacctaggc aaggatttag tcatcgatta agccatgttt caatgtttcg ttcaggcttt 1320
agtaatagta gtgtaagtat aataagagct cctatgttct cttggataca tcgtagtgct 1380
gaatttaata atataattgc atcggatagt attaatcaaa tacctttagt gaaaggattt 1440
agagtttggg ggggcacctc tgtcattaca ggaccaggat ttacaggagg ggatatcctt 1500
cgaagaaata cctttggtga ttttgtatct ctacaagtca atattaattc accaattacc 1560
caaagatacc gtttaagatt tcgttacgct tccagtaggg atgcacgagt tatagtatta 1620
acaggagcgg catccacagg agtgggaggc caagttagtg taaatatgcc tcttcagaaa 1680
actatggaaa taggggagaa cttaacatct agaacattta gatataccga ttttagtaat 1740
cctttttcat ttagagctaa tccagatata attgggataa gtgaacaacc tctatttggt 1800
gcaggttcta ttagtagcgg tgaactttat atagataaaa ttgaaattat tctagcagat 1860
gcaacatttg aagcagaatc tgatttagaa agagcgcaga aggcggtgaa tgcgctgttt 1920
acgtctacaa accaactagg gctaaaaaca aatgtaacgg attatcatat tgatcaagtg 1980
tccaatttag ttacgtattt atcggatgaa ttttgtctgg atgaaaagcg agaattgtcc 2040
gagaaagtca aacatgcgaa gcgactcagt gatgaacgca atttactcca agattcaaat 2100
ttcaaagaca ttaataggca accagaacgt gggtggggcg gaagtacagg gattaccatc 2160
caaggagggg atgacgtatt taaagaaaat tacgtcacac tatcaggtac ctttgatgag 2220
tgctatccaa catatttgta tcaaaaaatc gatgaatcaa aattaaaagc ctttacccgt 2280
tatcaattaa gagggtatat cgaagatagt caagacttag aaatctattt aattcgctac 2340
aatgcaaaac atgaaacagt aaatgtgcca ggtacgggtt ccttatggcc gctttcagcc 2400
caaagtccaa tcggaaagtg tggagagccg aatcgatgcg cgccacacct tgaatggaat 2460
cctgacttag attgttcgtg tagggatgga gaaaagtgtg cccatcattc gcatcatttc 2520
tccttagaca ttgatgtagg atgtacagac ttaaatgagg acctaggtgt atgggtgatc 2580
tttaagatta agacgcaaga tgggcacgca agactaggga atctagagtt tctcgaagag 2640
aaaccattag taggagaagc gctagctcgt gtgaaaagag cggagaaaaa atggagagac 2700
aaacgtgaaa aattggaatg ggaaacaaat atcgtttata aagaggcaaa agaatctgta 2760
gatgctttat ttgtaaactc tcaatatgat caattacaag cggatacgaa tattgccatg 2820
attcatgcgg cagataaacg tgttcatagc attcgagaag cttatctgcc tgagctgtct 2880
gtgattccgg gtgtcaatgc ggctattttt gaagaattag aagggcgtat tttcactgca 2940
ttctccctat atgatgcgag aaatgtcatt aaaaatggtg attttaataa tggcttatcc 3000
tgctggaacg tgaaagggca tgtagatgta gaagaacaaa acaaccaacg ttcggtcctt 3060
gttgttccgg aatgggaagc agaagtgtca caagaagttc gtgtctgtcc gggtcgtggc 3120
tatatccttc gtgtcacagc gtacaaggag ggatatggag aaggttgcgt aaccattcat 3180
gagatcgaga acaatacaga cgaactgaag tttagcaact gcgtagaaga ggaaatctat 3240
ccaaataaca cggtaacgtg taatgattat actgtaaatc aagaagaata cggaggtgcg 3300
tacacttctc gtaatcgagg atataacgaa gctccttccg taccagctga ttatgcgtca 3360
gtctatgaag aaaaatcgta tacagatgga cgaagagaga atccttgtga atttaacaga 3420
gggtataggg attacacgcc actaccagtt ggttatgtga caaaagaatt agaatacttc 3480
ccagaaaccg ataaggtatg gattgagatt ggagaaacgg aaggaacatt tatcgtggac 3540
agcgtggaat tactccttat ggaggaatga 3570
<210> 2
<211> 3570
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC1100.
<400> 2
atggagattg tgaacaacca gaaccagtgc gttccttaca actgcttgaa caaccctgag 60
attgagattc ttgagggtgg tagaatttct gttggcaaca ctcctattga catctctttg 120
agtttgactc aattcttgtt gagtgagttc gttcctggtg ctggtttcgt cttgggtttg 180
attgatttga tttggggttt cgttggtcct agtcaatggg atgctttctt ggctcaagtt 240
gagcaattga ttaaccagag gatcgctgag gctgtgagga acactgctat tcaagagttg 300
gagggtatgg ctagagttta cagaacttac gctactgctt tcgctgagtg ggagaaggct 360
cctgatgacc ctgagttgag ggaggctttg agaactcaat tcactgctac tgagacttac 420
atcagtggta gaatcagtgt cttgaagatt caaactttcg aggttcaatt gctttctgtg 480
ttcgctcaag ctgcaaactt gcacttgtct ttgcttagag atgttgtgtt ctttggtcaa 540
agatggggtt tctccactac taccgtgaac aattactaca acgatttgac tgagggtatt 600
tctacttaca ctgattacgc tgttagatgg tacaacactg gtttggagag agtttggggt 660
ccagattcca gagattgggt cagatacaac cagttcagaa gggagttgac tttgactgtc 720
ttggacattg ttgctctctt ccctaactac gatagtcgtc gttaccctat tagaactgtt 780
tctcaactta ctagggaaat ctacactaac cctgttcttg agaacttcga tggtagtttc 840
cgtggtagtg ctcaagggat tgagcgttct attcgttctc ctcatcttat ggacattctt 900
aactctatta ctatctacac tgatgctcat cgtggttact attactggtc tggtcatcaa 960
attatggcta gtcctgttgg tttcagtggt cctgagttca ctttccctct ttacggtact 1020
atgggcaacg ctgcacctca acagaggatc gttgctcaac ttggtcaagg tgtttacagg 1080
actctttctt caacccttta caggcgtcct ttcaacattg ggatcaacaa ccagcagctt 1140
tctgttcttg atggaaccga gttcgcttac ggaacctctt caaaccttcc tagtgctgtt 1200
tacaggaagt ctggaaccgt tgacagtctt gatgagattc caccgcagaa caataacgtt 1260
ccacccaggc aaggcttcag tcataggctt tctcatgttt ctatgttccg ctctggattc 1320
agcaactctt cagtttctat tatcagggct ccaatgttct cgtggattca taggtctgcc 1380
gagttcaaca acattatcgc ttccgatagc attaaccaga ttccacttgt taagggattc 1440
cgtgtttggg gaggcacctc tgttattacc ggaccaggct tcaccggagg cgacattctt 1500
cgtcgtaaca ccttcggaga tttcgtttca cttcaagtga acattaactc accaatcacc 1560
cagcgctaca ggcttcgctt ccgctacgca tcatccaggg atgcaagggt gatcgtgctt 1620
accggagcag cctcaaccgg agtgggaggc caagtgagcg tgaacatgcc acttcagaag 1680
acgatggaga tcggcgagaa ccttacctca agaacctttc gttacaccga tttcagcaac 1740
ccattcagct ttcgtgcaaa cccagacatc atagggatct cagagcagcc actgtttgga 1800
gctggatcaa tctcatccgg agagctttac atcgacaaga tcgagatcat actcgcagat 1860
gcaaccttcg aggctgagag cgatctggag cgtgcacaga aggcagtgaa cgcactcttt 1920
acctctacca accagctcgg actcaagacc aacgtgaccg attaccacat cgaccaagtg 1980
agcaacctcg tgacctacct ctcagatgag ttctgcttgg atgagaaacg cgaactcagc 2040
gagaaggtga agcacgcaaa gcgtctctca gatgagcgta acctcctcca ggatagcaat 2100
ttcaaggaca tcaatcgtca gccagagcgt ggatggggag gctcaaccgg aatcaccatc 2160
cagggaggcg atgatgtgtt taaggagaat tacgtgacac tctccggaac attcgatgag 2220
tgctacccaa catacctcta tcagaagatc gacgagtcca agctcaaggc gttcacccgt 2280
tatcagctcc gtggctacat cgaggatagt caagacctgg aaatctacct catccgctac 2340
aatgcaaagc acgagacagt gaatgtgcca ggaacaggct ccctctggcc actctccgca 2400
cagtctccaa tcggcaagtg cggcgagcca aatcgctgcg cgccacacct ggagtggaat 2460
cccgacctgg actgctcctg ccgcgacggc gagaagtgcg cccaccactc ccaccacttt 2520
agcctggaca tcgacgtggg ctgtacagac ctgaatgagg atctgggcgt gtgggtgatc 2580
tttaagatca agacacagga cggccacgcc cgcctgggca atctggagtt tctggaggag 2640
aagcctctgg tgggcgaagc cctggcccgc gtgaagcgcg ccgagaagaa atggcgcgac 2700
aaacgcgaga aactggaatg ggaaacaaac atcgtgtaca aagaagccaa agaatccgtg 2760
gacgccctat ttgtgaactc ccagtatgac cagctacagg ccgacacaaa catcgcgatg 2820
atccacgctg cggacaagcg cgtgcactcc atacgcgaag cctatctacc cgaactatcc 2880
gtgatacccg gcgtcaatgc cgcgatcttt gaagaattgg aaggccgcat cttcacagcc 2940
tttagcctct atgacgcccg aaatgtcatc aagaatggcg actttaacaa tgggctatcc 3000
tgttggaatg tcaaagggca cgtggacgtc gaagagcaga acaatcagcg atccgtctta 3060
gtcgtacccg aatgggaagc cgaagtctcc caggaagtcc gagtctgtcc tggtagaggt 3120
tacatcttga gagtgactgc ttacaaggag ggttacggtg agggatgcgt gactattcac 3180
gagattgaga acaacactga tgagttgaag ttcagtaact gcgtggagga ggaaatctac 3240
cccaacaaca ctgtgacttg taacgattac accgtgaacc aggaggaata cggaggcgct 3300
tacacctcca gaaaccgtgg atacaatgag gctccctcgg tccccgctga ttatgcctcc 3360
gtctatgagg agaagtccta caccgatgga aggcgcgaga atccctgcga gttcaatcgc 3420
ggctatcgag actacactcc gctacccgtt ggctatgtca caaaggaact ggaatacttc 3480
ccggaaacag acaaagtctg gatcgaaatc ggcgaaacag aagggacgtt catagtcgat 3540
agcgtagaac ttctccttat ggaagaatga 3570
<210> 3
<211> 3570
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC1100.
<400> 3
atggagattg tgaacaacca gaaccagtgc gttccttaca actgcttgaa caaccctgag 60
attgagattc ttgagggtgg tagaatttct gttggcaaca ctcctattga catctctttg 120
agtttgactc aattcttgtt gagtgagttc gttcctggtg ctggtttcgt cttgggtttg 180
attgatttga tttggggttt cgttggtcct agtcaatggg atgctttctt ggctcaagtt 240
gagcaattga ttaaccagag gatcgctgag gctgtgagga acactgctat tcaagagttg 300
gagggtatgg ctagagttta cagaacttac gctactgctt tcgctgagtg ggagaaggct 360
cctgatgacc ctgagttgag ggaggctttg agaactcaat tcactgctac tgagacttac 420
atcagtggta gaatcagtgt cttgaagatt caaactttcg aggttcaatt gctttctgtg 480
ttcgctcaag ctgcaaactt gcacttgtct ttgcttagag atgttgtgtt ctttggtcaa 540
agatggggtt tctccactac taccgtgaac aattactaca acgatttgac tgagggtatt 600
tctacttaca ctgattacgc tgttagatgg tacaacactg gtttggagag agtttggggt 660
ccagattcca gagattgggt cagatacaac cagttcagaa gggagttgac tttgactgtc 720
ttggacattg ttgctctctt ccctaactac gatagtcgtc gttaccctat tagaactgtt 780
tctcaactta ctagggaaat ctacactaac cctgttcttg agaacttcga tggtagtttc 840
cgtggtagtg ctcaagggat tgagcgttct attcgttctc ctcatcttat ggacattctt 900
aactctatta ctatctacac tgatgctcat cgtggttact attactggtc tggtcatcaa 960
attatggcta gtcctgttgg tttcagtggt cctgagttca ctttccctct ttacggtact 1020
atgggcaacg ctgcacctca acagaggatc gttgctcaac ttggtcaagg tgtttacagg 1080
actctttctt caacccttta caggcgtcct ttcaacattg ggatcaacaa ccagcagctt 1140
tctgttcttg atggaaccga gttcgcttac ggaacctctt caaaccttcc tagtgctgtt 1200
tacaggaagt ctggaaccgt tgacagtctt gatgagattc caccgcagaa caataacgtt 1260
ccacccaggc aaggcttcag tcataggctt tctcatgttt ctatgttccg ctctggattc 1320
agcaactctt cagtttctat tatcagggct ccaatgttct cgtggattca taggtctgcc 1380
gagttcaaca acattatcgc ttccgatagc attaaccaga ttccacttgt taagggattc 1440
cgtgtttggg gaggcacctc tgttattacc ggaccaggct tcaccggagg cgacattctt 1500
cgtcgtaaca ccttcggaga tttcgtttca cttcaagtga acattaactc accaatcacc 1560
cagcgctaca ggcttcgctt ccgctacgca tcatccaggg atgcaagggt gatcgtgctt 1620
accggagcag cctcaaccgg agtgggaggc caagtgagcg tgaacatgcc acttcagaag 1680
acgatggaga tcggcgagaa ccttacctca agaacctttc gttacaccga tttcagcaac 1740
ccattcagct ttcgtgcaaa cccagacatc atagggatct cagagcagcc actgtttgga 1800
gctggatcaa tctcatccgg agagctttac atcgacaaga tcgagatcat actcgcagat 1860
gcaaccttcg aggctgagag cgatctggag cgtgcacaga aggcagtgaa cgcactcttt 1920
acctctacca accagctcgg actcaagacc aacgtgaccg attaccacat cgaccaagtg 1980
agcaacctcg tgacctacct ctcagatgag ttctgcttgg atgagaaacg cgaactcagc 2040
gagaaggtga agcacgcaaa gcgtctctca gatgagcgta acctcctcca ggatagcaat 2100
ttcaaggaca tcaatcgtca gccagagcgt ggatggggag gctcaaccgg aatcaccatc 2160
cagggaggcg atgatgtgtt taaggagaat tacgtgacac tctccggaac attcgatgag 2220
tgctacccaa catacctcta tcagaagatc gacgagtcca agctcaaggc gttcacccgt 2280
tatcagctcc gtggctacat cgaggatagt caagacctgg aaatctacct catccgctac 2340
aatgcaaagc acgagacagt gaatgtacca ggaacaggct ccctctggcc actctccgca 2400
cagtctccaa tcggcaagtg cggcgagcca aatcgctgcg cgccacacct ggagtggaat 2460
cccgacctgg actgctcctg ccgcgacggc gagaagtgcg cccaccactc ccaccacttt 2520
agcctggaca tcgacgtggg ctgtacagac ctgaatgagg atctgggcgt gtgggtgatc 2580
tttaagatca agacacagga cggccacgcc cgcctgggca atctggagtt tctggaggag 2640
aagcctctgg tgggcgaagc cctggcccgc gtgaagcgcg ccgagaagaa atggcgcgac 2700
aaacgcgaga aactggaatg ggaaacaaac atcgtgtaca aagaagccaa agaatccgtg 2760
gacgccctat ttgtgaactc ccagtatgac cagctacagg ccgacacaaa catcgcgatg 2820
atccacgctg cggacaagcg cgtgcactcc atacgcgaag cctatctacc cgaactatcc 2880
gtgatacccg gcgtcaatgc cgcgatcttt gaagaattgg aaggccgcat cttcacagcc 2940
tttagcctct atgacgcccg aaatgtcatc aagaatggcg actttaacaa tgggctatcc 3000
tgttggaatg tcaaagggca cgtggacgtc gaagagcaga acaatcagcg atccgtctta 3060
gtcgtacccg aatgggaagc cgaagtctcc caggaagtcc gagtctgtcc tggtagaggt 3120
tacatcttga gagtgactgc ttacaaggag ggttacggtg agggatgcgt gactattcac 3180
gagattgaga acaacactga tgagttgaag ttcagtaact gcgtggagga ggaaatctac 3240
cccaacaaca ctgtgacttg taacgattac accgtgaacc aggaggaata cggaggcgct 3300
tacacctcca gaaaccgtgg atacaatgag gctccctcgg tccccgctga ttatgcctcc 3360
gtctatgagg agaagtccta caccgatgga aggcgcgaga atccctgcga gttcaatcgc 3420
ggctatcgag actacactcc gctacccgtt ggctatgtca caaaggaact ggaatacttc 3480
ccggaaacag acaaagtctg gatcgaaatc ggcgaaacag aagggacgtt catagtcgat 3540
agcgtagaac ttctccttat ggaagaatga 3570
<210> 4
<211> 1189
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein TIC1100.
<400> 4
Met Glu Ile Val Asn Asn Gln Asn Gln Cys Val Pro Tyr Asn Cys Leu
1 5 10 15
Asn Asn Pro Glu Ile Glu Ile Leu Glu Gly Gly Arg Ile Ser Val Gly
20 25 30
Asn Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser
35 40 45
Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Ile Asp Leu Ile
50 55 60
Trp Gly Phe Val Gly Pro Ser Gln Trp Asp Ala Phe Leu Ala Gln Val
65 70 75 80
Glu Gln Leu Ile Asn Gln Arg Ile Ala Glu Ala Val Arg Asn Thr Ala
85 90 95
Ile Gln Glu Leu Glu Gly Met Ala Arg Val Tyr Arg Thr Tyr Ala Thr
100 105 110
Ala Phe Ala Glu Trp Glu Lys Ala Pro Asp Asp Pro Glu Leu Arg Glu
115 120 125
Ala Leu Arg Thr Gln Phe Thr Ala Thr Glu Thr Tyr Ile Ser Gly Arg
130 135 140
Ile Ser Val Leu Lys Ile Gln Thr Phe Glu Val Gln Leu Leu Ser Val
145 150 155 160
Phe Ala Gln Ala Ala Asn Leu His Leu Ser Leu Leu Arg Asp Val Val
165 170 175
Phe Phe Gly Gln Arg Trp Gly Phe Ser Thr Thr Thr Val Asn Asn Tyr
180 185 190
Tyr Asn Asp Leu Thr Glu Gly Ile Ser Thr Tyr Thr Asp Tyr Ala Val
195 200 205
Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg
210 215 220
Asp Trp Val Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val
225 230 235 240
Leu Asp Ile Val Ala Leu Phe Pro Asn Tyr Asp Ser Arg Arg Tyr Pro
245 250 255
Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val
260 265 270
Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu
275 280 285
Arg Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr
290 295 300
Ile Tyr Thr Asp Ala His Arg Gly Tyr Tyr Tyr Trp Ser Gly His Gln
305 310 315 320
Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro
325 330 335
Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala
340 345 350
Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg
355 360 365
Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp
370 375 380
Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val
385 390 395 400
Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln
405 410 415
Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His
420 425 430
Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile
435 440 445
Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Glu Phe Asn Asn
450 455 460
Ile Ile Ala Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
465 470 475 480
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
485 490 495
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
500 505 510
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
515 520 525
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
530 535 540
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
545 550 555 560
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
565 570 575
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
580 585 590
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
595 600 605
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu
610 615 620
Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe
625 630 635 640
Thr Ser Thr Asn Gln Leu Gly Leu Lys Thr Asn Val Thr Asp Tyr His
645 650 655
Ile Asp Gln Val Ser Asn Leu Val Thr Tyr Leu Ser Asp Glu Phe Cys
660 665 670
Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg
675 680 685
Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Ser Asn Phe Lys Asp Ile
690 695 700
Asn Arg Gln Pro Glu Arg Gly Trp Gly Gly Ser Thr Gly Ile Thr Ile
705 710 715 720
Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Ser Gly
725 730 735
Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu
740 745 750
Ser Lys Leu Lys Ala Phe Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu
755 760 765
Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His
770 775 780
Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala
785 790 795 800
Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His
805 810 815
Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys
820 825 830
Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys
835 840 845
Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys
850 855 860
Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu
865 870 875 880
Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys
885 890 895
Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val
900 905 910
Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln
915 920 925
Tyr Asp Gln Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala
930 935 940
Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser
945 950 955 960
Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg
965 970 975
Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn
980 985 990
Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val
995 1000 1005
Asp Val Glu Glu Gln Asn Asn Gln Arg Ser Val Leu Val Val Pro
1010 1015 1020
Glu Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly
1025 1030 1035
Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly
1040 1045 1050
Glu Gly Cys Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu
1055 1060 1065
Leu Lys Phe Ser Asn Cys Val Glu Glu Glu Ile Tyr Pro Asn Asn
1070 1075 1080
Thr Val Thr Cys Asn Asp Tyr Thr Val Asn Gln Glu Glu Tyr Gly
1085 1090 1095
Gly Ala Tyr Thr Ser Arg Asn Arg Gly Tyr Asn Glu Ala Pro Ser
1100 1105 1110
Val Pro Ala Asp Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr
1115 1120 1125
Asp Gly Arg Arg Glu Asn Pro Cys Glu Phe Asn Arg Gly Tyr Arg
1130 1135 1140
Asp Tyr Thr Pro Leu Pro Val Gly Tyr Val Thr Lys Glu Leu Glu
1145 1150 1155
Tyr Phe Pro Glu Thr Asp Lys Val Trp Ile Glu Ile Gly Glu Thr
1160 1165 1170
Glu Gly Thr Phe Ile Val Asp Ser Val Glu Leu Leu Leu Met Glu
1175 1180 1185
Glu
<210> 5
<211> 3672
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC860.
<400> 5
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccaacggta 60
tcgaatcctt ccacgcaaat gaatctatca ccagatgctc gtattgaaga tagcttgtgt 120
gtagccgagg tgaacaatat tgatccattt gttagcgcat caacagtcca aacgggtata 180
aacatagctg gtagaatatt gggcgtatta ggtgtgccgt ttgctggaca actagctagt 240
ttttatagtt ttcttgttgg ggaattatgg cctagtggca gagatccatg ggaaattttc 300
ctggaacatg tagaacaact tataagacaa caagtaacag aaaatactag gaatacggct 360
attgctcgat tagaaggtct aggaagaggc tatagatctt accagcaggc tcttgaaact 420
tggttagata accgaaatga tgcaagatca agaagcatta ttcttgagcg ctatgttgct 480
ttagaacttg acattactac tgctataccg cttttcagaa tacgaaatga agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agacgcatcc 600
ctttttggta gtgaatgggg gatggcatct tccgatgtta accaatatta ccaagaacaa 660
atcagatata cagaggaata ttctaaccat tgcgtacaat ggtataatac agggctaaat 720
aacttaagag ggacaaatgc tgaaagttgg ttgcggtata atcaattccg tagagaccta 780
acgttagggg tattagattt agtagcccta ttcccaagct atgatactcg cacttatcca 840
atcaatacga gtgctcagtt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccattttcag gcctccgcat ctacttgatt ttccagaaca acttacaatt 1020
tacagtgcat caagccgttg gagtagcact caacatatga attattgggt gggacatagg 1080
cttaacttcc gcccaatagg agggacatta aatacctcaa cacaaggact tactaataat 1140
acttcaatta atcctgtaac attacagttt acgtctcgag acgtttatag aacagaatca 1200
aatgcaggga caaatatact atttactact cctgtgaatg gagtaccttg ggctagattt 1260
aattttataa accctcagaa tatttatgaa agaggcgcca ctacctacag tcaaccgtat 1320
cagggagttg ggattcaatt atttgattca gaaactgaat taccaccaga aacaacagaa 1380
cgaccaaatt atgaatcata tagtcataga ttatctcata taggactaat cataggaaac 1440
actttgagag caccagtcta ttcttggacg catcgtagtg cagatcgtac gaatacgatt 1500
ggaccaaata gaattaatca aataccttta gtgaaaggat ttagagtttg ggggggcacc 1560
tctgtcatta caggaccagg atttacagga ggggatatcc ttcgaagaaa tacctttggt 1620
gattttgtat ctctacaagt caatattaat tcaccaatta cccaaagata ccgtttaaga 1680
tttcgttacg cttccagtag ggatgcacga gttatagtat taacaggagc ggcatccaca 1740
ggagtgggag gccaagttag tgtaaatatg cctcttcaga aaactatgga aataggggag 1800
aacttaacat ctagaacatt tagatatacc gattttagta atcctttttc atttagagct 1860
aatccagata taattgggat aagtgaacaa cctctatttg gtgcaggttc tattagtagc 1920
ggtgaacttt atatagataa aattgaaatt attctagcag atgcaacatt tgaagcagaa 1980
tctgatttag aaagagcgca gaaggcggtg aatgcgctgt ttacgtctac aaaccaacta 2040
gggctaaaaa caaatgtaac ggattatcat attgatcaag tgtccaattt agttacgtat 2100
ttatcggatg aattttgtct ggatgaaaag cgagaattgt ccgagaaagt caaacatgcg 2160
aagcgactca gtgatgaacg caatttactc caagattcaa atttcaaaga cattaatagg 2220
caaccagaac gtgggtgggg cggaagtaca gggattacca tccaaggagg ggatgacgta 2280
tttaaagaaa attacgtcac actatcaggt acctttgatg agtgctatcc aacatatttg 2340
tatcaaaaaa tcgatgaatc aaaattaaaa gcctttaccc gttatcaatt aagagggtat 2400
atcgaagata gtcaagactt agaaatctat ttaattcgct acaatgcaaa acatgaaaca 2460
gtaaatgtgc caggtacggg ttccttatgg ccgctttcag cccaaagtcc aatcggaaag 2520
tgtggagagc cgaatcgatg cgcgccacac cttgaatgga atcctgactt agattgttcg 2580
tgtagggatg gagaaaagtg tgcccatcat tcgcatcatt tctccttaga cattgatgta 2640
ggatgtacag acttaaatga ggacctaggt gtatgggtga tctttaagat taagacgcaa 2700
gatgggcacg caagactagg gaatctagag tttctcgaag agaaaccatt agtaggagaa 2760
gcgctagctc gtgtgaaaag agcggagaaa aaatggagag acaaacgtga aaaattggaa 2820
tgggaaacaa atatcgttta taaagaggca aaagaatctg tagatgcttt atttgtaaac 2880
tctcaatatg atcaattaca agcggatacg aatattgcca tgattcatgc ggcagataaa 2940
cgtgttcata gcattcgaga agcttatctg cctgagctgt ctgtgattcc gggtgtcaat 3000
gcggctattt ttgaagaatt agaagggcgt attttcactg cattctccct atatgatgcg 3060
agaaatgtca ttaaaaatgg tgattttaat aatggcttat cctgctggaa cgtgaaaggg 3120
catgtagatg tagaagaaca aaacaaccaa cgttcggtcc ttgttgttcc ggaatgggaa 3180
gcagaagtgt cacaagaagt tcgtgtctgt ccgggtcgtg gctatatcct tcgtgtcaca 3240
gcgtacaagg agggatatgg agaaggttgc gtaaccattc atgagatcga gaacaataca 3300
gacgaactga agtttagcaa ctgcgtagaa gaggaaatct atccaaataa cacggtaacg 3360
tgtaatgatt atactgtaaa tcaagaagaa tacggaggtg cgtacacttc tcgtaatcga 3420
ggatataacg aagctccttc cgtaccagct gattatgcgt cagtctatga agaaaaatcg 3480
tatacagatg gacgaagaga gaatccttgt gaatttaaca gagggtatag ggattacacg 3540
ccactaccag ttggttatgt gacaaaagaa ttagaatact tcccagaaac cgataaggta 3600
tggattgaga ttggagaaac ggaaggaaca tttatcgtgg acagcgtgga attactcctt 3660
atggaggaat ag 3672
<210> 6
<211> 3672
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC860.
<400> 6
atgaccagca accggaagaa cgagaacgag atcatcaacg ccctgagcat cccgaccgtg 60
agcaacccta gcacccagat gaacctgagc cctgacgctc gcatcgagga ctccctctgc 120
gtggctgagg tgaacaacat cgacccgttc gtgtccgcct ccaccgtgca gaccggcatc 180
aacatcgcgg gccgcatcct cggcgtgctc ggcgtgccct ttgcgggcca gctcgcctcc 240
ttctactcct tcctcgtggg agagctgtgg ccctccggcc gcgacccgtg ggagatcttc 300
ctggagcacg tggagcagct catccgccag caagtcaccg agaacacccg caacaccgcc 360
atcgcccgcc tggagggcct gggccgtggc taccgctcct accagcaagc cctggagacc 420
tggctcgaca accgcaacga cgcccgctcc cgctccatca tcctggagcg ctacgtcgcc 480
ctggaactgg acatcaccac tgccatccca ctcttccgca tcaggaacga ggaggtgcct 540
ctgctgatgg tgtacgccca ggctgcgaac ctgcacctgc tgctgctgcg cgacgcaagc 600
ctgtttggct ccgagtgggg tatggcaagc tccgacgtca accagtacta ccaggagcag 660
atccgctaca ccgaggagta cagcaaccac tgcgtccagt ggtacaacac cggtctgaac 720
aatctcagag ggaccaacgc tgagagctgg ctgcgctaca accagttccg gcgggatctg 780
accctaggtg tcctggatct ggtcgctctg ttcccgagct acgataccag gacgtaccct 840
atcaacacct ctgctcagct taccagggag atctacactg atcctatcgg taggactaac 900
gctcctagtg gtttcgccag cactaactgg ttcaacaaca acgcgcctag tttctctgcc 960
atcgaggcgg cgatcttccg gcctcctcac ctcctcgact tcccggagca gcttactatc 1020
tactctgcgt cttcgcggtg gtcttcgact cagcacatga actactgggt tggtcaccgg 1080
cttaacttcc gcccgattgg aggaactctt aacaccagta cgcaaggtct tacgaacaac 1140
acttccatca acccggttac gttgcagttc acgtctcggg acgtttaccg gacggagtcg 1200
aatgctggga cgaacatcct gttcacgaca ccggtgaatg gtgttccgtg ggcacgtttc 1260
aacttcatca acccgcagaa catctacgag cgtggagcaa cgacatactc gcaaccatac 1320
caaggcgttg gcatccaact gtttgactcg gagacggaac tgccaccaga gacgacagaa 1380
cgtccgaatt acgagtcata ctcacacaga ctatcacaca ttggactcat tatcggaaac 1440
acactgagag caccagtgta ctcatggaca catcggtcag cagatcgtac gaacaccatc 1500
ggacccaatc ggatcaacca gatcccgctc gtgaagggct tccgcgtgtg gggcggcacc 1560
tccgtcatca ccggtccggg cttcaccggc ggcgacatcc tccgccgcaa caccttcggc 1620
gacttcgtgt cactccaagt gaacatcaac agcccgatca cccagcgcta tcgcctccgc 1680
ttccgctacg cctcctcccg cgacgctaga gtgatcgtgc tcaccggagc ggcgtccaca 1740
ggcgtaggcg gccaagtgtc tgtgaacatg ccgctccaga agactatgga gattggtgag 1800
aacctcacct ctcgcacctt ccgctacacc gacttctcca atccgttctc cttcagagcc 1860
aacccagaca tcatcggcat ctccgagcag cctctctttg gcgctggctc catctcctcc 1920
ggcgagctgt acatcgacaa gattgagatc atccttgccg acgccacctt cgaagctgag 1980
tccgatctcg agcgcgccca gaaggccgtg aacgccctct tcactagcac taaccagctc 2040
ggcctcaaga ctaacgtgac cgactaccac attgaccaag tgagcaacct agtgacctac 2100
cttagcgacg agttctgcct tgacgagaag cgtgagctga gcgagaaggt gaagcacgcc 2160
aagcgcctct ccgacgagcg caacctcctc caggactcca acttcaagga catcaaccgc 2220
cagcccgagc gcggctgggg cggtagcacc ggcatcacca tccagggcgg tgacgatgtg 2280
ttcaaggaga actacgtgac cctctccggc accttcgacg agtgctaccc gacctacctc 2340
taccagaaga tcgacgagtc caagctcaag gcgttcaccc gctaccagct tcgcggctac 2400
atcgaggact cccaggatct ggagatctac ctcatccgct acaacgccaa gcacgagacc 2460
gtgaacgtgc ccggcaccgg ctccctctgg ccgctctccg cccagagccc tatcggcaag 2520
tgcggcgagc ccaaccgctg cgcgcctcac ctggagtgga accctgacct cgactgctcc 2580
tgccgcgacg gcgagaagtg cgcccaccat agccaccact tctctctcga catcgacgtg 2640
ggctgcaccg acctcaacga ggatctgggc gtgtgggtga tcttcaagat caagacccag 2700
gacggccacg ccaggctggg caacctggag ttcctggagg agaagcctct ggtgggtgag 2760
gccctggcca gggtcaagag ggctgagaag aaatggaggg acaagaggga gaagctggag 2820
tgggagacca acatcgtgta caaggaggct aaggagtccg tggacgctct gttcgtcaac 2880
tctcagtacg atcagctcca ggctgacacc aacatcgcta tgatccacgc tgcggataag 2940
agggtccact ctatcaggga ggcttacctg cctgagcttt ctgtcatccc tggtgtcaac 3000
gcggcaatct tcgaggaact tgagggccgc atcttcactg cgttctcgct ttacgatgcg 3060
cggaacgtca ttaagaacgg tgacttcaac aatggtcttt cgtgctggaa cgtcaagggt 3120
catgtcgatg tcgaggaaca gaacaaccag cggtcggtcc ttgtcgttcc cgagtgggag 3180
gccgaggtct cgcaagaggt ccgggtctgc cctgggcgcg ggtacattct tcgtgtcact 3240
gcgtacaagg agggctacgg cgagggctgc gttactattc atgagattga gaacaatacg 3300
gatgagctta agtttagtaa ctgtgttgag gaggagatct acccgaacaa tacggttacg 3360
tgcaatgatt acacggtgaa ccaggaggaa tacggcggag catacacctc acgtaataga 3420
gggtacaatg aggcaccgtc agttccggca gattatgcct cagtttatga ggagaagtcc 3480
tacacggatg gaagacgcga gaatccatgt gagtttaata gaggataccg agactacaca 3540
ccactcccag ttggatacgt tacaaaggag ttggaatact tcccagaaac agataaagtt 3600
tggatagaga tcggagaaac agaaggaacc ttcatcgtgg acagtgtaga actgctgctg 3660
atggaagaat ga 3672
<210> 7
<211> 1223
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein TIC860.
<400> 7
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Thr Val Ser Asn Pro Ser Thr Gln Met Asn Leu Ser Pro Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Val Ala Glu Val Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Leu Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Ser Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asn Thr Ala Ile Ala Arg Leu Glu Gly Leu Gly
115 120 125
Arg Gly Tyr Arg Ser Tyr Gln Gln Ala Leu Glu Thr Trp Leu Asp Asn
130 135 140
Arg Asn Asp Ala Arg Ser Arg Ser Ile Ile Leu Glu Arg Tyr Val Ala
145 150 155 160
Leu Glu Leu Asp Ile Thr Thr Ala Ile Pro Leu Phe Arg Ile Arg Asn
165 170 175
Glu Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Trp Gly Met
195 200 205
Ala Ser Ser Asp Val Asn Gln Tyr Tyr Gln Glu Gln Ile Arg Tyr Thr
210 215 220
Glu Glu Tyr Ser Asn His Cys Val Gln Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Thr Tyr Pro Ile Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Ile Phe Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Tyr Ser Ala Ser Ser Arg Trp Ser Ser Thr Gln His
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Asn Phe Arg Pro Ile Gly Gly
355 360 365
Thr Leu Asn Thr Ser Thr Gln Gly Leu Thr Asn Asn Thr Ser Ile Asn
370 375 380
Pro Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser
385 390 395 400
Asn Ala Gly Thr Asn Ile Leu Phe Thr Thr Pro Val Asn Gly Val Pro
405 410 415
Trp Ala Arg Phe Asn Phe Ile Asn Pro Gln Asn Ile Tyr Glu Arg Gly
420 425 430
Ala Thr Thr Tyr Ser Gln Pro Tyr Gln Gly Val Gly Ile Gln Leu Phe
435 440 445
Asp Ser Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr
450 455 460
Glu Ser Tyr Ser His Arg Leu Ser His Ile Gly Leu Ile Ile Gly Asn
465 470 475 480
Thr Leu Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg
485 490 495
Thr Asn Thr Ile Gly Pro Asn Arg Ile Asn Gln Ile Pro Leu Val Lys
500 505 510
Gly Phe Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe
515 520 525
Thr Gly Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser
530 535 540
Leu Gln Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg
545 550 555 560
Phe Arg Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly
565 570 575
Ala Ala Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu
580 585 590
Gln Lys Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg
595 600 605
Tyr Thr Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile
610 615 620
Ile Gly Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser
625 630 635 640
Gly Glu Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr
645 650 655
Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala
660 665 670
Leu Phe Thr Ser Thr Asn Gln Leu Gly Leu Lys Thr Asn Val Thr Asp
675 680 685
Tyr His Ile Asp Gln Val Ser Asn Leu Val Thr Tyr Leu Ser Asp Glu
690 695 700
Phe Cys Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys His Ala
705 710 715 720
Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Ser Asn Phe Lys
725 730 735
Asp Ile Asn Arg Gln Pro Glu Arg Gly Trp Gly Gly Ser Thr Gly Ile
740 745 750
Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu
755 760 765
Ser Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile
770 775 780
Asp Glu Ser Lys Leu Lys Ala Phe Thr Arg Tyr Gln Leu Arg Gly Tyr
785 790 795 800
Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala
805 810 815
Lys His Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu
820 825 830
Ser Ala Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala
835 840 845
Pro His Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly
850 855 860
Glu Lys Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val
865 870 875 880
Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys
885 890 895
Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu
900 905 910
Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala
915 920 925
Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn
930 935 940
Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn
945 950 955 960
Ser Gln Tyr Asp Gln Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His
965 970 975
Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu
980 985 990
Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu
995 1000 1005
Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val
1010 1015 1020
Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val
1025 1030 1035
Lys Gly His Val Asp Val Glu Glu Gln Asn Asn Gln Arg Ser Val
1040 1045 1050
Leu Val Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val Arg
1055 1060 1065
Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys
1070 1075 1080
Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile Glu Asn
1085 1090 1095
Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val Glu Glu Glu Ile
1100 1105 1110
Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp Tyr Thr Val Asn Gln
1115 1120 1125
Glu Glu Tyr Gly Gly Ala Tyr Thr Ser Arg Asn Arg Gly Tyr Asn
1130 1135 1140
Glu Ala Pro Ser Val Pro Ala Asp Tyr Ala Ser Val Tyr Glu Glu
1145 1150 1155
Lys Ser Tyr Thr Asp Gly Arg Arg Glu Asn Pro Cys Glu Phe Asn
1160 1165 1170
Arg Gly Tyr Arg Asp Tyr Thr Pro Leu Pro Val Gly Tyr Val Thr
1175 1180 1185
Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp Lys Val Trp Ile Glu
1190 1195 1200
Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp Ser Val Glu Leu
1205 1210 1215
Leu Leu Met Glu Glu
1220
<210> 8
<211> 3564
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC867.
<400> 8
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta cacaaatacc attggtaaag gcgcataccc tccaatcggg taccactgta 1560
gtaaaagggc cagggtttac aggaggggat atcctccgtc gaacaagtgg aggaccattt 1620
gcttttagta atgttaatct agattttaac ttgtcacaaa ggtatcgtgc tagaattcgt 1680
tatgcctcta ctactaacct aagaatttac gtaacggttg caggtgaacg aatttttgct 1740
ggtcaatttg acaaaactat ggatgctggt gccccattaa cattccaatc ttttagttac 1800
gcaactatta atacagcttt tacattccca gaaagatcga gcagcttgac tgtaggtgcc 1860
gatacgttta gttcaggtaa tgaagtttat gtagatagat ttgaattaat cccagttact 1920
gcaaccttcg aggcagaatc tgatttagaa agagcacaaa aggcggtgaa tgagctgttt 1980
acttcttcca atcaaatcgg gttaaaaaca gatgtgacgg attatcatat tgatcaagta 2040
tccaatttag ttgagtgttt atctgatgaa ttttgtctgg atgaaaaaaa agaattgtcc 2100
gagaaagtca aacatgcgaa gcgacttagt gatgagcgga atttacttca agatccaaac 2160
tttagaggga tcaatagaca actagaccgt ggctggagag gaagtacgga tattaccatc 2220
caaggaggcg atgacgtatt caaagagaat tacgttacgc tattgggtac ctttgatgag 2280
tgctatccaa cgtatttata tcaaaaaata gatgagtcga aattaaaagc ctatacccgt 2340
taccaattaa gagggtatat cgaagatagt caagacttag aaatctattt aattcgctac 2400
aatgccaaac acgaaacagt aaatgtgcca ggtacgggtt ccttatggcc gctttcagcc 2460
ccaagtccaa tcggaaaatg tgcccatcat tcccatcatt tctccttgga cattgatgtt 2520
ggatgtacag acttaaatga ggacttaggt gtatgggtga tattcaagat taagacgcaa 2580
gatggccatg caagactagg aaatctagaa tttctcgaag agaaaccatt agtaggagaa 2640
gcactagctc gtgtgaaaag agcggagaaa aaatggagag acaaacgtga aaaattggaa 2700
tgggaaacaa atattgttta taaagaggca aaagaatctg tagatgcttt atttgtaaac 2760
tctcaatatg atagattaca agcggatacc aacatcgcga tgattcatgc ggcagataaa 2820
cgcgttcata gcattcgaga agcttatctg cctgagctgt ctgtgattcc gggtgtcaat 2880
gcggctattt ttgaagaatt agaagggcgt attttcactg cattctccct atatgatgcg 2940
agaaatgtca ttaaaaatgg tgattttaat aatggcttat cctgctggaa cgtgaaaggg 3000
catgtagatg tagaagaaca aaacaaccac cgttcggtcc ttgttgttcc ggaatgggaa 3060
gcagaagtgt cacaagaagt tcgtgtctgt ccgggtcgtg gctatatcct tcgtgtcaca 3120
gcgtacaagg agggatatgg agaaggttgc gtaaccattc atgagatcga gaacaataca 3180
gacgaactga agtttagcaa ctgtgtagaa gaggaagtat atccaaacaa cacggtaacg 3240
tgtaatgatt atactgcgac tcaagaagaa tatgagggta cgtacacttc tcgtaatcga 3300
ggatatgacg gagcctatga aagcaattct tctgtaccag ctgattatgc atcagcctat 3360
gaagaaaaag catatacaga tggacgaaga gacaatcctt gtgaatctaa cagaggatat 3420
ggggattaca caccactacc agctggctat gtgacaaaag aattagagta cttcccagaa 3480
accgataagg tatggattga gatcggagaa acggaaggaa cattcatcgt ggacagcgtg 3540
gaattacttc ttatggagga atag 3564
<210> 9
<211> 3564
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867.
<400> 9
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccttcg aagctgagtc ggacctggag cgtgcacaga aggcagtcaa cgagctgttc 1980
acctctagca accagatcgg cctcaagacc gacgtcacag actaccacat cgaccaagtg 2040
tccaacctgg tcgagtgcct tagcgacgag ttctgcctag acgagaagaa ggagctgtcg 2100
gagaaggtca aacacgccaa gcgtctgagc gatgagcgca acctgctcca agaccctaac 2160
ttccgtggca tcaacaggca gcttgaccgt ggctggcgcg gctcgacgga catcacgatc 2220
cagggtggcg acgacgtatt caaggagaat tacgtgacct tgcttgggac gtttgacgag 2280
tgctatccca cctacctcta ccagaagatt gatgaatcga aattgaaggc gtacacgaga 2340
taccagctcc gtggctacat cgaggacagc caggacttgg agatctacct catacgctac 2400
aacgctaaac atgagaccgt gaacgtccct gggacgggca gtctgtggcc actctctgct 2460
cctagcccta tcggcaagtg cgctcaccac tcgcaccact tcagccttga catcgacgtg 2520
ggatgtactg acctcaacga agacctgggc gtctgggtta tcttcaagat caagacccag 2580
gacggccacg cccgactcgg caacctggag ttcctggagg agaaaccact ggtgggcgag 2640
gcgctcgccc gcgtgaagcg tgccgagaag aagtggcggg acaagaggga gaagctagaa 2700
tgggagacga acatcgtgta caaggaggcc aaggaaagcg tcgatgccct gttcgtgaac 2760
tcacagtacg accgtctcca ggcggacacg aacatcgcca tgatccacgc ggctgacaag 2820
cgcgtccact ccatccgcga ggcgtactta ccggagctgt cggtgatccc aggcgtaaac 2880
gcggcgatct tcgaggagct agagggacgc atcttcacag cgttcagcct gtacgacgca 2940
cgcaacgtca tcaagaacgg cgatttcaac aacggactgt cctgctggaa cgtgaagggc 3000
cacgtcgatg tcgaggaaca gaacaaccac cgctctgtcc tggtggtccc agagtgggag 3060
gccgaggtct cccaggaggt ccgcgtgtgc cctgggcgtg gctacatcct ccgtgtgaca 3120
gcctacaagg agggctacgg tgagggctgc gtcaccattc acgagatcga gaacaacact 3180
gacgaactca agttctcgaa ttgcgtggag gaggaggtgt acccgaacaa tacggtgacg 3240
tgcaacgact acacggcaac ccaagaggag tacgagggca cctacaccag taggaaccgt 3300
ggctacgacg gtgcctacga gtcgaactcc agcgtccctg cggactacgc cagcgcgtac 3360
gaggagaagg cttacaccga cggacgccgg gacaacccat gcgagagcaa ccgtggctac 3420
ggcgactaca ctcctctccc ggccggatac gtcacaaagg agctggagta tttcccagag 3480
acggacaagg tgtggatcga aatcggagag acagagggaa ccttcatcgt ggacagcgtg 3540
gagctgctcc tcatggagga gtga 3564
<210> 10
<211> 1187
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein TIC867.
<400> 10
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val
645 650 655
Asn Glu Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val
660 665 670
Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser
675 680 685
Asp Glu Phe Cys Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys
690 695 700
His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn
705 710 715 720
Phe Arg Gly Ile Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr
725 730 735
Asp Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val
740 745 750
Thr Leu Leu Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln
755 760 765
Lys Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg
770 775 780
Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr
785 790 795 800
Asn Ala Lys His Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp
805 810 815
Pro Leu Ser Ala Pro Ser Pro Ile Gly Lys Cys Ala His His Ser His
820 825 830
His Phe Ser Leu Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp
835 840 845
Leu Gly Val Trp Val Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala
850 855 860
Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu Lys Pro Leu Val Gly Glu
865 870 875 880
Ala Leu Ala Arg Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg
885 890 895
Glu Lys Leu Glu Trp Glu Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu
900 905 910
Ser Val Asp Ala Leu Phe Val Asn Ser Gln Tyr Asp Arg Leu Gln Ala
915 920 925
Asp Thr Asn Ile Ala Met Ile His Ala Ala Asp Lys Arg Val His Ser
930 935 940
Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser Val Ile Pro Gly Val Asn
945 950 955 960
Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg Ile Phe Thr Ala Phe Ser
965 970 975
Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly
980 985 990
Leu Ser Cys Trp Asn Val Lys Gly His Val Asp Val Glu Glu Gln Asn
995 1000 1005
Asn His Arg Ser Val Leu Val Val Pro Glu Trp Glu Ala Glu Val
1010 1015 1020
Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg
1025 1030 1035
Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile
1040 1045 1050
His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys
1055 1060 1065
Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp
1070 1075 1080
Tyr Thr Ala Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg
1085 1090 1095
Asn Arg Gly Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro
1100 1105 1110
Ala Asp Tyr Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp Gly
1115 1120 1125
Arg Arg Asp Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr
1130 1135 1140
Thr Pro Leu Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe
1145 1150 1155
Pro Glu Thr Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly
1160 1165 1170
Thr Phe Ile Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1175 1180 1185
<210> 11
<211> 3642
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC867_20.
<400> 11
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta cacaaatacc attggtaaag gcgcataccc tccaatcggg taccactgta 1560
gtaaaagggc cagggtttac aggaggggat atcctccgtc gaacaagtgg aggaccattt 1620
gcttttagta atgttaatct agattttaac ttgtcacaaa ggtatcgtgc tagaattcgt 1680
tatgcctcta ctactaacct aagaatttac gtaacggttg caggtgaacg aatttttgct 1740
ggtcaatttg acaaaactat ggatgctggt gccccattaa cattccaatc ttttagttac 1800
gcaactatta atacagcttt tacattccca gaaagatcga gcagcttgac tgtaggtgcc 1860
gatacgttta gttcaggtaa tgaagtttat gtagatagat ttgaattaat cccagttact 1920
gcaacctttg aggcagaata tgatttagaa agagcgcaaa aggtggtgaa tgccctgttt 1980
acgtctacaa accaactagg gctaaaaaca gatgtgacgg attatcatat tgatcaggta 2040
tccaatctag ttgcgtgttt atcggatgaa ttttgtctgg atgaaaagag agaattgtcc 2100
gagaaagtta aacatgcaaa gcgactcagt gatgagcgga atttacttca agatccaaac 2160
ttcagaggga tcaataggca accagaccgt ggctggagag gaagtacgga tattactatc 2220
caaggaggag atgacgtatt caaagagaat tacgttacgc taccgggtac ctttgatgag 2280
tgctatccaa cgtatttata tcaaaaaata gatgagtcga aattaaaagc ctatacccgt 2340
tatcaattaa gagggtatat cgaagatagt caagacttag aaatctattt aattcgttac 2400
aatgcaaaac acgaaatagt aaatgtacca ggtacaggaa gtttatggcc tctttctgta 2460
gaaaatcaaa ttggaccttg tggagaaccg aatcgatgcg cgccacacct tgaatggaat 2520
cctgatttac actgttcctg cagagacggg gaaaaatgtg cacatcattc tcatcatttc 2580
tctttggaca ttgatgttgg atgtacagac ttaaatgagg acttaggtgt atgggtgata 2640
ttcaagatta agacgcaaga tggccacgca cgactaggga atctagagtt tctcgaagag 2700
aaaccattat taggagaagc actagctcgt gtgaaaagag cggagaaaaa atggagagac 2760
aaacgcgaaa cattacaatt ggaaacaact atcgtttata aagaggcaaa agaatctgta 2820
gatgctttat ttgtaaactc tcaatatgat agattacaag cggatacgaa catcgcgatg 2880
attcatgcgg cagataaacg cgttcataga attcgagaag cgtatctgcc ggagctgtct 2940
gtgattccgg gtgtcaatgc ggctattttt gaagaattag aagagcgtat tttcactgca 3000
ttttccctat atgatgcgag aaatattatt aaaaatggcg atttcaataa tggcttatta 3060
tgctggaacg tgaaagggca tgtagaggta gaagaacaaa acaatcaccg ttcagtcctg 3120
gttatcccag aatgggaggc agaagtgtca caagaggttc gtgtctgtcc aggtcgtggc 3180
tatatccttc gtgttacagc gtacaaagag ggatatggag aaggttgcgt aacgatccat 3240
gagatcgaga acaatacaga cgaactgaaa ttcaacaact gtgtagaaga ggaagtatat 3300
ccaaacaaca cggtaacgtg tattaattat actgcgactc aagaagaata tgagggtacg 3360
tacacttctc gtaatcgagg atatgacgaa gcctatggta ataacccttc cgtaccagct 3420
gattatgcgt cagtctatga agaaaaatcg tatacagata gacgaagaga gaatccttgt 3480
gaatctaaca gaggatatgg agattacaca ccactaccag ctggttatgt aacaaaggaa 3540
ttagagtact tcccagagac cgataaggta tggattgaga ttggagaaac agaaggaaca 3600
ttcatcgtgg acagcgtgga attactcctt atggaggaat ag 3642
<210> 12
<211> 3642
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867_20.
<400> 12
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccttcg aggccgagta cgaccttgag cgcgcccaga aggtggtgaa cgccctcttc 1980
actagcacta accagctagg cctgaagact gacgtgaccg actaccacat cgaccaagtg 2040
agcaacctag tggcctgcct ctccgacgag ttctgcctcg acgagaagcg cgagctgtcc 2100
gagaaggtga agcacgccaa gcgcctctcc gacgagcgca acctgctcca ggaccccaac 2160
ttcaggggca tcaacaggca gcccgaccgc ggctggcgcg gctccaccga catcaccatc 2220
cagggcggtg acgacgtatt caaggagaac tacgttaccc tccccggcac cttcgacgag 2280
tgttacccca cctacctcta ccagaagatc gacgagtcca agctgaaggc ctacacccgc 2340
taccagctcc gcggctacat cgaggactcc caggacctgg aaatctacct catccgctac 2400
aacgccaagc acgagatcgt gaacgtgcct ggcaccggca gcctctggcc tctcagcgtg 2460
gagaaccaga tcggcccttg cggcgagcct aaccgctgcg cccctcacct cgagtggaac 2520
cctgacctcc actgctcgtg cagggacggc gagaagtgcg cccaccatag ccaccacttc 2580
tctctggaca tcgacgtggg ctgcaccgac ctgaacgagg acctgggcgt gtgggttatc 2640
ttcaagatca agacccagga cggtcacgcc aggctgggta acctggagtt ccttgaggaa 2700
aagcctctgc tgggtgaggc cctggccagg gtcaagaggg ctgagaagaa atggagggat 2760
aagagggaga ccctgcagct ggagaccact atcgtctaca aggaggctaa ggagtctgtc 2820
gatgctctgt tcgtcaactc tcagtacgat agactgcaag ctgataccaa catcgctatg 2880
atccacgctg cggataagcg ggtccaccgg atccgggagg cttaccttcc ggagctttct 2940
gtcatcccgg gtgtcaacgc tgcgatcttc gaggaacttg aggaacggat cttcactgcg 3000
tttagtcttt acgatgcgcg gaacatcatc aagaacgggg acttcaacaa tggtctgctg 3060
tgctggaacg tcaagggtca tgtcgaggtc gaggaacaaa acaatcatcg tagtgtcctt 3120
gtcattcctg agtgggaggc ggaggtctct caagaggtcc gtgtttgccc ggggcgtggg 3180
tacattcttc gtgttactgc gtacaaggag gggtacgggg aggggtgcgt tactattcat 3240
gagattgaga acaatactga tgagcttaag ttcaacaatt gtgttgagga ggaggtttac 3300
ccgaacaata ctgttacgtg catcaactac acggcaacgc aagaggaata cgaggggacg 3360
tacacctcgc gtaatagagg gtatgatgag gcgtacggaa acaacccgtc ggttccagca 3420
gattatgcct cggtttatga ggagaagtcg tacacggata gacgacgcga gaatccatgt 3480
gagtcaaatc gaggatacgg agattacaca ccattaccag caggatacgt tacaaaggag 3540
ttggaatact tcccggaaac agataaagtt tggattgaaa tcggagaaac agaaggaaca 3600
ttcatcgtcg actcagtaga attgttgttg atggaagaat ga 3642
<210> 13
<211> 1213
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC867_20.
<400> 13
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg Ala Gln Lys Val Val
645 650 655
Asn Ala Leu Phe Thr Ser Thr Asn Gln Leu Gly Leu Lys Thr Asp Val
660 665 670
Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Ala Cys Leu Ser
675 680 685
Asp Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys
690 695 700
His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn
705 710 715 720
Phe Arg Gly Ile Asn Arg Gln Pro Asp Arg Gly Trp Arg Gly Ser Thr
725 730 735
Asp Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val
740 745 750
Thr Leu Pro Gly Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln
755 760 765
Lys Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg
770 775 780
Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr
785 790 795 800
Asn Ala Lys His Glu Ile Val Asn Val Pro Gly Thr Gly Ser Leu Trp
805 810 815
Pro Leu Ser Val Glu Asn Gln Ile Gly Pro Cys Gly Glu Pro Asn Arg
820 825 830
Cys Ala Pro His Leu Glu Trp Asn Pro Asp Leu His Cys Ser Cys Arg
835 840 845
Asp Gly Glu Lys Cys Ala His His Ser His His Phe Ser Leu Asp Ile
850 855 860
Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile
865 870 875 880
Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu
885 890 895
Phe Leu Glu Glu Lys Pro Leu Leu Gly Glu Ala Leu Ala Arg Val Lys
900 905 910
Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Thr Leu Gln Leu Glu
915 920 925
Thr Thr Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe
930 935 940
Val Asn Ser Gln Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Ala Met
945 950 955 960
Ile His Ala Ala Asp Lys Arg Val His Arg Ile Arg Glu Ala Tyr Leu
965 970 975
Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu
980 985 990
Leu Glu Glu Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn
995 1000 1005
Ile Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Leu Cys Trp Asn
1010 1015 1020
Val Lys Gly His Val Glu Val Glu Glu Gln Asn Asn His Arg Ser
1025 1030 1035
Val Leu Val Ile Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val
1040 1045 1050
Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr
1055 1060 1065
Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile Glu
1070 1075 1080
Asn Asn Thr Asp Glu Leu Lys Phe Asn Asn Cys Val Glu Glu Glu
1085 1090 1095
Val Tyr Pro Asn Asn Thr Val Thr Cys Ile Asn Tyr Thr Ala Thr
1100 1105 1110
Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Arg Gly Tyr
1115 1120 1125
Asp Glu Ala Tyr Gly Asn Asn Pro Ser Val Pro Ala Asp Tyr Ala
1130 1135 1140
Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Arg Arg Arg Glu Asn
1145 1150 1155
Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu Pro
1160 1165 1170
Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp
1175 1180 1185
Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val
1190 1195 1200
Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1205 1210
<210> 14
<211> 3690
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC867_21.
<400> 14
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta cacaaatacc attggtaaag gcgcataccc tccaatcggg taccactgta 1560
gtaaaagggc cagggtttac aggaggggat atcctccgtc gaacaagtgg aggaccattt 1620
gcttttagta atgttaatct agattttaac ttgtcacaaa ggtatcgtgc tagaattcgt 1680
tatgcctcta ctactaacct aagaatttac gtaacggttg caggtgaacg aatttttgct 1740
ggtcaatttg acaaaactat ggatgctggt gccccattaa cattccaatc ttttagttac 1800
gcaactatta atacagcttt tacattccca gaaagatcga gcagcttgac tgtaggtgcc 1860
gatacgttta gttcaggtaa tgaagtttat gtagatagat ttgaattaat cccagttact 1920
gcaaccggaa cgacaaccta tgagtatgaa gagaagcaga atctagaaaa agcgcagaaa 1980
gcgttgaacg ctttgtttac ggatggcacg aatggctatc tacaaatgga tgccactgat 2040
tatgatatca atcaaactgc aaacttaata gaatgtgtat cagatgaatt gtatgcaaaa 2100
gaaaagatag ttttattaga tgaagtcaaa tatgcgaagc ggcttagcat atcacgtaac 2160
ctacttttga acgatgattt agaattttca gatggatttg gagaaaacgg atggacgaca 2220
agtgataata tttcaatcca ggcggataat ccccttttta aggggaatta tttaaaaatg 2280
tttggggcaa gagatattga tggaacccta tttccaactt atctctatca aaaaatagat 2340
gagtccaggt taaaaccata tacacgttat cgagtaagag ggtttgtggg aagtagtaaa 2400
aatctaaaat tagtggtaac acgctatgag aaagaaattg atgccattat gaatgttcca 2460
aatgatttgg cacatatgca gcttaaccct tcatgtggag attatcgctg tgaatcatcg 2520
tcccagtttt tggtgaacca agtgcatcct acaccaacag ctggatatgc tcttgatatg 2580
tatgcatgcc cgtcaagttc agataaaaaa catattatgt gtcacgatcg tcatccattt 2640
gattttcata ttgacaccgg agaattaaat ccaaacacaa acctgggtat tgatgtcttg 2700
tttaaaattt ctaatccaaa tggatacgct acattaggga atctagaagt cattgaagaa 2760
ggaccactaa cagatgaagc attggtacat gtaaaacaaa aggaaaagaa atggcgtcag 2820
cacatggaga aaaaacgaat ggaaacacaa caagcctatg atccagcaaa acaagctgta 2880
gatgcattat ttacaaatga acaagagtta gactatcata ctactttaga tcatattcag 2940
aacgccgatc agctggtaca ggcgattccc tatgtacacc atgcttggtt accggatgct 3000
ccaggtatga actatgatgt atatcaaggg ttaaacgcac gtatcatgca ggcgtacaat 3060
ttatatgatg cacgaaatgt cataataaat ggtgacttta cacaaggact acaaggatgg 3120
cacgcaacag gaaaagcagc ggtacaacaa atagatggag cttcagtatt agttctatca 3180
aactggagtg ccgaggtatc tcagaatctg catgcccaag atcatcatgg atatatgtta 3240
cgtgtgattg ccaaaaaaga aggtcctgga aaagggtatg taatgatgat ggattttaat 3300
ggaaagcagg aaacacttac gttcacttct tgtgaagaag gatatataac aaaaacaata 3360
gaggtattcc cggaaagtga tcgaatacga attgaaatgg gagaaacaga gggtacgttt 3420
tatgtagata gcatcgagtt gctttgtatg caaggatatg ctagcgataa taacccgcac 3480
acgggtaata tgtatgagca aagttataat ggaaattata atcaaaatac tagcgatgtg 3540
tatcaccaag gatatataaa caactataac caaaattcta gtagtatgta taatcaaaat 3600
tatattaaca atgatgacct gcattccggt tgcacatgta accaagggca taactctggc 3660
tgtacatgta atcaaggata taaccgttag 3690
<210> 15
<211> 3690
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867_21.
<400> 15
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccggga ctaccaccta cgagtacgag gagaagcaga atctcgagaa ggctcagaag 1980
gctctgaacg ctctgttcac tgacgggacc aacggctacc tccagatgga cgccactgac 2040
tacgacatca accagacagc taacctgatt gagtgtgtga gtgacgaact gtacgctaag 2100
gagaagatcg tactcctgga cgaggtgaag tacgctaagc gcctgagcat tagccgtaac 2160
ctgctgctga acgacgatct ggagttcagc gacggctttg gcgagaacgg ctggaccacc 2220
agcgacaaca tctccatcca ggccgacaat ccactcttca aaggcaacta cctcaagatg 2280
ttcggagcca gggacatcga cggcaccctc tttccgacct acctctacca gaagatcgac 2340
gagtcccgcc tcaaacccta cacccgctac agggtgcgcg gcttcgtggg cagcagcaag 2400
aacctcaagc tcgtggtcac acggtatgag aaggagatcg acgccatcat gaacgtgccc 2460
aacgatctcg cccacatgca gctcaatcca tcctgcggcg actaccggtg cgagtccagc 2520
tcccagttcc tcgtgaacca ggtgcaccct actccgaccg ctggctatgc cctggacatg 2580
tacgcctgcc ctagttcctc cgacaagaag cacatcatgt gccacgaccg tcatccgttc 2640
gacttccaca tcgacaccgg cgaactgaac ccgaacacca acctgggcat cgacgtactg 2700
ttcaagattt ccaacccgaa cgggtacgcc accttgggca acctggaggt catcgaagaa 2760
ggcccgctga ccgacgaggc cctggtccac gtcaaacaga aggagaagaa gtggcggcag 2820
cacatggaga agaagcggat ggagactcaa caagcctacg acccggccaa gcaagctgtg 2880
gacgctctgt tcaccaacga gcaagagctt gactaccaca ctactcttga ccacatccag 2940
aatgctgacc agcttgtcca ggctattccg tacgtccacc acgcttggct accggacgct 3000
ccagggatga actacgatgt gtaccagggt ctgaacgcgc ggatcatgca agcgtacaac 3060
ctgtacgacg cgcgtaacgt catcatcaac ggtgacttca ctcagggtct tcaaggttgg 3120
cacgcgactg gcaaagcggc agtccagcag attgatggtg cgtctgttct tgtgttgagc 3180
aactggtctg cggaggtttc tcagaacctg cacgcacagg atcaccacgg ctacatgctg 3240
agggtgattg ctaagaagga gggccctggc aaaggctacg tcatgatgat ggacttcaac 3300
ggaaagcaag aaaccctgac cttcactagc tgtgaggagg gctacatcac taagaccatt 3360
gaggtctttc cggagtctga ccgcatccgg atcgagatgg gcgagaccga aggcacgttc 3420
tacgtggact ccatcgaact cctctgcatg caaggctacg cctccgacaa caacccacac 3480
acgggcaaca tgtacgagca gtcctacaac gggaactaca accagaacac ctccgatgtg 3540
taccatcagg gctacatcaa caactacaac cagaacagca gcagcatgta caaccagaac 3600
tacatcaaca acgatgactt gcactcgggt tgcacctgca accagggtca caacagtggg 3660
tgcacgtgca accagggata caaccgttga 3690
<210> 16
<211> 1229
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC867_21.
<400> 16
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Gly Thr Thr Thr Tyr Glu Tyr Glu Glu Lys Gln Asn Leu Glu
645 650 655
Lys Ala Gln Lys Ala Leu Asn Ala Leu Phe Thr Asp Gly Thr Asn Gly
660 665 670
Tyr Leu Gln Met Asp Ala Thr Asp Tyr Asp Ile Asn Gln Thr Ala Asn
675 680 685
Leu Ile Glu Cys Val Ser Asp Glu Leu Tyr Ala Lys Glu Lys Ile Val
690 695 700
Leu Leu Asp Glu Val Lys Tyr Ala Lys Arg Leu Ser Ile Ser Arg Asn
705 710 715 720
Leu Leu Leu Asn Asp Asp Leu Glu Phe Ser Asp Gly Phe Gly Glu Asn
725 730 735
Gly Trp Thr Thr Ser Asp Asn Ile Ser Ile Gln Ala Asp Asn Pro Leu
740 745 750
Phe Lys Gly Asn Tyr Leu Lys Met Phe Gly Ala Arg Asp Ile Asp Gly
755 760 765
Thr Leu Phe Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Arg Leu
770 775 780
Lys Pro Tyr Thr Arg Tyr Arg Val Arg Gly Phe Val Gly Ser Ser Lys
785 790 795 800
Asn Leu Lys Leu Val Val Thr Arg Tyr Glu Lys Glu Ile Asp Ala Ile
805 810 815
Met Asn Val Pro Asn Asp Leu Ala His Met Gln Leu Asn Pro Ser Cys
820 825 830
Gly Asp Tyr Arg Cys Glu Ser Ser Ser Gln Phe Leu Val Asn Gln Val
835 840 845
His Pro Thr Pro Thr Ala Gly Tyr Ala Leu Asp Met Tyr Ala Cys Pro
850 855 860
Ser Ser Ser Asp Lys Lys His Ile Met Cys His Asp Arg His Pro Phe
865 870 875 880
Asp Phe His Ile Asp Thr Gly Glu Leu Asn Pro Asn Thr Asn Leu Gly
885 890 895
Ile Asp Val Leu Phe Lys Ile Ser Asn Pro Asn Gly Tyr Ala Thr Leu
900 905 910
Gly Asn Leu Glu Val Ile Glu Glu Gly Pro Leu Thr Asp Glu Ala Leu
915 920 925
Val His Val Lys Gln Lys Glu Lys Lys Trp Arg Gln His Met Glu Lys
930 935 940
Lys Arg Met Glu Thr Gln Gln Ala Tyr Asp Pro Ala Lys Gln Ala Val
945 950 955 960
Asp Ala Leu Phe Thr Asn Glu Gln Glu Leu Asp Tyr His Thr Thr Leu
965 970 975
Asp His Ile Gln Asn Ala Asp Gln Leu Val Gln Ala Ile Pro Tyr Val
980 985 990
His His Ala Trp Leu Pro Asp Ala Pro Gly Met Asn Tyr Asp Val Tyr
995 1000 1005
Gln Gly Leu Asn Ala Arg Ile Met Gln Ala Tyr Asn Leu Tyr Asp
1010 1015 1020
Ala Arg Asn Val Ile Ile Asn Gly Asp Phe Thr Gln Gly Leu Gln
1025 1030 1035
Gly Trp His Ala Thr Gly Lys Ala Ala Val Gln Gln Ile Asp Gly
1040 1045 1050
Ala Ser Val Leu Val Leu Ser Asn Trp Ser Ala Glu Val Ser Gln
1055 1060 1065
Asn Leu His Ala Gln Asp His His Gly Tyr Met Leu Arg Val Ile
1070 1075 1080
Ala Lys Lys Glu Gly Pro Gly Lys Gly Tyr Val Met Met Met Asp
1085 1090 1095
Phe Asn Gly Lys Gln Glu Thr Leu Thr Phe Thr Ser Cys Glu Glu
1100 1105 1110
Gly Tyr Ile Thr Lys Thr Ile Glu Val Phe Pro Glu Ser Asp Arg
1115 1120 1125
Ile Arg Ile Glu Met Gly Glu Thr Glu Gly Thr Phe Tyr Val Asp
1130 1135 1140
Ser Ile Glu Leu Leu Cys Met Gln Gly Tyr Ala Ser Asp Asn Asn
1145 1150 1155
Pro His Thr Gly Asn Met Tyr Glu Gln Ser Tyr Asn Gly Asn Tyr
1160 1165 1170
Asn Gln Asn Thr Ser Asp Val Tyr His Gln Gly Tyr Ile Asn Asn
1175 1180 1185
Tyr Asn Gln Asn Ser Ser Ser Met Tyr Asn Gln Asn Tyr Ile Asn
1190 1195 1200
Asn Asp Asp Leu His Ser Gly Cys Thr Cys Asn Gln Gly His Asn
1205 1210 1215
Ser Gly Cys Thr Cys Asn Gln Gly Tyr Asn Arg
1220 1225
<210> 17
<211> 3432
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC867_22.
<400> 17
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta cacaaatacc attggtaaag gcgcataccc tccaatcggg taccactgta 1560
gtaaaagggc cagggtttac aggaggggat atcctccgtc gaacaagtgg aggaccattt 1620
gcttttagta atgttaatct agattttaac ttgtcacaaa ggtatcgtgc tagaattcgt 1680
tatgcctcta ctactaacct aagaatttac gtaacggttg caggtgaacg aatttttgct 1740
ggtcaatttg acaaaactat ggatgctggt gccccattaa cattccaatc ttttagttac 1800
gcaactatta atacagcttt tacattccca gaaagatcga gcagcttgac tgtaggtgcc 1860
gatacgttta gttcaggtaa tgaagtttat gtagatagat ttgaattaat cccagttact 1920
gcaaccaatc cgacgcgaga ggcggaagag gatctagaag cagcgaagaa agcggtggcg 1980
agcttgttta cacgtacaag ggacggatta caagtaaatg tgacagatta tcaagtcgat 2040
caagcggcaa atttagtgtc atgcttatca gatgaacaat atgggcatga caaaaagatg 2100
ttattggaag cggtaagagc ggcaaaacgc ctcagccgag aacgcaactt acttcaggat 2160
ccagatttta atacaatcaa tagtacagaa gaaaatggat ggaaagcaag taacggcgtt 2220
actattagcg agggcggtcc attctataaa ggccgtgcgc ttcagctagc aagcgcaaga 2280
gaaaattacc caacatacat ttatcaaaaa gtaaatgcat cagagttaaa gccgtataca 2340
cgttatagac tggatgggtt cgtgaagagt agtcaagatt tagaaattga tctcattcac 2400
catcataaag tccatctcgt gaaaaatgta ccagataatt tagtatccga tacttactcg 2460
gatggttctt gcagtggaat gaatcgatgt gaggaacaac agatggtaaa tgcgcaactg 2520
gaaacagaac atcatcatcc gatggattgc tgtgaagcgg ctcaaacaca tgagttttct 2580
tcctatatta atacaggcga tctaaattca agtgtagatc aaggcatttg ggttgtattg 2640
aaagttcgaa caaccgatgg ttatgcgacg ctaggaaatc ttgaattggt agaggtcgga 2700
ccgttatcgg gtgaatctct agaacgtgaa caaagggata atgcgaaatg gagtgcagag 2760
ctaggaagaa agcgtgcaga aacagatcgc gtgtatcaag atgccaaaca atccatcaat 2820
catttatttg tggattatca agatcaacaa ttaaatccag aaatagggat ggcagatatt 2880
attgacgctc aaaatcttgt cgcatcaatt tcagatgtgt atagcgatgc agtactgcaa 2940
atccctggaa ttaactatga gatttacaca gagctatcca atcgcttaca acaagcatcg 3000
tatctgtata cgtctcgaaa tgcggtgcaa aatggggact ttaacagcgg tctagatagt 3060
tggaatgcaa cagggggggc tacggtacaa caggatggca atacgcattt cttagttctt 3120
tctcattggg atgcacaagt ttctcaacaa tttagagtgc agccgaattg taaatatgta 3180
ttacgtgtaa cagcagagaa agtaggcggc ggagacggat acgtgacaat ccgggatggt 3240
gctcatcata cagaaaagct tacatttaat gcatgtgatt atgatataaa tggcacgtac 3300
gtgactgata atacgtatct aacaaaagaa gtggtattct attcacatac agaacacatg 3360
tgggtagagg taagtgaaac agaaggtgca tttcatatag atagtattga attcgttgaa 3420
acagaaaagt ag 3432
<210> 18
<211> 3432
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867_22.
<400> 18
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccaacc cgacgcggga agctgaggaa gacttggaag ccgccaagaa agcggtcgcc 1980
agcctgttta ctcggacgcg ggacgggctc caagtgaatg tgacggacta tcaagtggat 2040
caggccgcta acctcgtgtc atgcctgagc gacgagcagt acggtcacga caagaaaatg 2100
ctgctggagg ccgtccgggc cgccaagcgg ctgtccaggg agcgtaacct gctacaagat 2160
cccgacttta acacgatcaa cagcacagag gagaatggct ggaaggccag caacggagtt 2220
acgataagcg agggcggtcc gttctacaag ggtcgtgccc tccagctcgc ctctgcaagg 2280
gagaactatc caacctacat ctatcagaag gtgaacgcat ccgagcttaa gccctacaca 2340
cgctaccgcc tggacgggtt cgttaagtcc agtcaagacc tagagataga cctcatccac 2400
caccacaaag tgcatctggt caagaacgtt cccgataatc tcgtgagcga tacctactca 2460
gacggctcat gctctggcat gaacagatgt gaggagcaac agatggttaa tgctcaactc 2520
gaaaccgagc atcatcatcc tatggattgc tgcgaggccg cgcagaccca tgagttcagc 2580
tcttacatca acaccggaga cctcaacagt agcgtggatc agggaatttg ggtggtgctt 2640
aaagtgcgta caaccgacgg ctacgccacc ctcggcaacc ttgagcttgt cgaggtcgga 2700
ccacttagcg gcgagtccct ggaacgtgag cagcgggaca acgccaaatg gagcgcagag 2760
ctagggcgca aacgcgcgga gacggaccgg gtttatcagg acgcgaagca gtccatcaat 2820
cacctcttcg tggattatca ggaccagcag cttaatccag agatcggcat ggccgacatc 2880
atcgacgccc agaacctagt agcgtcgatt tccgatgtct attccgacgc cgtgcttcaa 2940
atacctggca tcaactacga gatctacaca gagttgtcca acaggctcca gcaagcgtca 3000
tacctctaca ccagccgcaa cgccgtccag aatggcgact tcaattccgg actagactcc 3060
tggaacgcca cgggcggagc tacggtgcaa caagacggca acacccactt cctcgtactt 3120
agccactggg acgctcaagt gagtcagcaa ttccgggttc agccgaactg caagtacgtc 3180
ctgcgcgtaa cggccgagaa ggttggaggc ggagacggct acgttaccat ccgcgacggc 3240
gctcaccaca ccgagaaact gacgttcaac gcttgtgact acgacatcaa cggcacttac 3300
gtgacggaca acacctacct gacgaaggag gtggtgttct attctcacac cgagcacatg 3360
tgggttgagg tcagcgagac cgagggagcc ttccacattg acagcatcga gttcgtggag 3420
actgagaagt ga 3432
<210> 19
<211> 1143
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC867_22.
<400> 19
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Asn Pro Thr Arg Glu Ala Glu Glu Asp Leu Glu Ala Ala Lys
645 650 655
Lys Ala Val Ala Ser Leu Phe Thr Arg Thr Arg Asp Gly Leu Gln Val
660 665 670
Asn Val Thr Asp Tyr Gln Val Asp Gln Ala Ala Asn Leu Val Ser Cys
675 680 685
Leu Ser Asp Glu Gln Tyr Gly His Asp Lys Lys Met Leu Leu Glu Ala
690 695 700
Val Arg Ala Ala Lys Arg Leu Ser Arg Glu Arg Asn Leu Leu Gln Asp
705 710 715 720
Pro Asp Phe Asn Thr Ile Asn Ser Thr Glu Glu Asn Gly Trp Lys Ala
725 730 735
Ser Asn Gly Val Thr Ile Ser Glu Gly Gly Pro Phe Tyr Lys Gly Arg
740 745 750
Ala Leu Gln Leu Ala Ser Ala Arg Glu Asn Tyr Pro Thr Tyr Ile Tyr
755 760 765
Gln Lys Val Asn Ala Ser Glu Leu Lys Pro Tyr Thr Arg Tyr Arg Leu
770 775 780
Asp Gly Phe Val Lys Ser Ser Gln Asp Leu Glu Ile Asp Leu Ile His
785 790 795 800
His His Lys Val His Leu Val Lys Asn Val Pro Asp Asn Leu Val Ser
805 810 815
Asp Thr Tyr Ser Asp Gly Ser Cys Ser Gly Met Asn Arg Cys Glu Glu
820 825 830
Gln Gln Met Val Asn Ala Gln Leu Glu Thr Glu His His His Pro Met
835 840 845
Asp Cys Cys Glu Ala Ala Gln Thr His Glu Phe Ser Ser Tyr Ile Asn
850 855 860
Thr Gly Asp Leu Asn Ser Ser Val Asp Gln Gly Ile Trp Val Val Leu
865 870 875 880
Lys Val Arg Thr Thr Asp Gly Tyr Ala Thr Leu Gly Asn Leu Glu Leu
885 890 895
Val Glu Val Gly Pro Leu Ser Gly Glu Ser Leu Glu Arg Glu Gln Arg
900 905 910
Asp Asn Ala Lys Trp Ser Ala Glu Leu Gly Arg Lys Arg Ala Glu Thr
915 920 925
Asp Arg Val Tyr Gln Asp Ala Lys Gln Ser Ile Asn His Leu Phe Val
930 935 940
Asp Tyr Gln Asp Gln Gln Leu Asn Pro Glu Ile Gly Met Ala Asp Ile
945 950 955 960
Ile Asp Ala Gln Asn Leu Val Ala Ser Ile Ser Asp Val Tyr Ser Asp
965 970 975
Ala Val Leu Gln Ile Pro Gly Ile Asn Tyr Glu Ile Tyr Thr Glu Leu
980 985 990
Ser Asn Arg Leu Gln Gln Ala Ser Tyr Leu Tyr Thr Ser Arg Asn Ala
995 1000 1005
Val Gln Asn Gly Asp Phe Asn Ser Gly Leu Asp Ser Trp Asn Ala
1010 1015 1020
Thr Gly Gly Ala Thr Val Gln Gln Asp Gly Asn Thr His Phe Leu
1025 1030 1035
Val Leu Ser His Trp Asp Ala Gln Val Ser Gln Gln Phe Arg Val
1040 1045 1050
Gln Pro Asn Cys Lys Tyr Val Leu Arg Val Thr Ala Glu Lys Val
1055 1060 1065
Gly Gly Gly Asp Gly Tyr Val Thr Ile Arg Asp Gly Ala His His
1070 1075 1080
Thr Glu Lys Leu Thr Phe Asn Ala Cys Asp Tyr Asp Ile Asn Gly
1085 1090 1095
Thr Tyr Val Thr Asp Asn Thr Tyr Leu Thr Lys Glu Val Val Phe
1100 1105 1110
Tyr Ser His Thr Glu His Met Trp Val Glu Val Ser Glu Thr Glu
1115 1120 1125
Gly Ala Phe His Ile Asp Ser Ile Glu Phe Val Glu Thr Glu Lys
1130 1135 1140
<210> 20
<211> 3696
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867_23.
<400> 20
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccacgg cgaccttcga ggcggagtat gacttggagc gggctcagga ggccgtcaac 1980
gcgctgttca caaacaccaa tcctcgccgc ctcaagacgg gtgtgactga ttaccacatt 2040
gacgaggtct ccaacttggt cgcgtgtctg tccgatgagt tctgcctgga cgagaagcgg 2100
gaactgctgg agaaggtcaa gtacgccaag cgcctctccg acgaaaggaa cctcctccaa 2160
gatcccaact ttacttccat taacaagcag ccggacttca tctccaccaa cgagcagtcc 2220
aacttcacct caatccacga gcagtcggag cacgggtggt ggggcagcga gaacatcacc 2280
atccaagagg gcaacgacgt cttcaaggag aactacgtga tcctgcccgg caccttcaac 2340
gagtgttacc cgacctatct ctaccagaag attggcgaag cggaactcaa ggcttacacc 2400
cgttaccaac tgagtggcta cattgaggac tcacaagacc tggaaatcta cctgatccgc 2460
tacaacgcca agcacgagac cctcgacgtg cctggcacgg agtccgtctg gcccttgagc 2520
gtggagtctc ctatcggtcg ttgcggcgag cccaatcgct gcgctccgca ctttgagtgg 2580
aatcctgatt tggattgctc ctgccgagac ggtgagaaat gcgcccacca ctcgcaccac 2640
ttcagcctag acatcgacgt gggctgcatc gacctgcacg agaacttggg cgtctgggtc 2700
gtgttcaaga tcaagacaca ggagggccat gctcggcttg ggaacctgga gttcatcgag 2760
gagaagccac tgctgggtga agccttgtca cgggtgaaac gcgccgagaa gaagtggcgg 2820
gacaaacggg agaagctcca gttggagaca aagcgtgtgt acacagaggc caaggaggcc 2880
gtggatgcct tgttcgtgga cagtcagtac gacaggctgc aagcggacac caacatcggg 2940
atgatccacg cggctgataa gcttgttcac agaatccgcg aggcgtacct gtcagagctt 3000
agcgtgatcc caggcgtcaa cgccgaaatc ttcgaggaac tggagggccg cattatcacg 3060
gcaatctcac tttatgacgc gaggaatgtg gtcaagaacg gtgacttcaa caacggcttg 3120
gcgtgttgga acgttaaagg gcacgtggat gtacaacagt cacaccacag aagtgtcttg 3180
gtcatcccgg agtgggaggc ggaagtgagc caggccgtcc gggtctgccc tgggcgcggt 3240
tacatcctcc gcgtgacagc gtacaaggag ggctacggtg agggctgcgt gacgatccac 3300
gagattgaga acaacacgga cgagcttaag ttcaagaact gcgaggagga ggaagtgtac 3360
ccgacagaca ccggcacctg caacgactac accgcccacc aagggaccgc cgcctgcaac 3420
agccgcaacg cgggctatga agatgcgtac gaggttgata ccaccgcctc agtgaactac 3480
aaaccgactt atgaggagga gacatacacg gacgtcaggc gcgacaacca ttgtgagtac 3540
gaccgtggct acgtgaacta tccgccggtg ccagcgggct acatgacgaa ggagctagaa 3600
tacttccctg agacggacaa ggtgtggatt gaaatcggcg agaccgaggg caagtttatc 3660
gtggattctg tcgagctgct gctaatggag gagtag 3696
<210> 21
<211> 1231
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC867_23.
<400> 21
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Thr Ala Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg Ala Gln
645 650 655
Glu Ala Val Asn Ala Leu Phe Thr Asn Thr Asn Pro Arg Arg Leu Lys
660 665 670
Thr Gly Val Thr Asp Tyr His Ile Asp Glu Val Ser Asn Leu Val Ala
675 680 685
Cys Leu Ser Asp Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Leu Glu
690 695 700
Lys Val Lys Tyr Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln
705 710 715 720
Asp Pro Asn Phe Thr Ser Ile Asn Lys Gln Pro Asp Phe Ile Ser Thr
725 730 735
Asn Glu Gln Ser Asn Phe Thr Ser Ile His Glu Gln Ser Glu His Gly
740 745 750
Trp Trp Gly Ser Glu Asn Ile Thr Ile Gln Glu Gly Asn Asp Val Phe
755 760 765
Lys Glu Asn Tyr Val Ile Leu Pro Gly Thr Phe Asn Glu Cys Tyr Pro
770 775 780
Thr Tyr Leu Tyr Gln Lys Ile Gly Glu Ala Glu Leu Lys Ala Tyr Thr
785 790 795 800
Arg Tyr Gln Leu Ser Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile
805 810 815
Tyr Leu Ile Arg Tyr Asn Ala Lys His Glu Thr Leu Asp Val Pro Gly
820 825 830
Thr Glu Ser Val Trp Pro Leu Ser Val Glu Ser Pro Ile Gly Arg Cys
835 840 845
Gly Glu Pro Asn Arg Cys Ala Pro His Phe Glu Trp Asn Pro Asp Leu
850 855 860
Asp Cys Ser Cys Arg Asp Gly Glu Lys Cys Ala His His Ser His His
865 870 875 880
Phe Ser Leu Asp Ile Asp Val Gly Cys Ile Asp Leu His Glu Asn Leu
885 890 895
Gly Val Trp Val Val Phe Lys Ile Lys Thr Gln Glu Gly His Ala Arg
900 905 910
Leu Gly Asn Leu Glu Phe Ile Glu Glu Lys Pro Leu Leu Gly Glu Ala
915 920 925
Leu Ser Arg Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu
930 935 940
Lys Leu Gln Leu Glu Thr Lys Arg Val Tyr Thr Glu Ala Lys Glu Ala
945 950 955 960
Val Asp Ala Leu Phe Val Asp Ser Gln Tyr Asp Arg Leu Gln Ala Asp
965 970 975
Thr Asn Ile Gly Met Ile His Ala Ala Asp Lys Leu Val His Arg Ile
980 985 990
Arg Glu Ala Tyr Leu Ser Glu Leu Ser Val Ile Pro Gly Val Asn Ala
995 1000 1005
Glu Ile Phe Glu Glu Leu Glu Gly Arg Ile Ile Thr Ala Ile Ser
1010 1015 1020
Leu Tyr Asp Ala Arg Asn Val Val Lys Asn Gly Asp Phe Asn Asn
1025 1030 1035
Gly Leu Ala Cys Trp Asn Val Lys Gly His Val Asp Val Gln Gln
1040 1045 1050
Ser His His Arg Ser Val Leu Val Ile Pro Glu Trp Glu Ala Glu
1055 1060 1065
Val Ser Gln Ala Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu
1070 1075 1080
Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr
1085 1090 1095
Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Lys Asn
1100 1105 1110
Cys Glu Glu Glu Glu Val Tyr Pro Thr Asp Thr Gly Thr Cys Asn
1115 1120 1125
Asp Tyr Thr Ala His Gln Gly Thr Ala Ala Cys Asn Ser Arg Asn
1130 1135 1140
Ala Gly Tyr Glu Asp Ala Tyr Glu Val Asp Thr Thr Ala Ser Val
1145 1150 1155
Asn Tyr Lys Pro Thr Tyr Glu Glu Glu Thr Tyr Thr Asp Val Arg
1160 1165 1170
Arg Asp Asn His Cys Glu Tyr Asp Arg Gly Tyr Val Asn Tyr Pro
1175 1180 1185
Pro Val Pro Ala Gly Tyr Met Thr Lys Glu Leu Glu Tyr Phe Pro
1190 1195 1200
Glu Thr Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Lys
1205 1210 1215
Phe Ile Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1220 1225 1230
<210> 22
<211> 3666
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867_24.
<400> 22
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccaccg cgacgtttga agctgaatcc gacctcgagc gtgcgcgcaa ggcggtgaac 1980
gctctgttca cgagcaccaa ccctcgtggc ttgaagacgg atgtgacgga ctaccacatc 2040
gaccaagtct cgaacctcgt ggagtgcctg agcgacgagt tctgtcttga caagaagcgc 2100
gagctgctgg aggaggtgaa gtacgccaag cgcctctccg atgagcgcaa cctgctccaa 2160
gatcctacct tcacgtcgat ttccggccaa accgaccgtg gatggatcgg ctcgactggc 2220
atctccatcc agggcggcga cgacatcttc aaggagaact atgttcggct gccgggcacg 2280
gtggacgagt gttacccgac gtacctctac cagaagatag acgagagtca actcaagtcc 2340
tacacgcggt atcagttacg tggctacatt gaagactccc aggacttgga aatctatctc 2400
atacggtaca acgccaagca cgagacctta agcgtgccgg gaacggagtc gccctggcca 2460
agctctggcg tgtacccttc cggtaggtgc ggcgagccca accgctgtgc acctcgaatc 2520
gaatggaacc cggaccttga ctgctcttgc cggtacggcg agaagtgcgt ccatcattct 2580
caccacttca gcttggacat tgacgtcggc tgcaccgacc tcaatgaaga cctcggagtg 2640
tgggtcatct tcaagatcaa gacacaggac gggcacgcga aactaggaaa cctggagttc 2700
atcgaggaga agccactcct cggcaaggca ctttccaggg tcaagcgggc cgagaagaaa 2760
tggagggaca agtacgagaa actccagctc gaaacaaagc gggtgtacac ggaggcaaag 2820
gaatccgtgg acgccctgtt cgtggactct cagtacgaca agctccaggc gaacacaaac 2880
attggcatca tccacggtgc ggacaagcaa gtgcacagga tacgggagcc ttacctctcg 2940
gagctgccgg tgattccctc gatcaacgcg gcgatcttcg aggaactgga gggccacatc 3000
ttcaaggcgt attctctgta cgacgcgcgt aacgtcatca agaacggcga cttcaacaat 3060
gggctgtcct gctggaacgt taaaggccac gtcgatgtcc agcagaacca ccataggtca 3120
gtcctggtgc tgagcgagtg ggaggcggag gtgtcccaga aggtgcgcgt gtgcccggat 3180
cgcggctaca tcttgagggt gacagcctac aaggagggct acggcgaggg ctgtgtcacg 3240
atccatgagt tcgaggacaa cacggatgtc ctgaaattcc gtaacttcgt cgaggaggag 3300
gtctatccca acaacaccgt gacctgcaac gactacacga ccaatcagtc ggctgagggc 3360
agtaccgatg cctgcaacag ctacaaccgt ggttacgaag atggatacga gaaccgctac 3420
gagcccaatc cttcggctcc cgtgaattac actcccacgt acgaggaggg catgtacact 3480
gacactcagg gctacaacca ttgcgtcagc gaccgtggct accgcaacca cacgccgctc 3540
ccagcgggct acgtgacgct ggagctggaa tactttcccg agacagaaca agtgtggata 3600
gagatcggcg agaccgaggg cacattcatc gtgggctctg tggaattgct cctcatggag 3660
gagtaa 3666
<210> 23
<211> 1221
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC867_24.
<400> 23
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Thr Ala Thr Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala Arg
645 650 655
Lys Ala Val Asn Ala Leu Phe Thr Ser Thr Asn Pro Arg Gly Leu Lys
660 665 670
Thr Asp Val Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Glu
675 680 685
Cys Leu Ser Asp Glu Phe Cys Leu Asp Lys Lys Arg Glu Leu Leu Glu
690 695 700
Glu Val Lys Tyr Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln
705 710 715 720
Asp Pro Thr Phe Thr Ser Ile Ser Gly Gln Thr Asp Arg Gly Trp Ile
725 730 735
Gly Ser Thr Gly Ile Ser Ile Gln Gly Gly Asp Asp Ile Phe Lys Glu
740 745 750
Asn Tyr Val Arg Leu Pro Gly Thr Val Asp Glu Cys Tyr Pro Thr Tyr
755 760 765
Leu Tyr Gln Lys Ile Asp Glu Ser Gln Leu Lys Ser Tyr Thr Arg Tyr
770 775 780
Gln Leu Arg Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu
785 790 795 800
Ile Arg Tyr Asn Ala Lys His Glu Thr Leu Ser Val Pro Gly Thr Glu
805 810 815
Ser Pro Trp Pro Ser Ser Gly Val Tyr Pro Ser Gly Arg Cys Gly Glu
820 825 830
Pro Asn Arg Cys Ala Pro Arg Ile Glu Trp Asn Pro Asp Leu Asp Cys
835 840 845
Ser Cys Arg Tyr Gly Glu Lys Cys Val His His Ser His His Phe Ser
850 855 860
Leu Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val
865 870 875 880
Trp Val Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala Lys Leu Gly
885 890 895
Asn Leu Glu Phe Ile Glu Glu Lys Pro Leu Leu Gly Lys Ala Leu Ser
900 905 910
Arg Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Tyr Glu Lys Leu
915 920 925
Gln Leu Glu Thr Lys Arg Val Tyr Thr Glu Ala Lys Glu Ser Val Asp
930 935 940
Ala Leu Phe Val Asp Ser Gln Tyr Asp Lys Leu Gln Ala Asn Thr Asn
945 950 955 960
Ile Gly Ile Ile His Gly Ala Asp Lys Gln Val His Arg Ile Arg Glu
965 970 975
Pro Tyr Leu Ser Glu Leu Pro Val Ile Pro Ser Ile Asn Ala Ala Ile
980 985 990
Phe Glu Glu Leu Glu Gly His Ile Phe Lys Ala Tyr Ser Leu Tyr Asp
995 1000 1005
Ala Arg Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser
1010 1015 1020
Cys Trp Asn Val Lys Gly His Val Asp Val Gln Gln Asn His His
1025 1030 1035
Arg Ser Val Leu Val Leu Ser Glu Trp Glu Ala Glu Val Ser Gln
1040 1045 1050
Lys Val Arg Val Cys Pro Asp Arg Gly Tyr Ile Leu Arg Val Thr
1055 1060 1065
Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu
1070 1075 1080
Phe Glu Asp Asn Thr Asp Val Leu Lys Phe Arg Asn Phe Val Glu
1085 1090 1095
Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp Tyr Thr
1100 1105 1110
Thr Asn Gln Ser Ala Glu Gly Ser Thr Asp Ala Cys Asn Ser Tyr
1115 1120 1125
Asn Arg Gly Tyr Glu Asp Gly Tyr Glu Asn Arg Tyr Glu Pro Asn
1130 1135 1140
Pro Ser Ala Pro Val Asn Tyr Thr Pro Thr Tyr Glu Glu Gly Met
1145 1150 1155
Tyr Thr Asp Thr Gln Gly Tyr Asn His Cys Val Ser Asp Arg Gly
1160 1165 1170
Tyr Arg Asn His Thr Pro Leu Pro Ala Gly Tyr Val Thr Leu Glu
1175 1180 1185
Leu Glu Tyr Phe Pro Glu Thr Glu Gln Val Trp Ile Glu Ile Gly
1190 1195 1200
Glu Thr Glu Gly Thr Phe Ile Val Gly Ser Val Glu Leu Leu Leu
1205 1210 1215
Met Glu Glu
1220
<210> 24
<211> 3651
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC867_25.
<400> 24
atgaccagca accgaaagaa cgagaacgag atcatcaacg ccctgtccat accggccgtg 60
tcaaaccact ccgcccagat gaacctctcc accgacgcga ggatcgagga ctccctctgc 120
atcgccgagg gcaacaacat cgacccgttc gtgtctgcaa gcacggtcca gaccggcatc 180
aacatcgcgg gccgcatcct gggcgtgctc ggcgtgccct tcgcgggtca aatcgcctct 240
ttctactcat tcctcgtggg cgagctgtgg ccgcgcggac gtgacccgtg ggaaatcttc 300
ctggagcacg ttgagcagct catccggcag caagtgaccg agaacaccag ggacaccgca 360
ctggcacggc tccagggcct tggcaacagc ttccgcgcct accagcagtc gctggaggac 420
tggctggaga accgagacga cgccagaacc cgctcagttc tgtacacaca gtacatcgcc 480
ctagagctgg acttcctcaa cgctatgccg ctcttcgcca tccgtaacca ggaagtaccg 540
cttctgatgg tgtacgcaca agcagcgaac ctccatctgc tcctgctgcg agacgcatct 600
ctgttcggca gtgagttcgg gctgacgagc caggagatcc agcgctacta cgagcgccaa 660
gtggagaaga ctcgtgagta cagcgactac tgcgcgcgct ggtacaacac gggcttgaac 720
aaccttcgcg ggacaaacgc cgaatcctgg cttcgctaca accagttccg ccgcgacctc 780
acgctgggtg tgctggacct ggtcgcgctc ttcccgtcct acgacacacg ggtgtaccca 840
atgaacacga gcgcacagct cacccgtgag atctacacag atcccatcgg ccgcaccaac 900
gctcccagtg gcttcgcaag cacgaattgg ttcaacaata acgctccttc tttctctgcc 960
atcgaggccg ctgtcatcag accgccgcac ttactcgatt tcccggagca gctcactatc 1020
ttctctgtgt tgtcccggtg gtcgaacacg cagtacatga actactgggt gggccacagg 1080
ctagagagcc ggaccatccg tggcagtctc tcaacctcga cccacggcaa cacgaacacg 1140
agcatcaacc ctgtcactct ccagtttaca tctagggacg tttacaggac agagtcgttc 1200
gctggcatta acattctgtt gaccactccg gtgaacggcg tcccttgggc ccgcttcaac 1260
tggaggaatc ctctgaactc actgcgcggc agccttctct acactatcgg ctacaccggc 1320
gttgggacgc aactcttcga ctcggagacc gagctgccgc ccgagaccac cgagcggcct 1380
aactacgaga gttattcaca caggctctcc aacatccgct tgatttctgg gaacaccttg 1440
cgggctccgg tgtactcctg gacgcaccgc agcgccgaca gaactaatac catcagctcc 1500
gactcgatca cccagatccc gctggtgaag gctcacacgc ttcagtcggg caccacagtc 1560
gtcaagggcc ctggcttcac cggcggcgac atcctgcgtc gcacatctgg cggacccttc 1620
gccttcagca acgtgaactt ggacttcaat ttgtcacagc ggtatcgtgc cagaatccgg 1680
tacgccagca ctacgaacct gcgaatctat gttactgtgg cgggcgagcg gatcttcgcc 1740
gggcaattcg acaagacgat ggacgcggga gcacctctga cattccagtc attctcttac 1800
gccacgatca acacggcatt cacgtttccg gagcgttcca gtagcctgac cgtgggcgct 1860
gataccttca gtagcgggaa cgaggtgtac gttgaccgtt tcgagctgat cccggtcacc 1920
gccaccgatg ctacctttga agcagagtcc gacttggaac gtgcacagaa ggcagtgaac 1980
gcactcttca cctcaagcaa ccagatcgga ttgaagacag atgtgacaga ttaccacatc 2040
gaccaagtga gcaacttggt ggattgcttg tcagatgagt tctgcttgga tgagaagcgt 2100
gaactctccg agaaggtgaa gcacgcaaag cgtctctcag atgaacgtaa tctccttcaa 2160
gaccctaact ttcgtggtat caatcgtcag ccagatcgtg gatggcgtgg atcaacagac 2220
atcaccatcc agggaggcga tgatgtgttc aaggagaact acgtgaccct cccaggaacc 2280
gtggatgaat gctacccaac ctacctctac cagaagatcg acgagtcaaa gctcaaggct 2340
tacacccgtt atgaactccg tggctacatc gaagatagcc aggatctcga aatctatctc 2400
atccgttaca atgctaagca cgaaatcgtg aatgtgccag gaaccggctc actctggcca 2460
ctctcagcac agtcaccaat cggcaagtgc ggcgaaccca atcgctgcgc tcctcatctc 2520
gaatggaatc ccgatctcga ctgctcctgc cgagacggcg agaagtgtgc acatcactca 2580
caccacttca ccctcgacat cgacgtgggc tgcaccgacc tcaatgaaga cctgggcgtg 2640
tgggtgatct tcaagatcaa gacccaggac ggccacgcac gactgggcaa tctggagttt 2700
ctggaggaga agccactgct tggcgaggca ctggcacgag tgaaacgagc cgagaagaaa 2760
tggcgagaca aacgtgagaa gctgcaactg gagaccaaca tcgtgtacaa agaggccaaa 2820
gagtcagttg acgccctgtt tgtcaatagc cagtatgacc gactgcaagt tgacaccaac 2880
atcgccatga tccacgctgc ggacaagcgc gtccaccgca tccgcgaggc ttatctgccc 2940
gagctgagcg tcattcccgg cgtcaatgcc gcgatcttcg aggagttaga gggccgcatc 3000
ttcaccgcct acagcctcta tgacgcccgc aatgtcatta agaatggcga cttcaacaat 3060
ggcttactat gctggaatgt caaagggcac gttgacgtcg aggagcagaa caatcaccgc 3120
agcgtcttag tcatacccga gtgggaggcc gaagtcagcc aggaagtccg cgtctgtcca 3180
gggcgcgggt acatcctgcg ggtcaccgcc tacaaagagg gatacggcga gggttgtgtc 3240
accatacacg agatagagga caataccgac gaactcaagt tcagcaattg tgtcgaggag 3300
gaagtctatc ccaacaatac cgtaacctgc aacaactaca ccggaaccca ggaggagtat 3360
gaagggacgt acacctcgcg gaaccagggc tatgacgaag cctatgggaa caacccgtcg 3420
gtgcctgctg actatgcgtc ggtctatgag gagaaatcgt acacggacgg gcggcgggag 3480
aatccgtgtg agtcgaatcg cgggtatggt gactacacgc cgctaccggc gggctatgta 3540
acgaaagacc tggaatactt cccggagacg gacaaagtat ggatagagat aggcgagacg 3600
gagggaacgt tcatcgtgga ctcggtagag ctgctgctca tggaggagtg a 3651
<210> 25
<211> 1216
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC867_25.
<400> 25
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Thr Gln Ile Pro Leu Val Lys Ala His
500 505 510
Thr Leu Gln Ser Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Thr Ser Gly Gly Pro Phe Ala Phe Ser Asn
530 535 540
Val Asn Leu Asp Phe Asn Leu Ser Gln Arg Tyr Arg Ala Arg Ile Arg
545 550 555 560
Tyr Ala Ser Thr Thr Asn Leu Arg Ile Tyr Val Thr Val Ala Gly Glu
565 570 575
Arg Ile Phe Ala Gly Gln Phe Asp Lys Thr Met Asp Ala Gly Ala Pro
580 585 590
Leu Thr Phe Gln Ser Phe Ser Tyr Ala Thr Ile Asn Thr Ala Phe Thr
595 600 605
Phe Pro Glu Arg Ser Ser Ser Leu Thr Val Gly Ala Asp Thr Phe Ser
610 615 620
Ser Gly Asn Glu Val Tyr Val Asp Arg Phe Glu Leu Ile Pro Val Thr
625 630 635 640
Ala Thr Asp Ala Thr Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala Gln
645 650 655
Lys Ala Val Asn Ala Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu Lys
660 665 670
Thr Asp Val Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Asp
675 680 685
Cys Leu Ser Asp Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Ser Glu
690 695 700
Lys Val Lys His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln
705 710 715 720
Asp Pro Asn Phe Arg Gly Ile Asn Arg Gln Pro Asp Arg Gly Trp Arg
725 730 735
Gly Ser Thr Asp Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu
740 745 750
Asn Tyr Val Thr Leu Pro Gly Thr Val Asp Glu Cys Tyr Pro Thr Tyr
755 760 765
Leu Tyr Gln Lys Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg Tyr
770 775 780
Glu Leu Arg Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu
785 790 795 800
Ile Arg Tyr Asn Ala Lys His Glu Ile Val Asn Val Pro Gly Thr Gly
805 810 815
Ser Leu Trp Pro Leu Ser Ala Gln Ser Pro Ile Gly Lys Cys Gly Glu
820 825 830
Pro Asn Arg Cys Ala Pro His Leu Glu Trp Asn Pro Asp Leu Asp Cys
835 840 845
Ser Cys Arg Asp Gly Glu Lys Cys Ala His His Ser His His Phe Thr
850 855 860
Leu Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val
865 870 875 880
Trp Val Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly
885 890 895
Asn Leu Glu Phe Leu Glu Glu Lys Pro Leu Leu Gly Glu Ala Leu Ala
900 905 910
Arg Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu
915 920 925
Gln Leu Glu Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp
930 935 940
Ala Leu Phe Val Asn Ser Gln Tyr Asp Arg Leu Gln Val Asp Thr Asn
945 950 955 960
Ile Ala Met Ile His Ala Ala Asp Lys Arg Val His Arg Ile Arg Glu
965 970 975
Ala Tyr Leu Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile
980 985 990
Phe Glu Glu Leu Glu Gly Arg Ile Phe Thr Ala Tyr Ser Leu Tyr Asp
995 1000 1005
Ala Arg Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Leu
1010 1015 1020
Cys Trp Asn Val Lys Gly His Val Asp Val Glu Glu Gln Asn Asn
1025 1030 1035
His Arg Ser Val Leu Val Ile Pro Glu Trp Glu Ala Glu Val Ser
1040 1045 1050
Gln Glu Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val
1055 1060 1065
Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His
1070 1075 1080
Glu Ile Glu Asp Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val
1085 1090 1095
Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asn Tyr
1100 1105 1110
Thr Gly Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn
1115 1120 1125
Gln Gly Tyr Asp Glu Ala Tyr Gly Asn Asn Pro Ser Val Pro Ala
1130 1135 1140
Asp Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg
1145 1150 1155
Arg Glu Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr
1160 1165 1170
Pro Leu Pro Ala Gly Tyr Val Thr Lys Asp Leu Glu Tyr Phe Pro
1175 1180 1185
Glu Thr Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr
1190 1195 1200
Phe Ile Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1205 1210 1215
<210> 26
<211> 3600
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC868.
<400> 26
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta atcaaatacc tttagtgaaa ggatttagag tttggggggg cacctctgtc 1560
attacaggac caggatttac aggaggggat atccttcgaa gaaatacctt tggtgatttt 1620
gtatctctac aagtcaatat taattcacca attacccaaa gataccgttt aagatttcgt 1680
tacgcttcca gtagggatgc acgagttata gtattaacag gagcggcatc cacaggagtg 1740
ggaggccaag ttagtgtaaa tatgcctctt cagaaaacta tggaaatagg ggagaactta 1800
acatctagaa catttagata taccgatttt agtaatcctt tttcatttag agctaatcca 1860
gatataattg ggataagtga acaacctcta tttggtgcag gttctattag tagcggtgaa 1920
ctttatatag ataaaattga aattattcta gcagatgcaa catttgaagc agaatctgat 1980
ttagaaagag cacaaaaggc ggtgaatgag ctgtttactt cttccaatca aatcgggtta 2040
aaaacagatg tgacggatta tcatattgat caagtatcca atttagttga gtgtttatct 2100
gatgaatttt gtctggatga aaaaaaagaa ttgtccgaga aagtcaaaca tgcgaagcga 2160
cttagtgatg agcggaattt acttcaagat ccaaacttta gagggatcaa tagacaacta 2220
gaccgtggct ggagaggaag tacggatatt accatccaag gaggcgatga cgtattcaaa 2280
gagaattacg ttacgctatt gggtaccttt gatgagtgct atccaacgta tttatatcaa 2340
aaaatagatg agtcgaaatt aaaagcctat acccgttacc aattaagagg gtatatcgaa 2400
gatagtcaag acttagaaat ctatttaatt cgctacaatg ccaaacacga aacagtaaat 2460
gtgccaggta cgggttcctt atggccgctt tcagccccaa gtccaatcgg aaaatgtgcc 2520
catcattccc atcatttctc cttggacatt gatgttggat gtacagactt aaatgaggac 2580
ttaggtgtat gggtgatatt caagattaag acgcaagatg gccatgcaag actaggaaat 2640
ctagaatttc tcgaagagaa accattagta ggagaagcac tagctcgtgt gaaaagagcg 2700
gagaaaaaat ggagagacaa acgtgaaaaa ttggaatggg aaacaaatat tgtttataaa 2760
gaggcaaaag aatctgtaga tgctttattt gtaaactctc aatatgatag attacaagcg 2820
gataccaaca tcgcgatgat tcatgcggca gataaacgcg ttcatagcat tcgagaagct 2880
tatctgcctg agctgtctgt gattccgggt gtcaatgcgg ctatttttga agaattagaa 2940
gggcgtattt tcactgcatt ctccctatat gatgcgagaa atgtcattaa aaatggtgat 3000
tttaataatg gcttatcctg ctggaacgtg aaagggcatg tagatgtaga agaacaaaac 3060
aaccaccgtt cggtccttgt tgttccggaa tgggaagcag aagtgtcaca agaagttcgt 3120
gtctgtccgg gtcgtggcta tatccttcgt gtcacagcgt acaaggaggg atatggagaa 3180
ggttgcgtaa ccattcatga gatcgagaac aatacagacg aactgaagtt tagcaactgt 3240
gtagaagagg aagtatatcc aaacaacacg gtaacgtgta atgattatac tgcgactcaa 3300
gaagaatatg agggtacgta cacttctcgt aatcgaggat atgacggagc ctatgaaagc 3360
aattcttctg taccagctga ttatgcatca gcctatgaag aaaaagcata tacagatgga 3420
cgaagagaca atccttgtga atctaacaga ggatatgggg attacacacc actaccagct 3480
ggctatgtga caaaagaatt agagtacttc ccagaaaccg ataaggtatg gattgagatc 3540
ggagaaacgg aaggaacatt catcgtggac agcgtggaat tacttcttat ggaggaatag 3600
<210> 27
<211> 3600
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868.
<400> 27
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgttcgaggc cgagtctgac 1980
ctggagcggg ctcagaaggc tgtcaacgaa ctgttcacca gcagcaacca gattgggctc 2040
aagaccgacg tcacggacta tcacattgac caagtgtcca accttgtgga gtgcctgtcc 2100
gacgagttct gcctcgacga gaagaaggag ctgtccgaga aggtcaaaca cgcgaagcgt 2160
ctgagtgacg agcggaattt gctccaggac ccgaacttcc gtggcatcaa ccgccagctc 2220
gaccgtggtt ggcgcgggag tacagacatc accatccagg gaggcgacga tgtgttcaag 2280
gagaactatg tgacgctgct cgggactttc gacgaatgct acccgacgta tctctaccag 2340
aagatagacg agagtaaatt gaaggcgtac acccgctacc agcttcgcgg gtacatcgag 2400
gatagtcagg acctggaaat ctacctgatc cgatacaacg ccaagcacga gacagtgaac 2460
gtgccaggca cgggctcact ttggccattg agcgctccct ctccaatcgg aaagtgcgct 2520
caccactcgc accacttctc tctggacatc gacgtgggct gcaccgacct caacgaggac 2580
ctgggtgtct gggttatctt caagattaag acccaggacg gacatgcccg cctcggcaac 2640
ctggagttcc ttgaggagaa gcctctcgtg ggcgaggccc tcgctcgtgt gaagcgcgcc 2700
gagaagaaat ggcgagacaa gcgggagaag ctggagtggg agaccaacat cgtgtacaag 2760
gaggccaagg agtcagtgga cgcactcttc gtcaacagcc agtacgaccg cctccaggct 2820
gacaccaaca tcgccatgat ccacgcggct gacaagcggg tccacagcat ccgtgaggcg 2880
tacctgcccg agctgtcagt gatccctggt gtgaacgcgg cgatcttcga ggaactggag 2940
ggccgcatct tcacagcatt cagcctgtac gatgccagga atgttattaa gaacggtgac 3000
ttcaacaacg ggctgagttg ctggaacgtc aagggccatg tggacgtcga ggagcagaac 3060
aaccaccggt ccgtgctggt cgtgccggag tgggaggcag aggtgagcca ggaggtccgc 3120
gtctgccctg gtcgcggcta catcctccgt gtgactgcgt acaaggaagg ctacggtgaa 3180
ggctgcgtga ctatccacga gatcgagaac aacaccgacg agctcaagtt ctcgaactgt 3240
gtggaggagg aggtgtaccc gaacaacacc gttacttgca acgactacac tgccacgcaa 3300
gaggagtacg agggcactta cacttcccgg aatcgcggct atgatggcgc gtacgagtcc 3360
aacagcagcg tgcctgcgga ttatgcgtcc gcttacgagg agaaggcgta caccgacgga 3420
cggagggaca acccttgcga gtccaaccgt ggctacggtg actacactcc gctgcccgcc 3480
gggtacgtca ccaaggagct ggagtacttc ccggagaccg acaaagtctg gatcgagatc 3540
ggcgagacgg agggcacttt catcgtggac tcggtcgagc tgctactgat ggaggagtga 3600
<210> 28
<211> 1199
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein TIC868.
<400> 28
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu
645 650 655
Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Glu Leu Phe
660 665 670
Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His
675 680 685
Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser Asp Glu Phe Cys
690 695 700
Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg
705 710 715 720
Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile
725 730 735
Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile
740 745 750
Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Leu Gly
755 760 765
Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu
770 775 780
Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu
785 790 795 800
Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His
805 810 815
Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala
820 825 830
Pro Ser Pro Ile Gly Lys Cys Ala His His Ser His His Phe Ser Leu
835 840 845
Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp
850 855 860
Val Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn
865 870 875 880
Leu Glu Phe Leu Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg
885 890 895
Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu
900 905 910
Trp Glu Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala
915 920 925
Leu Phe Val Asn Ser Gln Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile
930 935 940
Ala Met Ile His Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala
945 950 955 960
Tyr Leu Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe
965 970 975
Glu Glu Leu Glu Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala
980 985 990
Arg Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp
995 1000 1005
Asn Val Lys Gly His Val Asp Val Glu Glu Gln Asn Asn His Arg
1010 1015 1020
Ser Val Leu Val Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu
1025 1030 1035
Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala
1040 1045 1050
Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile
1055 1060 1065
Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val Glu Glu
1070 1075 1080
Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp Tyr Thr Ala
1085 1090 1095
Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Arg Gly
1100 1105 1110
Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala Asp Tyr
1115 1120 1125
Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg Asp
1130 1135 1140
Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu
1145 1150 1155
Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr
1160 1165 1170
Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile
1175 1180 1185
Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1190 1195
<210> 29
<211> 3600
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_9.
<400> 29
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
agcctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgcag 1200
gcgggcatta acatccttat gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgaagaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgttcgaggc cgagtctgac 1980
ctggagcggg ctcagaaggc tgtcaacgaa ctgttcacca gcagcaacca gattgggctc 2040
aagaccgacg tcacggacta tcacattgac caagtgtcca accttgtgga gtgcctgtcc 2100
gacgagttct gcctcgacga gaagaaggag ctgtccgaga aggtcaaaca cgcgaagcgt 2160
ctgagtgacg agcggaattt gctccaggac ccgaacttcc gtggcatcaa ccgccagctc 2220
gaccgtggtt ggcgcgggag tacagacatc accatccagg gaggcgacga tgtgttcaag 2280
gagaactatg tgacgctgct cgggactttc gacgaatgct acccgacgta tctctaccag 2340
aagatagacg agagtaaatt gaaggcgtac acccgctacc agcttcgcgg gtacatcgag 2400
gatagtcagg acctggaaat ctacctgatc cgatacaacg ccaagcacga gacagtgaac 2460
gtgccaggca cgggctcact ttggccattg agcgctccct ctccaatcgg aaagtgcgct 2520
caccactcgc accacttctc tctggacatc gacgtgggct gcaccgacct caacgaggac 2580
ctgggtgtct gggttatctt caagattaag acccaggacg gacatgcccg cctcggcaac 2640
ctggagttcc ttgaggagaa gcctctcgtg ggcgaggccc tcgctcgtgt gaagcgcgcc 2700
gagaagaaat ggcgagacaa gcgggagaag ctggagtggg agaccaacat cgtgtacaag 2760
gaggccaagg agtcagtgga cgcactcttc gtcaacagcc agtacgaccg cctccaggct 2820
gacaccaaca tcgccatgat ccacgcggct gacaagcggg tccacagcat ccgtgaggcg 2880
tacctgcccg agctgtcagt gatccctggt gtgaacgcgg cgatcttcga ggaactggag 2940
ggccgcatct tcacagcatt cagcctgtac gatgccagga atgttattaa gaacggtgac 3000
ttcaacaacg ggctgagttg ctggaacgtc aagggccatg tggacgtcga ggagcagaac 3060
aaccaccggt ccgtgctggt cgtgccggag tgggaggcag aggtgagcca ggaggtccgc 3120
gtctgccctg gtcgcggcta catcctccgt gtgactgcgt acaaggaagg ctacggtgaa 3180
ggctgcgtga ctatccacga gatcgagaac aacaccgacg agctcaagtt ctcgaactgt 3240
gtggaggagg aggtgtaccc gaacaacacc gttacttgca acgactacac tgccacgcaa 3300
gaggagtacg agggcactta cacttcccgg aatcgcggct atgatggcgc gtacgagtcc 3360
aacagcagcg tgcctgcgga ttatgcgtcc gcttacgagg agaaggcgta caccgacgga 3420
cggagggaca acccttgcga gtccaaccgt ggctacggtg actacactcc gctgcccgcc 3480
gggtacgtca ccaaggagct ggagtacttc ccggagaccg acaaagtctg gatcgagatc 3540
ggcgagacgg agggcacttt catcgtggac tcggtcgagc tgctactgat ggaggagtga 3600
<210> 30
<211> 1199
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_9.
<400> 30
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Ser Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Gln
385 390 395 400
Ala Gly Ile Asn Ile Leu Met Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Lys Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu
645 650 655
Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Glu Leu Phe
660 665 670
Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His
675 680 685
Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser Asp Glu Phe Cys
690 695 700
Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg
705 710 715 720
Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile
725 730 735
Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile
740 745 750
Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Leu Gly
755 760 765
Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu
770 775 780
Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu
785 790 795 800
Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His
805 810 815
Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala
820 825 830
Pro Ser Pro Ile Gly Lys Cys Ala His His Ser His His Phe Ser Leu
835 840 845
Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp
850 855 860
Val Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn
865 870 875 880
Leu Glu Phe Leu Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg
885 890 895
Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu
900 905 910
Trp Glu Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala
915 920 925
Leu Phe Val Asn Ser Gln Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile
930 935 940
Ala Met Ile His Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala
945 950 955 960
Tyr Leu Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe
965 970 975
Glu Glu Leu Glu Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala
980 985 990
Arg Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp
995 1000 1005
Asn Val Lys Gly His Val Asp Val Glu Glu Gln Asn Asn His Arg
1010 1015 1020
Ser Val Leu Val Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu
1025 1030 1035
Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala
1040 1045 1050
Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile
1055 1060 1065
Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val Glu Glu
1070 1075 1080
Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp Tyr Thr Ala
1085 1090 1095
Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Arg Gly
1100 1105 1110
Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala Asp Tyr
1115 1120 1125
Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg Asp
1130 1135 1140
Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu
1145 1150 1155
Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr
1160 1165 1170
Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile
1175 1180 1185
Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1190 1195
<210> 31
<211> 3678
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC868_10.
<400> 31
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta atcaaatacc tttagtgaaa ggatttagag tttggggggg cacctctgtc 1560
attacaggac caggatttac aggaggggat atccttcgaa gaaatacctt tggtgatttt 1620
gtatctctac aagtcaatat taattcacca attacccaaa gataccgttt aagatttcgt 1680
tacgcttcca gtagggatgc acgagttata gtattaacag gagcggcatc cacaggagtg 1740
ggaggccaag ttagtgtaaa tatgcctctt cagaaaacta tggaaatagg ggagaactta 1800
acatctagaa catttagata taccgatttt agtaatcctt tttcatttag agctaatcca 1860
gatataattg ggataagtga acaacctcta tttggtgcag gttctattag tagcggtgaa 1920
ctttatatag ataaaattga aattattcta gcagatgcaa catttgaggc agaatatgat 1980
ttagaaagag cgcaaaaggt ggtgaatgcc ctgtttacgt ctacaaacca actagggcta 2040
aaaacagatg tgacggatta tcatattgat caggtatcca atctagttgc gtgtttatcg 2100
gatgaatttt gtctggatga aaagagagaa ttgtccgaga aagttaaaca tgcaaagcga 2160
ctcagtgatg agcggaattt acttcaagat ccaaacttca gagggatcaa taggcaacca 2220
gaccgtggct ggagaggaag tacggatatt actatccaag gaggagatga cgtattcaaa 2280
gagaattacg ttacgctacc gggtaccttt gatgagtgct atccaacgta tttatatcaa 2340
aaaatagatg agtcgaaatt aaaagcctat acccgttatc aattaagagg gtatatcgaa 2400
gatagtcaag acttagaaat ctatttaatt cgttacaatg caaaacacga aatagtaaat 2460
gtaccaggta caggaagttt atggcctctt tctgtagaaa atcaaattgg accttgtgga 2520
gaaccgaatc gatgcgcgcc acaccttgaa tggaatcctg atttacactg ttcctgcaga 2580
gacggggaaa aatgtgcaca tcattctcat catttctctt tggacattga tgttggatgt 2640
acagacttaa atgaggactt aggtgtatgg gtgatattca agattaagac gcaagatggc 2700
cacgcacgac tagggaatct agagtttctc gaagagaaac cattattagg agaagcacta 2760
gctcgtgtga aaagagcgga gaaaaaatgg agagacaaac gcgaaacatt acaattggaa 2820
acaactatcg tttataaaga ggcaaaagaa tctgtagatg ctttatttgt aaactctcaa 2880
tatgatagat tacaagcgga tacgaacatc gcgatgattc atgcggcaga taaacgcgtt 2940
catagaattc gagaagcgta tctgccggag ctgtctgtga ttccgggtgt caatgcggct 3000
atttttgaag aattagaaga gcgtattttc actgcatttt ccctatatga tgcgagaaat 3060
attattaaaa atggcgattt caataatggc ttattatgct ggaacgtgaa agggcatgta 3120
gaggtagaag aacaaaacaa tcaccgttca gtcctggtta tcccagaatg ggaggcagaa 3180
gtgtcacaag aggttcgtgt ctgtccaggt cgtggctata tccttcgtgt tacagcgtac 3240
aaagagggat atggagaagg ttgcgtaacg atccatgaga tcgagaacaa tacagacgaa 3300
ctgaaattca acaactgtgt agaagaggaa gtatatccaa acaacacggt aacgtgtatt 3360
aattatactg cgactcaaga agaatatgag ggtacgtaca cttctcgtaa tcgaggatat 3420
gacgaagcct atggtaataa cccttccgta ccagctgatt atgcgtcagt ctatgaagaa 3480
aaatcgtata cagatagacg aagagagaat ccttgtgaat ctaacagagg atatggagat 3540
tacacaccac taccagctgg ttatgtaaca aaggaattag agtacttccc agagaccgat 3600
aaggtatgga ttgagattgg agaaacagaa ggaacattca tcgtggacag cgtggaatta 3660
ctccttatgg aggaatag 3678
<210> 32
<211> 3678
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_10.
<400> 32
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgttcgaggc cgagtacgac 1980
cttgagcgcg cccagaaggt ggtgaacgcc ctcttcacta gcactaacca gctaggcctg 2040
aagactgacg tgaccgacta ccacatcgac caagtgagca acctagtggc ctgcctctcc 2100
gacgagttct gcctcgacga gaagcgcgag ctgtccgaga aggtgaagca cgccaagcgc 2160
ctctccgacg agcgcaacct gctccaggac cccaacttca ggggcatcaa caggcagccc 2220
gaccgcggct ggcgcggctc caccgacatc accatccagg gcggtgacga cgtattcaag 2280
gagaactacg ttaccctccc cggcaccttc gacgagtgtt accccaccta cctctaccag 2340
aagatcgacg agtccaagct gaaggcctac acccgctacc agctccgcgg ctacatcgag 2400
gactcccagg acctggaaat ctacctcatc cgctacaacg ccaagcacga gatcgtgaac 2460
gtgcctggca ccggcagcct ctggcctctc agcgtggaga accagatcgg cccttgcggc 2520
gagcctaacc gctgcgcccc tcacctcgag tggaaccctg acctccactg ctcgtgcagg 2580
gacggcgaga agtgcgccca ccatagccac cacttctctc tggacatcga cgtgggctgc 2640
accgacctga acgaggacct gggcgtgtgg gttatcttca agatcaagac ccaggacggt 2700
cacgccaggc tgggtaacct ggagttcctt gaggaaaagc ctctgctggg tgaggccctg 2760
gccagggtca agagggctga gaagaaatgg agggataaga gggagaccct gcagctggag 2820
accactatcg tctacaagga ggctaaggag tctgtcgatg ctctgttcgt caactctcag 2880
tacgatagac tgcaagctga taccaacatc gctatgatcc acgctgcgga taagcgggtc 2940
caccggatcc gggaggctta ccttccggag ctttctgtca tcccgggtgt caacgctgcg 3000
atcttcgagg aacttgagga acggatcttc actgcgttta gtctttacga tgcgcggaac 3060
atcatcaaga acggggactt caacaatggt ctgctgtgct ggaacgtcaa gggtcatgtc 3120
gaggtcgagg aacaaaacaa tcatcgtagt gtccttgtca ttcctgagtg ggaggcggag 3180
gtctctcaag aggtccgtgt ttgcccgggg cgtgggtaca ttcttcgtgt tactgcgtac 3240
aaggaggggt acggggaggg gtgcgttact attcatgaga ttgagaacaa tactgatgag 3300
cttaagttca acaattgtgt tgaggaggag gtttacccga acaatactgt tacgtgcatc 3360
aactacacgg caacgcaaga ggaatacgag gggacgtaca cctcgcgtaa tagagggtat 3420
gatgaggcgt acggaaacaa cccgtcggtt ccagcagatt atgcctcggt ttatgaggag 3480
aagtcgtaca cggatagacg acgcgagaat ccatgtgagt caaatcgagg atacggagat 3540
tacacaccat taccagcagg atacgttaca aaggagttgg aatacttccc ggaaacagat 3600
aaagtttgga ttgaaatcgg agaaacagaa ggaacattca tcgtcgactc agtagaattg 3660
ttgttgatgg aagaatga 3678
<210> 33
<211> 1225
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_10.
<400> 33
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu
645 650 655
Ala Glu Tyr Asp Leu Glu Arg Ala Gln Lys Val Val Asn Ala Leu Phe
660 665 670
Thr Ser Thr Asn Gln Leu Gly Leu Lys Thr Asp Val Thr Asp Tyr His
675 680 685
Ile Asp Gln Val Ser Asn Leu Val Ala Cys Leu Ser Asp Glu Phe Cys
690 695 700
Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg
705 710 715 720
Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile
725 730 735
Asn Arg Gln Pro Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile
740 745 750
Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Pro Gly
755 760 765
Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu
770 775 780
Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu
785 790 795 800
Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His
805 810 815
Glu Ile Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Val
820 825 830
Glu Asn Gln Ile Gly Pro Cys Gly Glu Pro Asn Arg Cys Ala Pro His
835 840 845
Leu Glu Trp Asn Pro Asp Leu His Cys Ser Cys Arg Asp Gly Glu Lys
850 855 860
Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys
865 870 875 880
Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys
885 890 895
Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu
900 905 910
Lys Pro Leu Leu Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys
915 920 925
Lys Trp Arg Asp Lys Arg Glu Thr Leu Gln Leu Glu Thr Thr Ile Val
930 935 940
Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln
945 950 955 960
Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala
965 970 975
Asp Lys Arg Val His Arg Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser
980 985 990
Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Glu Arg
995 1000 1005
Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Ile Ile Lys
1010 1015 1020
Asn Gly Asp Phe Asn Asn Gly Leu Leu Cys Trp Asn Val Lys Gly
1025 1030 1035
His Val Glu Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val
1040 1045 1050
Ile Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys
1055 1060 1065
Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly
1070 1075 1080
Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile Glu Asn Asn Thr
1085 1090 1095
Asp Glu Leu Lys Phe Asn Asn Cys Val Glu Glu Glu Val Tyr Pro
1100 1105 1110
Asn Asn Thr Val Thr Cys Ile Asn Tyr Thr Ala Thr Gln Glu Glu
1115 1120 1125
Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Arg Gly Tyr Asp Glu Ala
1130 1135 1140
Tyr Gly Asn Asn Pro Ser Val Pro Ala Asp Tyr Ala Ser Val Tyr
1145 1150 1155
Glu Glu Lys Ser Tyr Thr Asp Arg Arg Arg Glu Asn Pro Cys Glu
1160 1165 1170
Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu Pro Ala Gly Tyr
1175 1180 1185
Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp Lys Val Trp
1190 1195 1200
Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp Ser Val
1205 1210 1215
Glu Leu Leu Leu Met Glu Glu
1220 1225
<210> 34
<211> 3726
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC868_11.
<400> 34
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta atcaaatacc tttagtgaaa ggatttagag tttggggggg cacctctgtc 1560
attacaggac caggatttac aggaggggat atccttcgaa gaaatacctt tggtgatttt 1620
gtatctctac aagtcaatat taattcacca attacccaaa gataccgttt aagatttcgt 1680
tacgcttcca gtagggatgc acgagttata gtattaacag gagcggcatc cacaggagtg 1740
ggaggccaag ttagtgtaaa tatgcctctt cagaaaacta tggaaatagg ggagaactta 1800
acatctagaa catttagata taccgatttt agtaatcctt tttcatttag agctaatcca 1860
gatataattg ggataagtga acaacctcta tttggtgcag gttctattag tagcggtgaa 1920
ctttatatag ataaaattga aattattcta gcagatgcaa caggaacgac aacctatgag 1980
tatgaagaga agcagaatct agaaaaagcg cagaaagcgt tgaacgcttt gtttacggat 2040
ggcacgaatg gctatctaca aatggatgcc actgattatg atatcaatca aactgcaaac 2100
ttaatagaat gtgtatcaga tgaattgtat gcaaaagaaa agatagtttt attagatgaa 2160
gtcaaatatg cgaagcggct tagcatatca cgtaacctac ttttgaacga tgatttagaa 2220
ttttcagatg gatttggaga aaacggatgg acgacaagtg ataatatttc aatccaggcg 2280
gataatcccc tttttaaggg gaattattta aaaatgtttg gggcaagaga tattgatgga 2340
accctatttc caacttatct ctatcaaaaa atagatgagt ccaggttaaa accatataca 2400
cgttatcgag taagagggtt tgtgggaagt agtaaaaatc taaaattagt ggtaacacgc 2460
tatgagaaag aaattgatgc cattatgaat gttccaaatg atttggcaca tatgcagctt 2520
aacccttcat gtggagatta tcgctgtgaa tcatcgtccc agtttttggt gaaccaagtg 2580
catcctacac caacagctgg atatgctctt gatatgtatg catgcccgtc aagttcagat 2640
aaaaaacata ttatgtgtca cgatcgtcat ccatttgatt ttcatattga caccggagaa 2700
ttaaatccaa acacaaacct gggtattgat gtcttgttta aaatttctaa tccaaatgga 2760
tacgctacat tagggaatct agaagtcatt gaagaaggac cactaacaga tgaagcattg 2820
gtacatgtaa aacaaaagga aaagaaatgg cgtcagcaca tggagaaaaa acgaatggaa 2880
acacaacaag cctatgatcc agcaaaacaa gctgtagatg cattatttac aaatgaacaa 2940
gagttagact atcatactac tttagatcat attcagaacg ccgatcagct ggtacaggcg 3000
attccctatg tacaccatgc ttggttaccg gatgctccag gtatgaacta tgatgtatat 3060
caagggttaa acgcacgtat catgcaggcg tacaatttat atgatgcacg aaatgtcata 3120
ataaatggtg actttacaca aggactacaa ggatggcacg caacaggaaa agcagcggta 3180
caacaaatag atggagcttc agtattagtt ctatcaaact ggagtgccga ggtatctcag 3240
aatctgcatg cccaagatca tcatggatat atgttacgtg tgattgccaa aaaagaaggt 3300
cctggaaaag ggtatgtaat gatgatggat tttaatggaa agcaggaaac acttacgttc 3360
acttcttgtg aagaaggata tataacaaaa acaatagagg tattcccgga aagtgatcga 3420
atacgaattg aaatgggaga aacagagggt acgttttatg tagatagcat cgagttgctt 3480
tgtatgcaag gatatgctag cgataataac ccgcacacgg gtaatatgta tgagcaaagt 3540
tataatggaa attataatca aaatactagc gatgtgtatc accaaggata tataaacaac 3600
tataaccaaa attctagtag tatgtataat caaaattata ttaacaatga tgacctgcat 3660
tccggttgca catgtaacca agggcataac tctggctgta catgtaatca aggatataac 3720
cgttag 3726
<210> 35
<211> 3726
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_11.
<400> 35
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cggggactac cacctacgag 1980
tacgaggaga agcagaatct cgagaaggct cagaaggctc tgaacgctct gttcactgac 2040
gggaccaacg gctacctcca gatggacgcc actgactacg acatcaacca gacagctaac 2100
ctgattgagt gtgtgagtga cgaactgtac gctaaggaga agatcgtact cctggacgag 2160
gtgaagtacg ctaagcgcct gagcattagc cgtaacctgc tgctgaacga cgatctggag 2220
ttcagcgacg gctttggcga gaacggctgg accaccagcg acaacatctc catccaggcc 2280
gacaatccac tcttcaaagg caactacctc aagatgttcg gagccaggga catcgacggc 2340
accctctttc cgacctacct ctaccagaag atcgacgagt cccgcctcaa accctacacc 2400
cgctacaggg tgcgcggctt cgtgggcagc agcaagaacc tcaagctcgt ggtcacacgg 2460
tatgagaagg agatcgacgc catcatgaac gtgcccaacg atctcgccca catgcagctc 2520
aatccatcct gcggcgacta ccggtgcgag tccagctccc agttcctcgt gaaccaggtg 2580
caccctactc cgaccgctgg ctatgccctg gacatgtacg cctgccctag ttcctccgac 2640
aagaagcaca tcatgtgcca cgaccgtcat ccgttcgact tccacatcga caccggcgaa 2700
ctgaacccga acaccaacct gggcatcgac gtactgttca agatttccaa cccgaacggg 2760
tacgccacct tgggcaacct ggaggtcatc gaagaaggcc cgctgaccga cgaggccctg 2820
gtccacgtca aacagaagga gaagaagtgg cggcagcaca tggagaagaa gcggatggag 2880
actcaacaag cctacgaccc ggccaagcaa gctgtggacg ctctgttcac caacgagcaa 2940
gagcttgact accacactac tcttgaccac atccagaatg ctgaccagct tgtccaggct 3000
attccgtacg tccaccacgc ttggctaccg gacgctccag ggatgaacta cgatgtgtac 3060
cagggtctga acgcgcggat catgcaagcg tacaacctgt acgacgcgcg taacgtcatc 3120
atcaacggtg acttcactca gggtcttcaa ggttggcacg cgactggcaa agcggcagtc 3180
cagcagattg atggtgcgtc tgttcttgtg ttgagcaact ggtctgcgga ggtttctcag 3240
aacctgcacg cacaggatca ccacggctac atgctgaggg tgattgctaa gaaggagggc 3300
cctggcaaag gctacgtcat gatgatggac ttcaacggaa agcaagaaac cctgaccttc 3360
actagctgtg aggagggcta catcactaag accattgagg tctttccgga gtctgaccgc 3420
atccggatcg agatgggcga gaccgaaggc acgttctacg tggactccat cgaactcctc 3480
tgcatgcaag gctacgcctc cgacaacaac ccacacacgg gcaacatgta cgagcagtcc 3540
tacaacggga actacaacca gaacacctcc gatgtgtacc atcagggcta catcaacaac 3600
tacaaccaga acagcagcag catgtacaac cagaactaca tcaacaacga tgacttgcac 3660
tcgggttgca cctgcaacca gggtcacaac agtgggtgca cgtgcaacca gggatacaac 3720
cgttga 3726
<210> 36
<211> 1241
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_11.
<400> 36
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Gly Thr
645 650 655
Thr Thr Tyr Glu Tyr Glu Glu Lys Gln Asn Leu Glu Lys Ala Gln Lys
660 665 670
Ala Leu Asn Ala Leu Phe Thr Asp Gly Thr Asn Gly Tyr Leu Gln Met
675 680 685
Asp Ala Thr Asp Tyr Asp Ile Asn Gln Thr Ala Asn Leu Ile Glu Cys
690 695 700
Val Ser Asp Glu Leu Tyr Ala Lys Glu Lys Ile Val Leu Leu Asp Glu
705 710 715 720
Val Lys Tyr Ala Lys Arg Leu Ser Ile Ser Arg Asn Leu Leu Leu Asn
725 730 735
Asp Asp Leu Glu Phe Ser Asp Gly Phe Gly Glu Asn Gly Trp Thr Thr
740 745 750
Ser Asp Asn Ile Ser Ile Gln Ala Asp Asn Pro Leu Phe Lys Gly Asn
755 760 765
Tyr Leu Lys Met Phe Gly Ala Arg Asp Ile Asp Gly Thr Leu Phe Pro
770 775 780
Thr Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Arg Leu Lys Pro Tyr Thr
785 790 795 800
Arg Tyr Arg Val Arg Gly Phe Val Gly Ser Ser Lys Asn Leu Lys Leu
805 810 815
Val Val Thr Arg Tyr Glu Lys Glu Ile Asp Ala Ile Met Asn Val Pro
820 825 830
Asn Asp Leu Ala His Met Gln Leu Asn Pro Ser Cys Gly Asp Tyr Arg
835 840 845
Cys Glu Ser Ser Ser Gln Phe Leu Val Asn Gln Val His Pro Thr Pro
850 855 860
Thr Ala Gly Tyr Ala Leu Asp Met Tyr Ala Cys Pro Ser Ser Ser Asp
865 870 875 880
Lys Lys His Ile Met Cys His Asp Arg His Pro Phe Asp Phe His Ile
885 890 895
Asp Thr Gly Glu Leu Asn Pro Asn Thr Asn Leu Gly Ile Asp Val Leu
900 905 910
Phe Lys Ile Ser Asn Pro Asn Gly Tyr Ala Thr Leu Gly Asn Leu Glu
915 920 925
Val Ile Glu Glu Gly Pro Leu Thr Asp Glu Ala Leu Val His Val Lys
930 935 940
Gln Lys Glu Lys Lys Trp Arg Gln His Met Glu Lys Lys Arg Met Glu
945 950 955 960
Thr Gln Gln Ala Tyr Asp Pro Ala Lys Gln Ala Val Asp Ala Leu Phe
965 970 975
Thr Asn Glu Gln Glu Leu Asp Tyr His Thr Thr Leu Asp His Ile Gln
980 985 990
Asn Ala Asp Gln Leu Val Gln Ala Ile Pro Tyr Val His His Ala Trp
995 1000 1005
Leu Pro Asp Ala Pro Gly Met Asn Tyr Asp Val Tyr Gln Gly Leu
1010 1015 1020
Asn Ala Arg Ile Met Gln Ala Tyr Asn Leu Tyr Asp Ala Arg Asn
1025 1030 1035
Val Ile Ile Asn Gly Asp Phe Thr Gln Gly Leu Gln Gly Trp His
1040 1045 1050
Ala Thr Gly Lys Ala Ala Val Gln Gln Ile Asp Gly Ala Ser Val
1055 1060 1065
Leu Val Leu Ser Asn Trp Ser Ala Glu Val Ser Gln Asn Leu His
1070 1075 1080
Ala Gln Asp His His Gly Tyr Met Leu Arg Val Ile Ala Lys Lys
1085 1090 1095
Glu Gly Pro Gly Lys Gly Tyr Val Met Met Met Asp Phe Asn Gly
1100 1105 1110
Lys Gln Glu Thr Leu Thr Phe Thr Ser Cys Glu Glu Gly Tyr Ile
1115 1120 1125
Thr Lys Thr Ile Glu Val Phe Pro Glu Ser Asp Arg Ile Arg Ile
1130 1135 1140
Glu Met Gly Glu Thr Glu Gly Thr Phe Tyr Val Asp Ser Ile Glu
1145 1150 1155
Leu Leu Cys Met Gln Gly Tyr Ala Ser Asp Asn Asn Pro His Thr
1160 1165 1170
Gly Asn Met Tyr Glu Gln Ser Tyr Asn Gly Asn Tyr Asn Gln Asn
1175 1180 1185
Thr Ser Asp Val Tyr His Gln Gly Tyr Ile Asn Asn Tyr Asn Gln
1190 1195 1200
Asn Ser Ser Ser Met Tyr Asn Gln Asn Tyr Ile Asn Asn Asp Asp
1205 1210 1215
Leu His Ser Gly Cys Thr Cys Asn Gln Gly His Asn Ser Gly Cys
1220 1225 1230
Thr Cys Asn Gln Gly Tyr Asn Arg
1235 1240
<210> 37
<211> 3468
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC868_12.
<400> 37
atgacttcaa ataggaaaaa tgagaatgaa attataaatg ctttatcgat tccagctgta 60
tcgaatcatt ccgcacaaat gaatctatca accgatgctc gtattgagga tagcttgtgt 120
atagccgagg ggaacaatat cgatccattt gttagcgcat caacagtcca aacgggtatt 180
aacatagctg gtagaatact aggtgtatta ggcgtaccgt ttgctggaca aatagctagt 240
ttttatagtt ttcttgttgg tgaattatgg ccccgcggca gagatccttg ggaaattttc 300
ctagaacatg tcgaacaact tataagacaa caagtaacag aaaatactag ggatacggct 360
cttgctcgat tacaaggttt aggaaattcc tttagagcct atcaacagtc acttgaagat 420
tggctagaaa accgtgatga tgcaagaacg agaagtgttc tttataccca atatatagcc 480
ttagaacttg attttcttaa tgcgatgccg cttttcgcaa ttagaaacca agaagttcca 540
ttattaatgg tatatgctca agctgcaaat ttacacctat tattattgag agatgcctct 600
ctttttggta gtgaatttgg gcttacatcc caagaaattc aacgttatta tgagcgccaa 660
gtggaaaaaa cgagagaata ttctgattat tgcgcaagat ggtataatac gggtttaaat 720
aatttgagag ggacaaatgc tgaaagttgg ttgcgatata atcaattccg tagagactta 780
acgctaggag tattagatct agtggcacta ttcccaagct atgacacgcg tgtttatcca 840
atgaatacca gtgctcaatt aacaagagaa atttatacag atccaattgg gagaacaaat 900
gcaccttcag gatttgcaag tacgaattgg tttaataata atgcaccatc gttttctgcc 960
atagaggctg ccgttattag gcctccgcat ctacttgatt ttccagaaca gcttacaatt 1020
ttcagcgtat taagtcgatg gagtaatact caatatatga attactgggt gggacataga 1080
cttgaatcgc gaacaataag ggggtcatta agtacctcga cacacggaaa taccaatact 1140
tctattaatc ctgtaacatt acagttcaca tctcgagacg tttatagaac agaatcattt 1200
gcagggataa atatacttct aactactcct gtgaatggag taccttgggc tagatttaat 1260
tggagaaatc ccctgaattc tcttagaggt agccttctct atactatagg gtatactgga 1320
gtggggacac aactatttga ttcagaaact gaattaccac cagaaacaac agaacgacca 1380
aattatgaat cttacagtca tagattatct aatataagac taatatcagg aaacactttg 1440
agagcaccag tatattcttg gacgcaccgt agtgcagatc gtacaaatac cattagttca 1500
gatagcatta atcaaatacc tttagtgaaa ggatttagag tttggggggg cacctctgtc 1560
attacaggac caggatttac aggaggggat atccttcgaa gaaatacctt tggtgatttt 1620
gtatctctac aagtcaatat taattcacca attacccaaa gataccgttt aagatttcgt 1680
tacgcttcca gtagggatgc acgagttata gtattaacag gagcggcatc cacaggagtg 1740
ggaggccaag ttagtgtaaa tatgcctctt cagaaaacta tggaaatagg ggagaactta 1800
acatctagaa catttagata taccgatttt agtaatcctt tttcatttag agctaatcca 1860
gatataattg ggataagtga acaacctcta tttggtgcag gttctattag tagcggtgaa 1920
ctttatatag ataaaattga aattattcta gcagatgcaa caaatccgac gcgagaggcg 1980
gaagaggatc tagaagcagc gaagaaagcg gtggcgagct tgtttacacg tacaagggac 2040
ggattacaag taaatgtgac agattatcaa gtcgatcaag cggcaaattt agtgtcatgc 2100
ttatcagatg aacaatatgg gcatgacaaa aagatgttat tggaagcggt aagagcggca 2160
aaacgcctca gccgagaacg caacttactt caggatccag attttaatac aatcaatagt 2220
acagaagaaa atggatggaa agcaagtaac ggcgttacta ttagcgaggg cggtccattc 2280
tataaaggcc gtgcgcttca gctagcaagc gcaagagaaa attacccaac atacatttat 2340
caaaaagtaa atgcatcaga gttaaagccg tatacacgtt atagactgga tgggttcgtg 2400
aagagtagtc aagatttaga aattgatctc attcaccatc ataaagtcca tctcgtgaaa 2460
aatgtaccag ataatttagt atccgatact tactcggatg gttcttgcag tggaatgaat 2520
cgatgtgagg aacaacagat ggtaaatgcg caactggaaa cagaacatca tcatccgatg 2580
gattgctgtg aagcggctca aacacatgag ttttcttcct atattaatac aggcgatcta 2640
aattcaagtg tagatcaagg catttgggtt gtattgaaag ttcgaacaac cgatggttat 2700
gcgacgctag gaaatcttga attggtagag gtcggaccgt tatcgggtga atctctagaa 2760
cgtgaacaaa gggataatgc gaaatggagt gcagagctag gaagaaagcg tgcagaaaca 2820
gatcgcgtgt atcaagatgc caaacaatcc atcaatcatt tatttgtgga ttatcaagat 2880
caacaattaa atccagaaat agggatggca gatattattg acgctcaaaa tcttgtcgca 2940
tcaatttcag atgtgtatag cgatgcagta ctgcaaatcc ctggaattaa ctatgagatt 3000
tacacagagc tatccaatcg cttacaacaa gcatcgtatc tgtatacgtc tcgaaatgcg 3060
gtgcaaaatg gggactttaa cagcggtcta gatagttgga atgcaacagg gggggctacg 3120
gtacaacagg atggcaatac gcatttctta gttctttctc attgggatgc acaagtttct 3180
caacaattta gagtgcagcc gaattgtaaa tatgtattac gtgtaacagc agagaaagta 3240
ggcggcggag acggatacgt gacaatccgg gatggtgctc atcatacaga aaagcttaca 3300
tttaatgcat gtgattatga tataaatggc acgtacgtga ctgataatac gtatctaaca 3360
aaagaagtgg tattctattc acatacagaa cacatgtggg tagaggtaag tgaaacagaa 3420
ggtgcatttc atatagatag tattgaattc gttgaaacag aaaagtag 3468
<210> 38
<211> 3468
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_12.
<400> 38
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgaacccgac gcgggaagct 1980
gaggaagact tggaagccgc caagaaagcg gtcgccagcc tgtttactcg gacgcgggac 2040
gggctccaag tgaatgtgac ggactatcaa gtggatcagg ccgctaacct cgtgtcatgc 2100
ctgagcgacg agcagtacgg tcacgacaag aaaatgctgc tggaggccgt ccgggccgcc 2160
aagcggctgt ccagggagcg taacctgcta caagatcccg actttaacac gatcaacagc 2220
acagaggaga atggctggaa ggccagcaac ggagttacga taagcgaggg cggtccgttc 2280
tacaagggtc gtgccctcca gctcgcctct gcaagggaga actatccaac ctacatctat 2340
cagaaggtga acgcatccga gcttaagccc tacacacgct accgcctgga cgggttcgtt 2400
aagtccagtc aagacctaga gatagacctc atccaccacc acaaagtgca tctggtcaag 2460
aacgttcccg ataatctcgt gagcgatacc tactcagacg gctcatgctc tggcatgaac 2520
agatgtgagg agcaacagat ggttaatgct caactcgaaa ccgagcatca tcatcctatg 2580
gattgctgcg aggccgcgca gacccatgag ttcagctctt acatcaacac cggagacctc 2640
aacagtagcg tggatcaggg aatttgggtg gtgcttaaag tgcgtacaac cgacggctac 2700
gccaccctcg gcaaccttga gcttgtcgag gtcggaccac ttagcggcga gtccctggaa 2760
cgtgagcagc gggacaacgc caaatggagc gcagagctag ggcgcaaacg cgcggagacg 2820
gaccgggttt atcaggacgc gaagcagtcc atcaatcacc tcttcgtgga ttatcaggac 2880
cagcagctta atccagagat cggcatggcc gacatcatcg acgcccagaa cctagtagcg 2940
tcgatttccg atgtctattc cgacgccgtg cttcaaatac ctggcatcaa ctacgagatc 3000
tacacagagt tgtccaacag gctccagcaa gcgtcatacc tctacaccag ccgcaacgcc 3060
gtccagaatg gcgacttcaa ttccggacta gactcctgga acgccacggg cggagctacg 3120
gtgcaacaag acggcaacac ccacttcctc gtacttagcc actgggacgc tcaagtgagt 3180
cagcaattcc gggttcagcc gaactgcaag tacgtcctgc gcgtaacggc cgagaaggtt 3240
ggaggcggag acggctacgt taccatccgc gacggcgctc accacaccga gaaactgacg 3300
ttcaacgctt gtgactacga catcaacggc acttacgtga cggacaacac ctacctgacg 3360
aaggaggtgg tgttctattc tcacaccgag cacatgtggg ttgaggtcag cgagaccgag 3420
ggagccttcc acattgacag catcgagttc gtggagactg agaagtga 3468
<210> 39
<211> 1155
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_12.
<400> 39
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Asn Pro
645 650 655
Thr Arg Glu Ala Glu Glu Asp Leu Glu Ala Ala Lys Lys Ala Val Ala
660 665 670
Ser Leu Phe Thr Arg Thr Arg Asp Gly Leu Gln Val Asn Val Thr Asp
675 680 685
Tyr Gln Val Asp Gln Ala Ala Asn Leu Val Ser Cys Leu Ser Asp Glu
690 695 700
Gln Tyr Gly His Asp Lys Lys Met Leu Leu Glu Ala Val Arg Ala Ala
705 710 715 720
Lys Arg Leu Ser Arg Glu Arg Asn Leu Leu Gln Asp Pro Asp Phe Asn
725 730 735
Thr Ile Asn Ser Thr Glu Glu Asn Gly Trp Lys Ala Ser Asn Gly Val
740 745 750
Thr Ile Ser Glu Gly Gly Pro Phe Tyr Lys Gly Arg Ala Leu Gln Leu
755 760 765
Ala Ser Ala Arg Glu Asn Tyr Pro Thr Tyr Ile Tyr Gln Lys Val Asn
770 775 780
Ala Ser Glu Leu Lys Pro Tyr Thr Arg Tyr Arg Leu Asp Gly Phe Val
785 790 795 800
Lys Ser Ser Gln Asp Leu Glu Ile Asp Leu Ile His His His Lys Val
805 810 815
His Leu Val Lys Asn Val Pro Asp Asn Leu Val Ser Asp Thr Tyr Ser
820 825 830
Asp Gly Ser Cys Ser Gly Met Asn Arg Cys Glu Glu Gln Gln Met Val
835 840 845
Asn Ala Gln Leu Glu Thr Glu His His His Pro Met Asp Cys Cys Glu
850 855 860
Ala Ala Gln Thr His Glu Phe Ser Ser Tyr Ile Asn Thr Gly Asp Leu
865 870 875 880
Asn Ser Ser Val Asp Gln Gly Ile Trp Val Val Leu Lys Val Arg Thr
885 890 895
Thr Asp Gly Tyr Ala Thr Leu Gly Asn Leu Glu Leu Val Glu Val Gly
900 905 910
Pro Leu Ser Gly Glu Ser Leu Glu Arg Glu Gln Arg Asp Asn Ala Lys
915 920 925
Trp Ser Ala Glu Leu Gly Arg Lys Arg Ala Glu Thr Asp Arg Val Tyr
930 935 940
Gln Asp Ala Lys Gln Ser Ile Asn His Leu Phe Val Asp Tyr Gln Asp
945 950 955 960
Gln Gln Leu Asn Pro Glu Ile Gly Met Ala Asp Ile Ile Asp Ala Gln
965 970 975
Asn Leu Val Ala Ser Ile Ser Asp Val Tyr Ser Asp Ala Val Leu Gln
980 985 990
Ile Pro Gly Ile Asn Tyr Glu Ile Tyr Thr Glu Leu Ser Asn Arg Leu
995 1000 1005
Gln Gln Ala Ser Tyr Leu Tyr Thr Ser Arg Asn Ala Val Gln Asn
1010 1015 1020
Gly Asp Phe Asn Ser Gly Leu Asp Ser Trp Asn Ala Thr Gly Gly
1025 1030 1035
Ala Thr Val Gln Gln Asp Gly Asn Thr His Phe Leu Val Leu Ser
1040 1045 1050
His Trp Asp Ala Gln Val Ser Gln Gln Phe Arg Val Gln Pro Asn
1055 1060 1065
Cys Lys Tyr Val Leu Arg Val Thr Ala Glu Lys Val Gly Gly Gly
1070 1075 1080
Asp Gly Tyr Val Thr Ile Arg Asp Gly Ala His His Thr Glu Lys
1085 1090 1095
Leu Thr Phe Asn Ala Cys Asp Tyr Asp Ile Asn Gly Thr Tyr Val
1100 1105 1110
Thr Asp Asn Thr Tyr Leu Thr Lys Glu Val Val Phe Tyr Ser His
1115 1120 1125
Thr Glu His Met Trp Val Glu Val Ser Glu Thr Glu Gly Ala Phe
1130 1135 1140
His Ile Asp Ser Ile Glu Phe Val Glu Thr Glu Lys
1145 1150 1155
<210> 40
<211> 3732
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_13.
<400> 40
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgacggcgac cttcgaggcg 1980
gagtatgact tggagcgggc tcaggaggcc gtcaacgcgc tgttcacaaa caccaatcct 2040
cgccgcctca agacgggtgt gactgattac cacattgacg aggtctccaa cttggtcgcg 2100
tgtctgtccg atgagttctg cctggacgag aagcgggaac tgctggagaa ggtcaagtac 2160
gccaagcgcc tctccgacga aaggaacctc ctccaagatc ccaactttac ttccattaac 2220
aagcagccgg acttcatctc caccaacgag cagtccaact tcacctcaat ccacgagcag 2280
tcggagcacg ggtggtgggg cagcgagaac atcaccatcc aagagggcaa cgacgtcttc 2340
aaggagaact acgtgatcct gcccggcacc ttcaacgagt gttacccgac ctatctctac 2400
cagaagattg gcgaagcgga actcaaggct tacacccgtt accaactgag tggctacatt 2460
gaggactcac aagacctgga aatctacctg atccgctaca acgccaagca cgagaccctc 2520
gacgtgcctg gcacggagtc cgtctggccc ttgagcgtgg agtctcctat cggtcgttgc 2580
ggcgagccca atcgctgcgc tccgcacttt gagtggaatc ctgatttgga ttgctcctgc 2640
cgagacggtg agaaatgcgc ccaccactcg caccacttca gcctagacat cgacgtgggc 2700
tgcatcgacc tgcacgagaa cttgggcgtc tgggtcgtgt tcaagatcaa gacacaggag 2760
ggccatgctc ggcttgggaa cctggagttc atcgaggaga agccactgct gggtgaagcc 2820
ttgtcacggg tgaaacgcgc cgagaagaag tggcgggaca aacgggagaa gctccagttg 2880
gagacaaagc gtgtgtacac agaggccaag gaggccgtgg atgccttgtt cgtggacagt 2940
cagtacgaca ggctgcaagc ggacaccaac atcgggatga tccacgcggc tgataagctt 3000
gttcacagaa tccgcgaggc gtacctgtca gagcttagcg tgatcccagg cgtcaacgcc 3060
gaaatcttcg aggaactgga gggccgcatt atcacggcaa tctcacttta tgacgcgagg 3120
aatgtggtca agaacggtga cttcaacaac ggcttggcgt gttggaacgt taaagggcac 3180
gtggatgtac aacagtcaca ccacagaagt gtcttggtca tcccggagtg ggaggcggaa 3240
gtgagccagg ccgtccgggt ctgccctggg cgcggttaca tcctccgcgt gacagcgtac 3300
aaggagggct acggtgaggg ctgcgtgacg atccacgaga ttgagaacaa cacggacgag 3360
cttaagttca agaactgcga ggaggaggaa gtgtacccga cagacaccgg cacctgcaac 3420
gactacaccg cccaccaagg gaccgccgcc tgcaacagcc gcaacgcggg ctatgaagat 3480
gcgtacgagg ttgataccac cgcctcagtg aactacaaac cgacttatga ggaggagaca 3540
tacacggacg tcaggcgcga caaccattgt gagtacgacc gtggctacgt gaactatccg 3600
ccggtgccag cgggctacat gacgaaggag ctagaatact tccctgagac ggacaaggtg 3660
tggattgaaa tcggcgagac cgagggcaag tttatcgtgg attctgtcga gctgctgcta 3720
atggaggagt ag 3732
<210> 41
<211> 1243
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_13.
<400> 41
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Thr Ala
645 650 655
Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg Ala Gln Glu Ala Val Asn
660 665 670
Ala Leu Phe Thr Asn Thr Asn Pro Arg Arg Leu Lys Thr Gly Val Thr
675 680 685
Asp Tyr His Ile Asp Glu Val Ser Asn Leu Val Ala Cys Leu Ser Asp
690 695 700
Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Leu Glu Lys Val Lys Tyr
705 710 715 720
Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe
725 730 735
Thr Ser Ile Asn Lys Gln Pro Asp Phe Ile Ser Thr Asn Glu Gln Ser
740 745 750
Asn Phe Thr Ser Ile His Glu Gln Ser Glu His Gly Trp Trp Gly Ser
755 760 765
Glu Asn Ile Thr Ile Gln Glu Gly Asn Asp Val Phe Lys Glu Asn Tyr
770 775 780
Val Ile Leu Pro Gly Thr Phe Asn Glu Cys Tyr Pro Thr Tyr Leu Tyr
785 790 795 800
Gln Lys Ile Gly Glu Ala Glu Leu Lys Ala Tyr Thr Arg Tyr Gln Leu
805 810 815
Ser Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg
820 825 830
Tyr Asn Ala Lys His Glu Thr Leu Asp Val Pro Gly Thr Glu Ser Val
835 840 845
Trp Pro Leu Ser Val Glu Ser Pro Ile Gly Arg Cys Gly Glu Pro Asn
850 855 860
Arg Cys Ala Pro His Phe Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys
865 870 875 880
Arg Asp Gly Glu Lys Cys Ala His His Ser His His Phe Ser Leu Asp
885 890 895
Ile Asp Val Gly Cys Ile Asp Leu His Glu Asn Leu Gly Val Trp Val
900 905 910
Val Phe Lys Ile Lys Thr Gln Glu Gly His Ala Arg Leu Gly Asn Leu
915 920 925
Glu Phe Ile Glu Glu Lys Pro Leu Leu Gly Glu Ala Leu Ser Arg Val
930 935 940
Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Gln Leu
945 950 955 960
Glu Thr Lys Arg Val Tyr Thr Glu Ala Lys Glu Ala Val Asp Ala Leu
965 970 975
Phe Val Asp Ser Gln Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Gly
980 985 990
Met Ile His Ala Ala Asp Lys Leu Val His Arg Ile Arg Glu Ala Tyr
995 1000 1005
Leu Ser Glu Leu Ser Val Ile Pro Gly Val Asn Ala Glu Ile Phe
1010 1015 1020
Glu Glu Leu Glu Gly Arg Ile Ile Thr Ala Ile Ser Leu Tyr Asp
1025 1030 1035
Ala Arg Asn Val Val Lys Asn Gly Asp Phe Asn Asn Gly Leu Ala
1040 1045 1050
Cys Trp Asn Val Lys Gly His Val Asp Val Gln Gln Ser His His
1055 1060 1065
Arg Ser Val Leu Val Ile Pro Glu Trp Glu Ala Glu Val Ser Gln
1070 1075 1080
Ala Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr
1085 1090 1095
Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu
1100 1105 1110
Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Lys Asn Cys Glu Glu
1115 1120 1125
Glu Glu Val Tyr Pro Thr Asp Thr Gly Thr Cys Asn Asp Tyr Thr
1130 1135 1140
Ala His Gln Gly Thr Ala Ala Cys Asn Ser Arg Asn Ala Gly Tyr
1145 1150 1155
Glu Asp Ala Tyr Glu Val Asp Thr Thr Ala Ser Val Asn Tyr Lys
1160 1165 1170
Pro Thr Tyr Glu Glu Glu Thr Tyr Thr Asp Val Arg Arg Asp Asn
1175 1180 1185
His Cys Glu Tyr Asp Arg Gly Tyr Val Asn Tyr Pro Pro Val Pro
1190 1195 1200
Ala Gly Tyr Met Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp
1205 1210 1215
Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Lys Phe Ile Val
1220 1225 1230
Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1235 1240
<210> 42
<211> 3702
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_14.
<400> 42
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgaccgcgac gtttgaagct 1980
gaatccgacc tcgagcgtgc gcgcaaggcg gtgaacgctc tgttcacgag caccaaccct 2040
cgtggcttga agacggatgt gacggactac cacatcgacc aagtctcgaa cctcgtggag 2100
tgcctgagcg acgagttctg tcttgacaag aagcgcgagc tgctggagga ggtgaagtac 2160
gccaagcgcc tctccgatga gcgcaacctg ctccaagatc ctaccttcac gtcgatttcc 2220
ggccaaaccg accgtggatg gatcggctcg actggcatct ccatccaggg cggcgacgac 2280
atcttcaagg agaactatgt tcggctgccg ggcacggtgg acgagtgtta cccgacgtac 2340
ctctaccaga agatagacga gagtcaactc aagtcctaca cgcggtatca gttacgtggc 2400
tacattgaag actcccagga cttggaaatc tatctcatac ggtacaacgc caagcacgag 2460
accttaagcg tgccgggaac ggagtcgccc tggccaagct ctggcgtgta cccttccggt 2520
aggtgcggcg agcccaaccg ctgtgcacct cgaatcgaat ggaacccgga ccttgactgc 2580
tcttgccggt acggcgagaa gtgcgtccat cattctcacc acttcagctt ggacattgac 2640
gtcggctgca ccgacctcaa tgaagacctc ggagtgtggg tcatcttcaa gatcaagaca 2700
caggacgggc acgcgaaact aggaaacctg gagttcatcg aggagaagcc actcctcggc 2760
aaggcacttt ccagggtcaa gcgggccgag aagaaatgga gggacaagta cgagaaactc 2820
cagctcgaaa caaagcgggt gtacacggag gcaaaggaat ccgtggacgc cctgttcgtg 2880
gactctcagt acgacaagct ccaggcgaac acaaacattg gcatcatcca cggtgcggac 2940
aagcaagtgc acaggatacg ggagccttac ctctcggagc tgccggtgat tccctcgatc 3000
aacgcggcga tcttcgagga actggagggc cacatcttca aggcgtattc tctgtacgac 3060
gcgcgtaacg tcatcaagaa cggcgacttc aacaatgggc tgtcctgctg gaacgttaaa 3120
ggccacgtcg atgtccagca gaaccaccat aggtcagtcc tggtgctgag cgagtgggag 3180
gcggaggtgt cccagaaggt gcgcgtgtgc ccggatcgcg gctacatctt gagggtgaca 3240
gcctacaagg agggctacgg cgagggctgt gtcacgatcc atgagttcga ggacaacacg 3300
gatgtcctga aattccgtaa cttcgtcgag gaggaggtct atcccaacaa caccgtgacc 3360
tgcaacgact acacgaccaa tcagtcggct gagggcagta ccgatgcctg caacagctac 3420
aaccgtggtt acgaagatgg atacgagaac cgctacgagc ccaatccttc ggctcccgtg 3480
aattacactc ccacgtacga ggagggcatg tacactgaca ctcagggcta caaccattgc 3540
gtcagcgacc gtggctaccg caaccacacg ccgctcccag cgggctacgt gacgctggag 3600
ctggaatact ttcccgagac agaacaagtg tggatagaga tcggcgagac cgagggcaca 3660
ttcatcgtgg gctctgtgga attgctcctc atggaggagt aa 3702
<210> 43
<211> 1200
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_14.
<400> 43
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Thr Ala
645 650 655
Thr Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala Arg Lys Ala Val Asn
660 665 670
Ala Leu Phe Thr Ser Thr Asn Pro Arg Gly Leu Lys Thr Asp Val Thr
675 680 685
Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser Asp
690 695 700
Glu Phe Cys Leu Asp Lys Lys Arg Glu Leu Leu Glu Glu Val Lys Tyr
705 710 715 720
Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Thr Phe
725 730 735
Thr Ser Ile Ser Gly Gln Thr Asp Arg Gly Trp Ile Gly Ser Thr Gly
740 745 750
Ile Ser Ile Gln Gly Gly Asp Asp Ile Phe Lys Glu Asn Tyr Val Arg
755 760 765
Leu Pro Gly Thr Val Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys
770 775 780
Ile Asp Glu Ser Gln Leu Lys Ser Tyr Thr Arg Tyr Gln Leu Arg Gly
785 790 795 800
Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn
805 810 815
Ala Lys His Glu Thr Leu Ser Val Pro Gly Thr Glu Ser Pro Trp Pro
820 825 830
Ser Ser Gly Val Tyr Pro Ser Gly Arg Cys Gly Glu Pro Asn Arg Cys
835 840 845
Ala Pro Arg Ile Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Tyr
850 855 860
Gly Glu Lys Cys Val His His Ser His His Phe Ser Leu Asp Ile Asp
865 870 875 880
Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe
885 890 895
Lys Ile Lys Thr Gln Asp Gly His Ala Lys Leu Gly Asn Leu Glu Phe
900 905 910
Ile Glu Glu Lys Pro Leu Leu Gly Lys Ala Leu Ser Arg Val Lys Arg
915 920 925
Ala Glu Lys Lys Trp Arg Asp Lys Tyr Glu Lys Leu Gln Leu Glu Thr
930 935 940
Lys Arg Val Tyr Thr Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val
945 950 955 960
Asp Ser Gln Tyr Asp Lys Leu Gln Ala Asn Thr Asn Ile Gly Ile Ile
965 970 975
His Gly Ala Asp Lys Gln Val His Arg Ile Arg Glu Pro Tyr Leu Ser
980 985 990
Glu Leu Pro Val Ile Pro Ser Ile Asn Ala Ala Ile Phe Glu Glu Leu
995 1000 1005
Glu Gly His Ile Phe Lys Ala Tyr Ser Leu Tyr Asp Ala Arg Asn
1010 1015 1020
Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn
1025 1030 1035
Val Lys Gly His Val Asp Val Gln Gln Asn His His Arg Ser Val
1040 1045 1050
Leu Val Leu Ser Glu Trp Glu Ala Glu Val Ser Gln Lys Val Arg
1055 1060 1065
Val Cys Pro Asp Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr Lys
1070 1075 1080
Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Phe Glu Asp
1085 1090 1095
Asn Thr Asp Val Leu Lys Phe Arg Asn Phe Val Glu Glu Glu Val
1100 1105 1110
Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp Tyr Thr Thr Asn Gln
1115 1120 1125
Ser Ala Glu Gly Ser Thr Asp Ala Cys Asn Ser Tyr Asn Arg Gly
1130 1135 1140
Tyr Glu Asp Gly Tyr Glu Asn Arg Tyr Glu Pro Asn Pro Ser Ala
1145 1150 1155
Pro Val Asn Tyr Thr Pro Thr Tyr Glu Glu Gly Met Tyr Thr Asp
1160 1165 1170
Thr Gln Gly Tyr Asn His Cys Val Ser Asp Arg Gly Tyr Arg Asn
1175 1180 1185
His Thr Pro Leu Pro Ala Gly Tyr Val Thr Leu Glu
1190 1195 1200
<210> 44
<211> 3687
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_15.
<400> 44
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagcagtc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctccgtgc tctcacgctg gtccaacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cggatgctac ctttgaagca 1980
gagtccgact tggaacgtgc acagaaggca gtgaacgcac tcttcacctc aagcaaccag 2040
atcggattga agacagatgt gacagattac cacatcgacc aagtgagcaa cttggtggat 2100
tgcttgtcag atgagttctg cttggatgag aagcgtgaac tctccgagaa ggtgaagcac 2160
gcaaagcgtc tctcagatga acgtaatctc cttcaagacc ctaactttcg tggtatcaat 2220
cgtcagccag atcgtggatg gcgtggatca acagacatca ccatccaggg aggcgatgat 2280
gtgttcaagg agaactacgt gaccctccca ggaaccgtgg atgaatgcta cccaacctac 2340
ctctaccaga agatcgacga gtcaaagctc aaggcttaca cccgttatga actccgtggc 2400
tacatcgaag atagccagga tctcgaaatc tatctcatcc gttacaatgc taagcacgaa 2460
atcgtgaatg tgccaggaac cggctcactc tggccactct cagcacagtc accaatcggc 2520
aagtgcggcg aacccaatcg ctgcgctcct catctcgaat ggaatcccga tctcgactgc 2580
tcctgccgag acggcgagaa gtgtgcacat cactcacacc acttcaccct cgacatcgac 2640
gtgggctgca ccgacctcaa tgaagacctg ggcgtgtggg tgatcttcaa gatcaagacc 2700
caggacggcc acgcacgact gggcaatctg gagtttctgg aggagaagcc actgcttggc 2760
gaggcactgg cacgagtgaa acgagccgag aagaaatggc gagacaaacg tgagaagctg 2820
caactggaga ccaacatcgt gtacaaagag gccaaagagt cagttgacgc cctgtttgtc 2880
aatagccagt atgaccgact gcaagttgac accaacatcg ccatgatcca cgctgcggac 2940
aagcgcgtcc accgcatccg cgaggcttat ctgcccgagc tgagcgtcat tcccggcgtc 3000
aatgccgcga tcttcgagga gttagagggc cgcatcttca ccgcctacag cctctatgac 3060
gcccgcaatg tcattaagaa tggcgacttc aacaatggct tactatgctg gaatgtcaaa 3120
gggcacgttg acgtcgagga gcagaacaat caccgcagcg tcttagtcat acccgagtgg 3180
gaggccgaag tcagccagga agtccgcgtc tgtccagggc gcgggtacat cctgcgggtc 3240
accgcctaca aagagggata cggcgagggt tgtgtcacca tacacgagat agaggacaat 3300
accgacgaac tcaagttcag caattgtgtc gaggaggaag tctatcccaa caataccgta 3360
acctgcaaca actacaccgg aacccaggag gagtatgaag ggacgtacac ctcgcggaac 3420
cagggctatg acgaagccta tgggaacaac ccgtcggtgc ctgctgacta tgcgtcggtc 3480
tatgaggaga aatcgtacac ggacgggcgg cgggagaatc cgtgtgagtc gaatcgcggg 3540
tatggtgact acacgccgct accggcgggc tatgtaacga aagacctgga atacttcccg 3600
gagacggaca aagtatggat agagataggc gagacggagg gaacgttcat cgtggactcg 3660
gtagagctgc tgctcatgga ggagtga 3687
<210> 45
<211> 1228
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_15.
<400> 45
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Gln Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Val Leu Ser Arg Trp Ser Asn Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Asp Ala
645 650 655
Thr Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn
660 665 670
Ala Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr
675 680 685
Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp
690 695 700
Glu Phe Cys Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys His
705 710 715 720
Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe
725 730 735
Arg Gly Ile Asn Arg Gln Pro Asp Arg Gly Trp Arg Gly Ser Thr Asp
740 745 750
Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr
755 760 765
Leu Pro Gly Thr Val Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys
770 775 780
Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Glu Leu Arg Gly
785 790 795 800
Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn
805 810 815
Ala Lys His Glu Ile Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro
820 825 830
Leu Ser Ala Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys
835 840 845
Ala Pro His Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp
850 855 860
Gly Glu Lys Cys Ala His His Ser His His Phe Thr Leu Asp Ile Asp
865 870 875 880
Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe
885 890 895
Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe
900 905 910
Leu Glu Glu Lys Pro Leu Leu Gly Glu Ala Leu Ala Arg Val Lys Arg
915 920 925
Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Gln Leu Glu Thr
930 935 940
Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val
945 950 955 960
Asn Ser Gln Tyr Asp Arg Leu Gln Val Asp Thr Asn Ile Ala Met Ile
965 970 975
His Ala Ala Asp Lys Arg Val His Arg Ile Arg Glu Ala Tyr Leu Pro
980 985 990
Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu
995 1000 1005
Glu Gly Arg Ile Phe Thr Ala Tyr Ser Leu Tyr Asp Ala Arg Asn
1010 1015 1020
Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Leu Cys Trp Asn
1025 1030 1035
Val Lys Gly His Val Asp Val Glu Glu Gln Asn Asn His Arg Ser
1040 1045 1050
Val Leu Val Ile Pro Glu Trp Glu Ala Glu Val Ser Gln Glu Val
1055 1060 1065
Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala Tyr
1070 1075 1080
Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile Glu
1085 1090 1095
Asp Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val Glu Glu Glu
1100 1105 1110
Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asn Tyr Thr Gly Thr
1115 1120 1125
Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Gln Gly Tyr
1130 1135 1140
Asp Glu Ala Tyr Gly Asn Asn Pro Ser Val Pro Ala Asp Tyr Ala
1145 1150 1155
Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg Glu Asn
1160 1165 1170
Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu Pro
1175 1180 1185
Ala Gly Tyr Val Thr Lys Asp Leu Glu Tyr Phe Pro Glu Thr Asp
1190 1195 1200
Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val
1205 1210 1215
Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1220 1225
<210> 46
<211> 3600
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC868_29.
<400> 46
atgacgagca accggaagaa cgagaacgag atcatcaacg ccctctcgat ccctgctgtt 60
tcaaaccact ccgcgcagat gaacctgtcc accgacgcgc gcatcgagga ctccctctgc 120
atagccgagg gcaacaacat cgacccattc gtgtcggcca gcacggttca gaccggcatc 180
aacatcgcgg gccgtatcct cggcgtcctc ggtgtcccat tcgccggtca gatcgcgtcc 240
ttctactcgt tccttgtggg cgagctgtgg cctcgcggtc gtgacccgtg ggagatcttc 300
ctggagcatg tggagcagtt gatccggcag caagtcacgg agaacacccg cgatactgct 360
ctggccaggc tacagggcct gggaaactcc tttcgggcat accagtactc actggaggac 420
tggttggaga acagggatga cgcgcgaaca cgctcggtac tctacaccca gtacatcgct 480
ctcgaactcg acttcctgaa cgctatgccg ctgttcgcca tcaggaacca ggaagttcca 540
ctccttatgg tgtacgccca ggccgccaac ttacatctgc tcctgctgcg ggacgccagc 600
ctgttcggct ccgagttcgg actcacatct caagaaatcc agcgttacta cgagcgccaa 660
gtggagaaga cccgtgagta cagtgactac tgcgctcgat ggtacaacac agggctcaac 720
aacctgcgcg gcaccaacgc tgagtcatgg ctccgttaca accagttccg ccgcgacttg 780
actttgggtg tcctagacct ggtggcgcta ttcccgtctt acgacacacg ggtgtaccca 840
atgaacacta gcgcgcaact cacgcgggag atctacacag acccaatcgg ccggacgaac 900
gcaccctccg gtttcgcatc cacgaattgg ttcaacaaca acgcaccctc cttctcggca 960
atcgaggccg ccgtcatccg ccctcctcac ctgctcgact ttcccgagca gctcacgatc 1020
ttctcccagc tctcacgctg gtcccacaca cagtacatga actactgggt cgggcaccga 1080
ttggagagta ggacgatccg tggcagcttg agcaccagta cccacggcaa caccaacacc 1140
tccatcaacc cagttacgct acagttcacg agccgcgacg tttaccggac tgagtcgttc 1200
gcgggcatta acatccttct gacaacgccc gtcaacggcg tcccgtgggc ccggttcaac 1260
tggcgtaacc cgttgaactc cctgcgcggg tcattgctct acaccatcgg gtacacgggc 1320
gtcggcaccc agctcttcga cagtgaaact gagctgccgc ccgagaccac ggaacgcccg 1380
aactacgagt cctacagcca ccgcctgtcc aacatccggc tcatctctgg caacacgctg 1440
cgtgcgccgg tgtactcctg gacacaccgc agcgccgacc ggaccaacac gatctcttcc 1500
gactccatta accagatccc gctcgtgaag ggcttccgtg tgtggggtgg cacgagcgtc 1560
atcaccggtc cgggcttcac cggtggagac atactgcggc gcaacacttt cggcgacttc 1620
gtttcgttgc aagtgaacat caactcgccg atcacccagc gttaccgtct gaggttccgc 1680
tacgcttcaa gccgcgacgc gagggtcatt gtcctgaccg gagccgcgtc cacaggcgtg 1740
ggaggccaag tctcagtcaa catgcctctc cagaagacga tggagatagg cgagaacttg 1800
actagccgaa ccttccggta cactgatttc tcgaaccctt tctcattcag agcgaaccct 1860
gacatcattg ggatctccga gcaaccgctg ttcggtgctg gctccatcag ctctggcgaa 1920
ctgtacatcg acaagattga gatcatcctg gcggatgcga cgttcgaggc cgagtctgac 1980
ctggagcggg ctcagaaggc tgtcaacgaa ctgttcacca gcagcaacca gattgggctc 2040
aagaccgacg tcacggacta tcacattgac caagtgtcca accttgtgga gtgcctgtcc 2100
gacgagttct gcctcgacga gaagaaggag ctgtccgaga aggtcaaaca cgcgaagcgt 2160
ctgagtgacg agcggaattt gctccaggac ccgaacttcc gtggcatcaa ccgccagctc 2220
gaccgtggtt ggcgcgggag tacagacatc accatccagg gaggcgacga tgtgttcaag 2280
gagaactatg tgacgctgct cgggactttc gacgaatgct acccgacgta tctctaccag 2340
aagatagacg agagtaaatt gaaggcgtac acccgctacc agcttcgcgg gtacatcgag 2400
gatagtcagg acctggaaat ctacctgatc cgatacaacg ccaagcacga gacagtgaac 2460
gtgccaggca cgggctcact ttggccattg agcgctccct ctccaatcgg aaagtgcgct 2520
caccactcgc accacttctc tctggacatc gacgtgggct gcaccgacct caacgaggac 2580
ctgggtgtct gggttatctt caagattaag acccaggacg gacatgcccg cctcggcaac 2640
ctggagttcc ttgaggagaa gcctctcgtg ggcgaggccc tcgctcgtgt gaagcgcgcc 2700
gagaagaaat ggcgagacaa gcgggagaag ctggagtggg agaccaacat cgtgtacaag 2760
gaggccaagg agtcagtgga cgcactcttc gtcaacagcc agtacgaccg cctccaggct 2820
gacaccaaca tcgccatgat ccacgcggct gacaagcggg tccacagcat ccgtgaggcg 2880
tacctgcccg agctgtcagt gatccctggt gtgaacgcgg cgatcttcga ggaactggag 2940
ggccgcatct tcacagcatt cagcctgtac gatgccagga atgttattaa gaacggtgac 3000
ttcaacaacg ggctgagttg ctggaacgtc aagggccatg tggacgtcga ggagcagaac 3060
aaccaccggt ccgtgctggt cgtgccggag tgggaggcag aggtgagcca ggaggtccgc 3120
gtctgccctg gtcgcggcta catcctccgt gtgactgcgt acaaggaagg ctacggtgaa 3180
ggctgcgtga ctatccacga gatcgagaac aacaccgacg agctcaagtt ctcgaactgt 3240
gtggaggagg aggtgtaccc gaacaacacc gttacttgca acgactacac tgccacgcaa 3300
gaggagtacg agggcactta cacttcccgg aatcgcggct atgatggcgc gtacgagtcc 3360
aacagcagcg tgcctgcgga ttatgcgtcc gcttacgagg agaaggcgta caccgacgga 3420
cggagggaca acccttgcga gtccaaccgt ggctacggtg actacactcc gctgcccgcc 3480
gggtacgtca ccaaggagct ggagtacttc ccggagaccg acaaagtctg gatcgagatc 3540
ggcgagacgg agggcacttt catcgtggac tcggtcgagc tgctactgat ggaggagtga 3600
<210> 47
<211> 1199
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein variant TIC868_29.
<400> 47
Met Thr Ser Asn Arg Lys Asn Glu Asn Glu Ile Ile Asn Ala Leu Ser
1 5 10 15
Ile Pro Ala Val Ser Asn His Ser Ala Gln Met Asn Leu Ser Thr Asp
20 25 30
Ala Arg Ile Glu Asp Ser Leu Cys Ile Ala Glu Gly Asn Asn Ile Asp
35 40 45
Pro Phe Val Ser Ala Ser Thr Val Gln Thr Gly Ile Asn Ile Ala Gly
50 55 60
Arg Ile Leu Gly Val Leu Gly Val Pro Phe Ala Gly Gln Ile Ala Ser
65 70 75 80
Phe Tyr Ser Phe Leu Val Gly Glu Leu Trp Pro Arg Gly Arg Asp Pro
85 90 95
Trp Glu Ile Phe Leu Glu His Val Glu Gln Leu Ile Arg Gln Gln Val
100 105 110
Thr Glu Asn Thr Arg Asp Thr Ala Leu Ala Arg Leu Gln Gly Leu Gly
115 120 125
Asn Ser Phe Arg Ala Tyr Gln Tyr Ser Leu Glu Asp Trp Leu Glu Asn
130 135 140
Arg Asp Asp Ala Arg Thr Arg Ser Val Leu Tyr Thr Gln Tyr Ile Ala
145 150 155 160
Leu Glu Leu Asp Phe Leu Asn Ala Met Pro Leu Phe Ala Ile Arg Asn
165 170 175
Gln Glu Val Pro Leu Leu Met Val Tyr Ala Gln Ala Ala Asn Leu His
180 185 190
Leu Leu Leu Leu Arg Asp Ala Ser Leu Phe Gly Ser Glu Phe Gly Leu
195 200 205
Thr Ser Gln Glu Ile Gln Arg Tyr Tyr Glu Arg Gln Val Glu Lys Thr
210 215 220
Arg Glu Tyr Ser Asp Tyr Cys Ala Arg Trp Tyr Asn Thr Gly Leu Asn
225 230 235 240
Asn Leu Arg Gly Thr Asn Ala Glu Ser Trp Leu Arg Tyr Asn Gln Phe
245 250 255
Arg Arg Asp Leu Thr Leu Gly Val Leu Asp Leu Val Ala Leu Phe Pro
260 265 270
Ser Tyr Asp Thr Arg Val Tyr Pro Met Asn Thr Ser Ala Gln Leu Thr
275 280 285
Arg Glu Ile Tyr Thr Asp Pro Ile Gly Arg Thr Asn Ala Pro Ser Gly
290 295 300
Phe Ala Ser Thr Asn Trp Phe Asn Asn Asn Ala Pro Ser Phe Ser Ala
305 310 315 320
Ile Glu Ala Ala Val Ile Arg Pro Pro His Leu Leu Asp Phe Pro Glu
325 330 335
Gln Leu Thr Ile Phe Ser Gln Leu Ser Arg Trp Ser His Thr Gln Tyr
340 345 350
Met Asn Tyr Trp Val Gly His Arg Leu Glu Ser Arg Thr Ile Arg Gly
355 360 365
Ser Leu Ser Thr Ser Thr His Gly Asn Thr Asn Thr Ser Ile Asn Pro
370 375 380
Val Thr Leu Gln Phe Thr Ser Arg Asp Val Tyr Arg Thr Glu Ser Phe
385 390 395 400
Ala Gly Ile Asn Ile Leu Leu Thr Thr Pro Val Asn Gly Val Pro Trp
405 410 415
Ala Arg Phe Asn Trp Arg Asn Pro Leu Asn Ser Leu Arg Gly Ser Leu
420 425 430
Leu Tyr Thr Ile Gly Tyr Thr Gly Val Gly Thr Gln Leu Phe Asp Ser
435 440 445
Glu Thr Glu Leu Pro Pro Glu Thr Thr Glu Arg Pro Asn Tyr Glu Ser
450 455 460
Tyr Ser His Arg Leu Ser Asn Ile Arg Leu Ile Ser Gly Asn Thr Leu
465 470 475 480
Arg Ala Pro Val Tyr Ser Trp Thr His Arg Ser Ala Asp Arg Thr Asn
485 490 495
Thr Ile Ser Ser Asp Ser Ile Asn Gln Ile Pro Leu Val Lys Gly Phe
500 505 510
Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly
515 520 525
Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln
530 535 540
Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg
545 550 555 560
Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala
565 570 575
Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys
580 585 590
Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr
595 600 605
Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly
610 615 620
Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu
625 630 635 640
Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu
645 650 655
Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Glu Leu Phe
660 665 670
Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His
675 680 685
Ile Asp Gln Val Ser Asn Leu Val Glu Cys Leu Ser Asp Glu Phe Cys
690 695 700
Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg
705 710 715 720
Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile
725 730 735
Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile
740 745 750
Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Leu Gly
755 760 765
Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu
770 775 780
Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu
785 790 795 800
Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His
805 810 815
Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala
820 825 830
Pro Ser Pro Ile Gly Lys Cys Ala His His Ser His His Phe Ser Leu
835 840 845
Asp Ile Asp Val Gly Cys Thr Asp Leu Asn Glu Asp Leu Gly Val Trp
850 855 860
Val Ile Phe Lys Ile Lys Thr Gln Asp Gly His Ala Arg Leu Gly Asn
865 870 875 880
Leu Glu Phe Leu Glu Glu Lys Pro Leu Val Gly Glu Ala Leu Ala Arg
885 890 895
Val Lys Arg Ala Glu Lys Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu
900 905 910
Trp Glu Thr Asn Ile Val Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala
915 920 925
Leu Phe Val Asn Ser Gln Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile
930 935 940
Ala Met Ile His Ala Ala Asp Lys Arg Val His Ser Ile Arg Glu Ala
945 950 955 960
Tyr Leu Pro Glu Leu Ser Val Ile Pro Gly Val Asn Ala Ala Ile Phe
965 970 975
Glu Glu Leu Glu Gly Arg Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala
980 985 990
Arg Asn Val Ile Lys Asn Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp
995 1000 1005
Asn Val Lys Gly His Val Asp Val Glu Glu Gln Asn Asn His Arg
1010 1015 1020
Ser Val Leu Val Val Pro Glu Trp Glu Ala Glu Val Ser Gln Glu
1025 1030 1035
Val Arg Val Cys Pro Gly Arg Gly Tyr Ile Leu Arg Val Thr Ala
1040 1045 1050
Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr Ile His Glu Ile
1055 1060 1065
Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser Asn Cys Val Glu Glu
1070 1075 1080
Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn Asp Tyr Thr Ala
1085 1090 1095
Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg Asn Arg Gly
1100 1105 1110
Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala Asp Tyr
1115 1120 1125
Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg Asp
1130 1135 1140
Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu
1145 1150 1155
Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr
1160 1165 1170
Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile
1175 1180 1185
Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1190 1195
<210> 48
<211> 3432
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC869.
<400> 48
atggagataa ataatcagaa gcaatgcata ccatataatt gcttaagtaa tcctgaggaa 60
gtacttttgg atggggagag gatattacct gatatcgatc cactcgaagt ttctttgtcg 120
cttttgcaat ttcttttgaa taactttgtt ccagggggag gctttatttc aggattagtt 180
gataaaatat ggggggcttt gagaccatct gaatgggact tatttcttgc acagattgaa 240
cggttgattg atcaaagaat agaagcaaca gtaagagcaa aagcaatcac tgaattagaa 300
ggattaggga gaaattatca aatatacgct gaagcattta aagaatggga atcagatcct 360
gataacgaag cggctaaaag tagagtaatt gatcgctttc gtatacttga tggtctaatt 420
gaagcaaata tcccttcatt tcggataatt ggatttgaag tgccactttt atcggtttat 480
gttcaagcag ctaatctaca tctcgctcta ttgagagatt ctgttatttt tggagagaga 540
tggggattga cgacaaaaaa tgtcaatgat atctataata gacaaattag agaaattcat 600
gaatatagca atcattgcgt agatacgtat aacacagaac tagaacgtct agggtttaga 660
tctatagcgc agtggagaat atataatcag tttagaagag aactaacact aactgtatta 720
gatattgtcg ctcttttccc gaactatgac agtagactgt atccgatcca aactttttct 780
caattgacaa gagaaattgt tacatcccca gtaagcgaat tttattatgg tgttattaat 840
agtggtaata taattggtac tcttactgaa cagcagataa ggcgaccaca tcttatggac 900
ttctttaact ccatgatcat gtatacatca gataatagac gggaacatta ttggtcagga 960
cttgaaatga cggcttattt tacaggattt gcaggagctc aagtgtcatt ccctttagtc 1020
gggactagag gggagtcagc tccaccatta actgttagaa gtgttaatga tggaatttat 1080
agaatattat cggcaccgtt ttattcagcg ccttttctag gcaccattgt attgggaagt 1140
cgtggagaaa aatttgattt tgcgcttaat aatatttcac ctccgccatc tacaatatac 1200
agacatcctg gaacagtaga ttcactagtc agtataccgc cacaggataa tagcgtacca 1260
ccgcacaggg gatctagtca tcgattaagt catgttacaa tgcgcgcaag ttcccctata 1320
ttccattgga cgcatcgcag cgcaaccact acaaatacaa ttaatccaaa tgctattatc 1380
caaataccac tagtaaaagc atttaacctt cattcaggtg ccactgttgt tagaggacca 1440
gggtttacag gtggagatct cttacgaaga acgaatactg gtacatttgc agacataaga 1500
gtcaatgttc cttcatcact attttcccaa agatatcgcg taaggattcg ttatgcttct 1560
actaccgatt tacaattttt cacgagaatt aatggaactt ctgttaatca aggtaatttc 1620
tcaaaaacga tggatagagg ggataaactg aaatctgaaa actttagaac tgccggattt 1680
agtactcctt ttagattttc aaattttcaa agtacattca cgttgggtac tcaggctttt 1740
tcaaatcagg aagtttatat agatagaatt gaatttgtcc cggcagaagt aacattcgag 1800
gcagaatctg atttagaaag agcacaaaag gcggtgaatg agctgtttac ttcttccaat 1860
caaatcgggt taaaaacaga tgtgacggat tatcatattg atcaagtatc caatttagtt 1920
gagtgtttat ctgatgaatt ttgtctggat gaaaaaaaag aattgtccga gaaagtcaaa 1980
catgcgaagc gacttagtga tgagcggaat ttacttcaag atccaaactt tagagggatc 2040
aatagacaac tagaccgtgg ctggagagga agtacggata ttaccatcca aggaggcgat 2100
gacgtattca aagagaatta cgttacgcta ttgggtacct ttgatgagtg ctatccaacg 2160
tatttatatc aaaaaataga tgagtcgaaa ttaaaagcct atacccgtta ccaattaaga 2220
gggtatatcg aagatagtca agacttagaa atctatttaa ttcgctacaa tgccaaacac 2280
gaaacagtaa atgtgccagg tacgggttcc ttatggccgc tttcagcccc aagtccaatc 2340
ggaaaatgtg cccatcattc ccatcatttc tccttggaca ttgatgttgg atgtacagac 2400
ttaaatgagg acttaggtgt atgggtgata ttcaagatta agacgcaaga tggccatgca 2460
agactaggaa atctagaatt tctcgaagag aaaccattag taggagaagc actagctcgt 2520
gtgaaaagag cggagaaaaa atggagagac aaacgtgaaa aattggaatg ggaaacaaat 2580
attgtttata aagaggcaaa agaatctgta gatgctttat ttgtaaactc tcaatatgat 2640
agattacaag cggataccaa catcgcgatg attcatgcgg cagataaacg cgttcatagc 2700
attcgagaag cttatctgcc tgagctgtct gtgattccgg gtgtcaatgc ggctattttt 2760
gaagaattag aagggcgtat tttcactgca ttctccctat atgatgcgag aaatgtcatt 2820
aaaaatggtg attttaataa tggcttatcc tgctggaacg tgaaagggca tgtagatgta 2880
gaagaacaaa acaaccaccg ttcggtcctt gttgttccgg aatgggaagc agaagtgtca 2940
caagaagttc gtgtctgtcc gggtcgtggc tatatccttc gtgtcacagc gtacaaggag 3000
ggatatggag aaggttgcgt aaccattcat gagatcgaga acaatacaga cgaactgaag 3060
tttagcaact gtgtagaaga ggaagtatat ccaaacaaca cggtaacgtg taatgattat 3120
actgcgactc aagaagaata tgagggtacg tacacttctc gtaatcgagg atatgacgga 3180
gcctatgaaa gcaattcttc tgtaccagct gattatgcat cagcctatga agaaaaagca 3240
tatacagatg gacgaagaga caatccttgt gaatctaaca gaggatatgg ggattacaca 3300
ccactaccag ctggctatgt gacaaaagaa ttagagtact tcccagaaac cgataaggta 3360
tggattgaga tcggagaaac ggaaggaaca ttcatcgtgg acagcgtgga attacttctt 3420
atggaggaat ag 3432
<210> 49
<211> 3432
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC869.
<400> 49
atggagataa acaaccagaa gcagtgcatt ccgtacaact gcctcagcaa cccggaggag 60
gtgctgctgg acggcgagcg tatcctccca gacatcgacc cactggaggt cagcctgagc 120
ctcctccagt tcctcctcaa taacttcgtg ccaggcggcg gcttcatctc cggcctggtg 180
gacaagatct ggggcgcact ccggccaagt gagtgggatc tgttcctggc ccaaatcgag 240
cgcctgatcg accagaggat cgaggcgacg gtccgcgcca aggcgataac cgagctggag 300
ggcctcggtc gcaactacca gatctacgca gaggcgttca aggagtggga gagcgacccg 360
gacaacgagg cggccaagtc tcgggtgatt gaccgcttcc gcatcctcga cggcctcatc 420
gaagccaaca tcccttcctt ccggatcata ggcttcgaag tcccgctcct cagcgtgtac 480
gtgcaagcgg ccaatctcca cctcgcgttg ctccgtgaca gcgtcatctt tggcgagaga 540
tggggcctga cgacgaagaa cgtgaacgac atctacaaca ggcagatccg agagattcac 600
gagtacagca accactgcgt ggacacatac aacacggagc tggagcggct cggcttccgc 660
tcaatcgctc agtggcggat ctacaaccag ttccgccgcg agctgaccct caccgtgctc 720
gacatcgtcg cattgtttcc caattacgac tcacgcctct acccaatcca gactttcagc 780
cagctcacac gcgagattgt gaccagcccg gtgtcagagt tctactacgg cgtcatcaac 840
tcaggcaaca tcatcgggac actgactgaa cagcagatca gacgtccgca cttgatggac 900
ttcttcaact ccatgattat gtacacatca gacaacagga gagagcacta ctggtccggg 960
ttggagatga ctgcttactt caccggcttc gccggtgccc aagtgagctt cccactggtc 1020
ggaactcgtg gcgagtcagc tcctccgcta actgtgcgat ctgtcaacga cgggatctac 1080
agaatactgt cggctccctt ctacagtgcg ccgttcctcg gcaccatcgt cctcggctca 1140
cgtggtgaga agttcgactt cgcactgaac aacattagcc cgccgcctag tacaatctac 1200
aggcaccctg gcaccgtgga ctcactggtt tcgatcccgc cacaagacaa cagtgtgccg 1260
ccacatcgtg gttctagcca caggctctcc catgtgacca tgcgcgcctc ttcaccgatc 1320
tttcactgga cccatcggtc cgctacaacc acaaacacca tcaaccctaa cgccatcatc 1380
caaatcccgc tggtgaaggc gtttaacctc cacagcggcg caactgtcgt gcgcggccct 1440
ggattcaccg gtggtgacct gctccgtcgg accaatactg gcacgttcgc agacatccga 1500
gtgaacgtcc cgtcctcgct gttcagtcag cgctaccgtg tccgcattcg gtacgcttcc 1560
accacggatc tccagttctt tactcgcatc aatgggacga gcgtcaacca gggcaacttc 1620
agcaagacga tggaccgtgg agataagctc aagtccgaga acttccgcac ggctggcttc 1680
tcgacaccgt tcagattcag caacttccag agcactttca cgctgggcac acaggcgttc 1740
tccaaccagg aggtgtacat cgaccgcatc gagttcgtgc ctgctgaggt taccttcgag 1800
gcggaaagcg acctcgaaag ggcccagaag gccgtcaacg agctgttcac ctccagcaac 1860
cagatcggtc tcaagaccga cgtcactgac tatcacattg accaagtcag caacctggtg 1920
gagtgcctca gtgatgagtt ctgcctggat gagaagaagg agcttagcga gaaggtcaag 1980
cacgcaaagc gcttgagcga cgagcgcaac cttctccagg acccgaattt ccgtggtatc 2040
aatagacagc ttgaccgtgg gtggcgcggt agtaccgaca taaccatcca gggtggcgac 2100
gatgtgttca aggagaatta tgttacgctg ctcggtacgt tcgacgagtg ctatcccacg 2160
tacttgtacc agaagattga cgagagcaag ctcaaggcgt acacccgtta ccagctccgt 2220
ggctacatcg aggacagcca ggatctggaa atctacctta tccgatacaa tgctaagcac 2280
gagacagtca acgtgcccgg aacagggtcg ctctggccgc tcagtgctcc gtcgcccatt 2340
ggcaagtgcg cgcaccattc gcatcacttc tcacttgaca ttgacgtggg ctgcaccgac 2400
ctgaacgagg atctgggtgt ctgggtcatc ttcaagatca agacccaaga cggccacgcg 2460
cgcctcggga acctggagtt cctggaggag aagcctttgg taggtgaagc cctggcccgc 2520
gtcaagcgcg cggagaagaa gtggcgcgac aagagggaga agctggaatg ggagaccaac 2580
atcgtgtaca aggaggcgaa ggagtcggtg gacgcactat tcgtgaactc ccagtacgac 2640
cgtctccagg ccgacaccaa catcgccatg atccacgccg ctgacaaacg agttcattcc 2700
attcgtgaag cctatcttcc cgagctgtct gtcataccgg gcgtcaacgc ggccatcttc 2760
gaggagttag agggtcggat ctttacagct ttctcactgt acgatgcccg caacgtcatc 2820
aagaacggcg acttcaacaa cggtctctcc tgttggaacg tgaagggcca cgtggatgtc 2880
gaggagcaga acaaccaccg ctctgtgctt gtggtgcccg agtgggaggc cgaggtgagc 2940
caggaggtcc gcgtctgtcc gggtcgcggc tacatcctgc gggtcaccgc ctacaaggag 3000
ggctacggcg aaggctgcgt tactattcac gagattgaga acaataccga cgaactcaag 3060
ttctccaact gtgtcgagga ggaggtgtac ccgaacaaca ccgtgacgtg caacgactac 3120
accgcgacac aggaggaata cgagggcacc tacaccagcc gcaaccgagg ctacgacgga 3180
gcgtacgaga gcaactcgtc cgtgcccgct gattacgcga gtgcgtacga ggagaaggct 3240
tacaccgacg gacggcgcga caatccctgc gagagtaacc gtggatacgg agattacacg 3300
ccgctacccg ctggctacgt cactaaggaa ctggagtact tcccagagac ggacaaggtg 3360
tggatcgaaa tcggcgagac agagggcacg ttcatcgtgg actccgtgga gctgctgctg 3420
atggaggagt ga 3432
<210> 50
<211> 1143
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein TIC869.
<400> 50
Met Glu Ile Asn Asn Gln Lys Gln Cys Ile Pro Tyr Asn Cys Leu Ser
1 5 10 15
Asn Pro Glu Glu Val Leu Leu Asp Gly Glu Arg Ile Leu Pro Asp Ile
20 25 30
Asp Pro Leu Glu Val Ser Leu Ser Leu Leu Gln Phe Leu Leu Asn Asn
35 40 45
Phe Val Pro Gly Gly Gly Phe Ile Ser Gly Leu Val Asp Lys Ile Trp
50 55 60
Gly Ala Leu Arg Pro Ser Glu Trp Asp Leu Phe Leu Ala Gln Ile Glu
65 70 75 80
Arg Leu Ile Asp Gln Arg Ile Glu Ala Thr Val Arg Ala Lys Ala Ile
85 90 95
Thr Glu Leu Glu Gly Leu Gly Arg Asn Tyr Gln Ile Tyr Ala Glu Ala
100 105 110
Phe Lys Glu Trp Glu Ser Asp Pro Asp Asn Glu Ala Ala Lys Ser Arg
115 120 125
Val Ile Asp Arg Phe Arg Ile Leu Asp Gly Leu Ile Glu Ala Asn Ile
130 135 140
Pro Ser Phe Arg Ile Ile Gly Phe Glu Val Pro Leu Leu Ser Val Tyr
145 150 155 160
Val Gln Ala Ala Asn Leu His Leu Ala Leu Leu Arg Asp Ser Val Ile
165 170 175
Phe Gly Glu Arg Trp Gly Leu Thr Thr Lys Asn Val Asn Asp Ile Tyr
180 185 190
Asn Arg Gln Ile Arg Glu Ile His Glu Tyr Ser Asn His Cys Val Asp
195 200 205
Thr Tyr Asn Thr Glu Leu Glu Arg Leu Gly Phe Arg Ser Ile Ala Gln
210 215 220
Trp Arg Ile Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val Leu
225 230 235 240
Asp Ile Val Ala Leu Phe Pro Asn Tyr Asp Ser Arg Leu Tyr Pro Ile
245 250 255
Gln Thr Phe Ser Gln Leu Thr Arg Glu Ile Val Thr Ser Pro Val Ser
260 265 270
Glu Phe Tyr Tyr Gly Val Ile Asn Ser Gly Asn Ile Ile Gly Thr Leu
275 280 285
Thr Glu Gln Gln Ile Arg Arg Pro His Leu Met Asp Phe Phe Asn Ser
290 295 300
Met Ile Met Tyr Thr Ser Asp Asn Arg Arg Glu His Tyr Trp Ser Gly
305 310 315 320
Leu Glu Met Thr Ala Tyr Phe Thr Gly Phe Ala Gly Ala Gln Val Ser
325 330 335
Phe Pro Leu Val Gly Thr Arg Gly Glu Ser Ala Pro Pro Leu Thr Val
340 345 350
Arg Ser Val Asn Asp Gly Ile Tyr Arg Ile Leu Ser Ala Pro Phe Tyr
355 360 365
Ser Ala Pro Phe Leu Gly Thr Ile Val Leu Gly Ser Arg Gly Glu Lys
370 375 380
Phe Asp Phe Ala Leu Asn Asn Ile Ser Pro Pro Pro Ser Thr Ile Tyr
385 390 395 400
Arg His Pro Gly Thr Val Asp Ser Leu Val Ser Ile Pro Pro Gln Asp
405 410 415
Asn Ser Val Pro Pro His Arg Gly Ser Ser His Arg Leu Ser His Val
420 425 430
Thr Met Arg Ala Ser Ser Pro Ile Phe His Trp Thr His Arg Ser Ala
435 440 445
Thr Thr Thr Asn Thr Ile Asn Pro Asn Ala Ile Ile Gln Ile Pro Leu
450 455 460
Val Lys Ala Phe Asn Leu His Ser Gly Ala Thr Val Val Arg Gly Pro
465 470 475 480
Gly Phe Thr Gly Gly Asp Leu Leu Arg Arg Thr Asn Thr Gly Thr Phe
485 490 495
Ala Asp Ile Arg Val Asn Val Pro Ser Ser Leu Phe Ser Gln Arg Tyr
500 505 510
Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asp Leu Gln Phe Phe Thr
515 520 525
Arg Ile Asn Gly Thr Ser Val Asn Gln Gly Asn Phe Ser Lys Thr Met
530 535 540
Asp Arg Gly Asp Lys Leu Lys Ser Glu Asn Phe Arg Thr Ala Gly Phe
545 550 555 560
Ser Thr Pro Phe Arg Phe Ser Asn Phe Gln Ser Thr Phe Thr Leu Gly
565 570 575
Thr Gln Ala Phe Ser Asn Gln Glu Val Tyr Ile Asp Arg Ile Glu Phe
580 585 590
Val Pro Ala Glu Val Thr Phe Glu Ala Glu Ser Asp Leu Glu Arg Ala
595 600 605
Gln Lys Ala Val Asn Glu Leu Phe Thr Ser Ser Asn Gln Ile Gly Leu
610 615 620
Lys Thr Asp Val Thr Asp Tyr His Ile Asp Gln Val Ser Asn Leu Val
625 630 635 640
Glu Cys Leu Ser Asp Glu Phe Cys Leu Asp Glu Lys Lys Glu Leu Ser
645 650 655
Glu Lys Val Lys His Ala Lys Arg Leu Ser Asp Glu Arg Asn Leu Leu
660 665 670
Gln Asp Pro Asn Phe Arg Gly Ile Asn Arg Gln Leu Asp Arg Gly Trp
675 680 685
Arg Gly Ser Thr Asp Ile Thr Ile Gln Gly Gly Asp Asp Val Phe Lys
690 695 700
Glu Asn Tyr Val Thr Leu Leu Gly Thr Phe Asp Glu Cys Tyr Pro Thr
705 710 715 720
Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Lys Leu Lys Ala Tyr Thr Arg
725 730 735
Tyr Gln Leu Arg Gly Tyr Ile Glu Asp Ser Gln Asp Leu Glu Ile Tyr
740 745 750
Leu Ile Arg Tyr Asn Ala Lys His Glu Thr Val Asn Val Pro Gly Thr
755 760 765
Gly Ser Leu Trp Pro Leu Ser Ala Pro Ser Pro Ile Gly Lys Cys Ala
770 775 780
His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys Thr Asp
785 790 795 800
Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys Thr Gln
805 810 815
Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu Lys Pro
820 825 830
Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys Lys Trp
835 840 845
Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val Tyr Lys
850 855 860
Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln Tyr Asp
865 870 875 880
Arg Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala Asp Lys
885 890 895
Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser Val Ile
900 905 910
Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg Ile Phe
915 920 925
Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn Gly Asp
930 935 940
Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val Asp Val
945 950 955 960
Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Val Pro Glu Trp Glu
965 970 975
Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly Tyr Ile
980 985 990
Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val Thr
995 1000 1005
Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser Asn
1010 1015 1020
Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn
1025 1030 1035
Asp Tyr Thr Ala Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser
1040 1045 1050
Arg Asn Arg Gly Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val
1055 1060 1065
Pro Ala Asp Tyr Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp
1070 1075 1080
Gly Arg Arg Asp Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp
1085 1090 1095
Tyr Thr Pro Leu Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr
1100 1105 1110
Phe Pro Glu Thr Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu
1115 1120 1125
Gly Thr Phe Ile Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1130 1135 1140
<210> 51
<211> 3513
<212> DNA
<213> Artificial
<220>
<223> Recombinant nucleotide sequence used for expression in a
bacterial cell encoding TIC836.
<400> 51
atggagaata atattcaaaa tcaatgcgta ccttacaatt gtttaaataa tcctgaagta 60
gaaatattaa atgaagaaag aagtactggc agattaccgt tagatatatc cttatcgctt 120
acacgtttcc ttttgagtga atttgttcca ggtgtgggag ttgcgtttgg attatttgat 180
ttaatatggg gttttataac tccttctgat tggagcttat ttcttttaca gattgaacaa 240
ttgattgagc aaagaataga aacattggaa aggaaccggg caattactac attacgaggg 300
ttagcagata gctatgaaat ttatattgaa gcactaagag agtgggaagc aaatcctaat 360
aatgcacaat taagggaaga tgtgcgtatt cgatttgcta atacagacga cgctttaata 420
acagcaataa ataattttac acttacaagt tttgaaatcc ctcttttatc ggtctatgtt 480
caagcggcga atttacattt atcactatta agagacgctg tatcgtttgg gcagggttgg 540
ggactggata tagctactgt taataatcat tataatagat taataaatct tattcataga 600
tatacgaaac attgtttgga cacatacaat caaggattag aaaacttaag aggtactaat 660
actcgacaat gggcaagatt caatcagttt aggagagatt taacacttac tgtattagat 720
atcgttgctc tttttccgaa ctacgatgtt agaacatatc caattcaaac gtcatcccaa 780
ttaacaaggg aaatttatac aagttcagta attgaggatt ctccagtttc tgctaatata 840
cctaatggtt ttaatagggc ggaatttgga gttagaccgc cccatcttat ggactttatg 900
aattctttgt ttgtaactgc agagactgtt agaagtcaaa ctgtgtgggg aggacactta 960
gttagttcac gaaatacggc tggtaaccgt ataaatttcc ctagttacgg ggtcttcaat 1020
cctggtggcg ccatttggat tgcagatgag gatccacgtc ctttttatcg gacattatca 1080
gatcctgttt ttgtccgagg aggatttggg aatcctcatt atgtactggg gcttagggga 1140
gtagcatttc aacaaactgg tacgaaccac acccgaacat ttagaaatag tgggaccata 1200
gattctctag atgaaatccc acctcaggat aatagtgggg caccttggaa tgattatagt 1260
catgtattaa atcatgttac atttgtacga tggccaggtg agatttcagg aagtgattca 1320
tggagagctc caatgttttc ttggacgcac cgtagtgcaa cccctacaaa tacaattgat 1380
ccggagagga ttacacaaat acctttaaca aaatctacta atcttggctc tggaacttct 1440
gtcgttaaag gaccaggatt tacaggagga gatattcttc gaagaacttc acctggccag 1500
atttcaacct taagagtaaa tattactgca ccattatcac aaagatatcg ggtaagaatt 1560
cgctacgctt ctaccacaaa tttacaattc catacatcaa ttgacggaag acctattaat 1620
caggggaatt tttcagcaac tatgagtagt gggagtaatt tacagtccgg aagctttagg 1680
actgtaggtt ttactactcc gtttaacttt tcaaatggat caagtgtatt tacgttaagt 1740
gctcatgtct tcaattcagg caatgaagtt tatatagatc gaattgaatt tgttccggca 1800
gaagtaacct ttgaggcaga atatgattta gaaagagcgc agaaggcggt gaatgcgctg 1860
tttacgtcta caaaccaact agggctaaaa acaaatgtaa cggattatca tattgatcaa 1920
gtgtccaatt tagttacgta tttatcggat gaattttgtc tggatgaaaa gcgagaattg 1980
tccgagaaag tcaaacatgc gaagcgactc agtgatgaac gcaatttact ccaagattca 2040
aatttcaaag acattaatag gcaaccagaa cgtgggtggg gcggaagtac agggattacc 2100
atccaaggag gggatgacgt atttaaagaa aattacgtca cactatcagg tacctttgat 2160
gagtgctatc caacatattt gtatcaaaaa atcgatgaat caaaattaaa agcctttacc 2220
cgttatcaat taagagggta tatcgaagat agtcaagact tagaaatcta tttaattcgc 2280
tacaatgcaa aacatgaaac agtaaatgtg ccaggtacgg gttccttatg gccgctttca 2340
gcccaaagtc caatcggaaa gtgtggagag ccgaatcgat gcgcgccaca ccttgaatgg 2400
aatcctgact tagattgttc gtgtagggat ggagaaaagt gtgcccatca ttcgcatcat 2460
ttctccttag acattgatgt aggatgtaca gacttaaatg aggacctagg tgtatgggtg 2520
atctttaaga ttaagacgca agatgggcac gcaagactag ggaatctaga gtttctcgaa 2580
gaaaaaccat tagtaggaga agcgctagct cgtgtgaaaa gagcggagaa aaaatggaga 2640
gacaaacgtg aaaaattgga atgggaaaca aatatcgttt ataaagaggc aaaagaatct 2700
gtagatgctt tatttgtaaa ctctcaatat gatcaattac aagcggatac gaatattgcc 2760
atgattcatg cggcagataa acgtgttcat agcattcgag aagcttatct gcctgagctg 2820
tctgtgattc cgggtgtcaa tgcggctatt tttgaagaat tagaagggcg tattttcact 2880
gcattctccc tatatgatgc gagaaatgtc attaaaaatg gtgattttaa taatggctta 2940
tcctgctgga acgtgaaagg gcatgtagat gtagaagaac aaaacaacca acgttcggtc 3000
cttgttgttc cggaatggga agcagaagtg tcacaagaag ttcgtgtctg tccgggtcgt 3060
ggctatatcc ttcgtgtcac agcgtacaag gagggatatg gagaaggttg cgtaaccatt 3120
catgagatcg agaacaatac agacgaactg aagtttagca actgcgtaga ggaggaaatc 3180
tatccaaata acacggtaac gtgtaatgat tatactgtaa atcaagaaga atacggaggt 3240
gcgtacactt ctcgtaatcg aggatataac gaagctcctt ccgtaccagc tgattatgcg 3300
tcagtctatg aagaaaaatc gtatacagat ggacgtagag agaatccttg tgaatttaac 3360
agagggtata gggattacac gccactacca gttggttatg tgacaaaaga attagaatac 3420
ttcccagaaa ccgataaggt atggattgag attggagaaa cggaaggaac atttatcgtg 3480
gacagcgtgg aattactcct tatggaggaa taa 3513
<210> 52
<211> 3513
<212> DNA
<213> Artificial
<220>
<223> Synthetic nucleotide sequence designed for expression in a plant
cell encoding TIC836.
<400> 52
atggagaaca acatccagaa ccagtgcgtg ccctacaact gcctgaacaa ccctgaggtt 60
gagatcctga acgaggagcg tagcaccggt aggctcccgc tagacatctc cctgagcctg 120
acccgcttcc tccttagtga gttcgtgccc ggcgtgggcg tggccttcgg cctcttcgac 180
ctcatctggg gcttcatcac tccttccgac tggtccctct tcctccttca gattgagcaa 240
ctgatcgagc agcgcatcga gacccttgag cgcaaccgcg ccatcaccac tctcagaggt 300
ctcgccgact cctacgaaat ctacatcgag gcactccgtg agtgggaggc caacccgaac 360
aatgcccagc tccgcgagga cgtgaggatc agattcgcca acaccgacga tgccctcatc 420
accgccatca acaatttcac cctcacctcc ttcgagatcc ctcttctgtc tgtgtacgtt 480
caagctgcta accttcacct ttccctcctg cgcgacgccg tgagcttcgg ccagggctgg 540
ggcctcgaca tcgccaccgt gaacaatcac tacaaccgcc tcatcaacct catccaccgc 600
tacaccaagc actgccttga cacctacaac cagggccttg agaacctccg tggcaccaac 660
acccgccagt gggcccgctt caaccagttc cgcagagacc tcaccctcac cgtgctcgac 720
atcgtggcac tcttcccaaa ctacgacgtg cgtacctacc ctatccagac ctccagccag 780
ctcaccaggg aaatctacac ctccagcgtg atcgaggact ctcctgtgtc cgccaacatc 840
cctaacggct tcaaccgcgc cgagttcggc gtgcgccctc ctcacctcat ggacttcatg 900
aactccctct tcgtcactgc cgagaccgtg cgctcccaga ccgtgtgggg cggtcacctc 960
gtgtccagcc gtaacaccgc tggcaacagg atcaacttcc cgtcctacgg cgtgttcaac 1020
ccaggcggtg ccatctggat cgccgatgaa gaccctcgtc ctttctaccg taccctgtcc 1080
gaccctgtgt tcgtgcgtgg cggtttcggc aaccctcact acgtgctggg cctgcgtggc 1140
gtggccttcc agcaaaccgg caccaaccac accaggacgt tccgtaactc cggcaccatc 1200
gacagtcttg acgagatccc tccgcaagac aactccggtg caccttggaa cgactactcc 1260
cacgtgctga accacgtgac cttcgtgagg tggcctggcg aaatctccgg ctccgactcc 1320
tggagggctc ctatgttcag ttggacccac aggagcgcta cgcctaccaa caccatcgac 1380
cctgagcgta tcactcagat ccctctgact aagagcacta acctgggcag cggcactagc 1440
gtggtcaagg gccctggctt cactggcggt gacatcctga ggcggactag ccctggccag 1500
atcagcactc tgagggtgaa catcactgct ccgctgagcc agcgttacag ggtcagaatc 1560
cgttacgctt ctactactaa ccttcagttc cacactagca tcgacggccg tccgatcaac 1620
cagggcaact tctctgctac tatgagttct ggcagtaacc tccagtctgg tagtttccgg 1680
actgtcggtt tcactacgcc gttcaacttc tccaacggta gttctgtctt cactctgtct 1740
gctcacgtgt tcaactctgg caacgaggtg tacatcgacc ggatcgagtt cgtccctgct 1800
gaggtgacgt tcgaggccga gtacgacctg gagcgggctc agaaggctgt caacgctctg 1860
ttcacttcta ctaaccagct tggtttgaag actaacgtga ccgactacca cattgatcaa 1920
gtcagtaacc tggtcacgta cctgtctgac gagttctgtc ttgacgagaa gcgggagctg 1980
tctgagaagg tcaagcacgc taagcggctg tctgacgagc ggaacctgct tcaagacagt 2040
aacttcaagg acattaaccg ccagcctgag cgtggttggg gagggtccac gggtattacg 2100
attcaaggag gtgacgatgt ctttaaggag aactatgtga cgctttcggg tacgtttgat 2160
gagtgctatc caacgtacct ttaccagaag attgacgagt cgaagctgaa ggctttcact 2220
cgttaccagc ttcgtggtta cattgaggac tcgcaagacc tcgaaatcta cctcattcgt 2280
tacaacgcta agcacgagac tgtcaacgtc cctggtacgg gtagtctttg gccgctttct 2340
gctcagtcgc cgattggcaa gtgtggcgag ccgaaccgtt gcgctcctca cttggagtgg 2400
aacccggatc tcgattgctc gtgccgtgac ggtgagaagt gcgcgcacca tagtcatcac 2460
tttagccttg acattgatgt cggttgcacg gatcttaacg aggatcttgg agtctgggtg 2520
attttcaaga tcaaaactca ggatgggcac gcgcgtcttg ggaatcttga gttcctggag 2580
gagaagccac ttgtcggtga ggcgcttgcg cgtgtcaagc gtgcggagaa gaaatggcgt 2640
gataagcgtg agaagttgga gtgggagacg aacatcgtgt acaaggaggc gaaggagtcg 2700
gtcgatgcgt tgtttgtcaa tagtcaatac gatcaattgc aagcggatac gaacatcgca 2760
atgattcatg cggcagataa gcgtgtccat tcgattcgtg aggcgtactt gccagagttg 2820
tcggtcatcc caggagttaa tgcggcaatc tttgaggaat tggagggcag aatcttcacg 2880
gcgttctcgt tgtacgatgc aagaaatgtt attaagaatg gagatttcaa caatgggttg 2940
tcatgctgga atgttaaggg tcacgttgat gttgaagaac agaacaacca gagatcagtg 3000
ttggttgtac cagagtggga ggcagaggtt tcacaagagg tgagagtttg cccaggcaga 3060
ggctacatct tgagagttac agcatacaaa gagggatacg gcgagggatg tgttacaatc 3120
cacgaaatcg agaacaatac cgatgagcta aagttctcaa attgtgttga ggaggagatc 3180
tacccgaaca acacggttac ttgtaatgat tacacagtga accaggagga gtatggtggt 3240
gcatacacat caagaaatag aggctacaat gaagcaccat cagttccagc agattatgcc 3300
tcagtttatg aggagaagtc atacacagat ggacgacgtg agaatccatg tgagttcaat 3360
cgaggatacc gagattacac accactacca gttggatacg ttacaaagga actagaatac 3420
ttcccagaaa cagataaagt atggatagag atcggagaaa cagaaggaac attcatcgtt 3480
gattcagtag aactactact tatggaagaa tga 3513
<210> 53
<211> 1170
<212> PRT
<213> Artificial
<220>
<223> Amino acid sequence of the chimeric protein TIC836.
<400> 53
Met Glu Asn Asn Ile Gln Asn Gln Cys Val Pro Tyr Asn Cys Leu Asn
1 5 10 15
Asn Pro Glu Val Glu Ile Leu Asn Glu Glu Arg Ser Thr Gly Arg Leu
20 25 30
Pro Leu Asp Ile Ser Leu Ser Leu Thr Arg Phe Leu Leu Ser Glu Phe
35 40 45
Val Pro Gly Val Gly Val Ala Phe Gly Leu Phe Asp Leu Ile Trp Gly
50 55 60
Phe Ile Thr Pro Ser Asp Trp Ser Leu Phe Leu Leu Gln Ile Glu Gln
65 70 75 80
Leu Ile Glu Gln Arg Ile Glu Thr Leu Glu Arg Asn Arg Ala Ile Thr
85 90 95
Thr Leu Arg Gly Leu Ala Asp Ser Tyr Glu Ile Tyr Ile Glu Ala Leu
100 105 110
Arg Glu Trp Glu Ala Asn Pro Asn Asn Ala Gln Leu Arg Glu Asp Val
115 120 125
Arg Ile Arg Phe Ala Asn Thr Asp Asp Ala Leu Ile Thr Ala Ile Asn
130 135 140
Asn Phe Thr Leu Thr Ser Phe Glu Ile Pro Leu Leu Ser Val Tyr Val
145 150 155 160
Gln Ala Ala Asn Leu His Leu Ser Leu Leu Arg Asp Ala Val Ser Phe
165 170 175
Gly Gln Gly Trp Gly Leu Asp Ile Ala Thr Val Asn Asn His Tyr Asn
180 185 190
Arg Leu Ile Asn Leu Ile His Arg Tyr Thr Lys His Cys Leu Asp Thr
195 200 205
Tyr Asn Gln Gly Leu Glu Asn Leu Arg Gly Thr Asn Thr Arg Gln Trp
210 215 220
Ala Arg Phe Asn Gln Phe Arg Arg Asp Leu Thr Leu Thr Val Leu Asp
225 230 235 240
Ile Val Ala Leu Phe Pro Asn Tyr Asp Val Arg Thr Tyr Pro Ile Gln
245 250 255
Thr Ser Ser Gln Leu Thr Arg Glu Ile Tyr Thr Ser Ser Val Ile Glu
260 265 270
Asp Ser Pro Val Ser Ala Asn Ile Pro Asn Gly Phe Asn Arg Ala Glu
275 280 285
Phe Gly Val Arg Pro Pro His Leu Met Asp Phe Met Asn Ser Leu Phe
290 295 300
Val Thr Ala Glu Thr Val Arg Ser Gln Thr Val Trp Gly Gly His Leu
305 310 315 320
Val Ser Ser Arg Asn Thr Ala Gly Asn Arg Ile Asn Phe Pro Ser Tyr
325 330 335
Gly Val Phe Asn Pro Gly Gly Ala Ile Trp Ile Ala Asp Glu Asp Pro
340 345 350
Arg Pro Phe Tyr Arg Thr Leu Ser Asp Pro Val Phe Val Arg Gly Gly
355 360 365
Phe Gly Asn Pro His Tyr Val Leu Gly Leu Arg Gly Val Ala Phe Gln
370 375 380
Gln Thr Gly Thr Asn His Thr Arg Thr Phe Arg Asn Ser Gly Thr Ile
385 390 395 400
Asp Ser Leu Asp Glu Ile Pro Pro Gln Asp Asn Ser Gly Ala Pro Trp
405 410 415
Asn Asp Tyr Ser His Val Leu Asn His Val Thr Phe Val Arg Trp Pro
420 425 430
Gly Glu Ile Ser Gly Ser Asp Ser Trp Arg Ala Pro Met Phe Ser Trp
435 440 445
Thr His Arg Ser Ala Thr Pro Thr Asn Thr Ile Asp Pro Glu Arg Ile
450 455 460
Thr Gln Ile Pro Leu Thr Lys Ser Thr Asn Leu Gly Ser Gly Thr Ser
465 470 475 480
Val Val Lys Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr
485 490 495
Ser Pro Gly Gln Ile Ser Thr Leu Arg Val Asn Ile Thr Ala Pro Leu
500 505 510
Ser Gln Arg Tyr Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu
515 520 525
Gln Phe His Thr Ser Ile Asp Gly Arg Pro Ile Asn Gln Gly Asn Phe
530 535 540
Ser Ala Thr Met Ser Ser Gly Ser Asn Leu Gln Ser Gly Ser Phe Arg
545 550 555 560
Thr Val Gly Phe Thr Thr Pro Phe Asn Phe Ser Asn Gly Ser Ser Val
565 570 575
Phe Thr Leu Ser Ala His Val Phe Asn Ser Gly Asn Glu Val Tyr Ile
580 585 590
Asp Arg Ile Glu Phe Val Pro Ala Glu Val Thr Phe Glu Ala Glu Tyr
595 600 605
Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe Thr Ser Thr
610 615 620
Asn Gln Leu Gly Leu Lys Thr Asn Val Thr Asp Tyr His Ile Asp Gln
625 630 635 640
Val Ser Asn Leu Val Thr Tyr Leu Ser Asp Glu Phe Cys Leu Asp Glu
645 650 655
Lys Arg Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg Leu Ser Asp
660 665 670
Glu Arg Asn Leu Leu Gln Asp Ser Asn Phe Lys Asp Ile Asn Arg Gln
675 680 685
Pro Glu Arg Gly Trp Gly Gly Ser Thr Gly Ile Thr Ile Gln Gly Gly
690 695 700
Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Ser Gly Thr Phe Asp
705 710 715 720
Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu Ser Lys Leu
725 730 735
Lys Ala Phe Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu Asp Ser Gln
740 745 750
Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His Glu Thr Val
755 760 765
Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala Gln Ser Pro
770 775 780
Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His Leu Glu Trp
785 790 795 800
Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys Cys Ala His
805 810 815
His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys Thr Asp Leu
820 825 830
Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys Thr Gln Asp
835 840 845
Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu Lys Pro Leu
850 855 860
Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys Lys Trp Arg
865 870 875 880
Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val Tyr Lys Glu
885 890 895
Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln Tyr Asp Gln
900 905 910
Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala Asp Lys Arg
915 920 925
Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser Val Ile Pro
930 935 940
Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg Ile Phe Thr
945 950 955 960
Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn Gly Asp Phe
965 970 975
Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val Asp Val Glu
980 985 990
Glu Gln Asn Asn Gln Arg Ser Val Leu Val Val Pro Glu Trp Glu Ala
995 1000 1005
Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly Tyr Ile
1010 1015 1020
Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys Val
1025 1030 1035
Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser
1040 1045 1050
Asn Cys Val Glu Glu Glu Ile Tyr Pro Asn Asn Thr Val Thr Cys
1055 1060 1065
Asn Asp Tyr Thr Val Asn Gln Glu Glu Tyr Gly Gly Ala Tyr Thr
1070 1075 1080
Ser Arg Asn Arg Gly Tyr Asn Glu Ala Pro Ser Val Pro Ala Asp
1085 1090 1095
Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg
1100 1105 1110
Glu Asn Pro Cys Glu Phe Asn Arg Gly Tyr Arg Asp Tyr Thr Pro
1115 1120 1125
Leu Pro Val Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu
1130 1135 1140
Thr Asp Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe
1145 1150 1155
Ile Val Asp Ser Val Glu Leu Leu Leu Met Glu Glu
1160 1165 1170
Claims (22)
- 서열번호 28을 포함하는 키메릭 살곤충 단백질(chimeric insecticidal protein).
- 제1항에 있어서, 상기 키메릭 살곤충 단백질이 인시목 곤충(order Lepidoptera) 종에 대해서 저해 활성을 나타내는, 키메릭 살곤충 단백질.
- 제2항에 있어서, 상기 곤충 종이 안티카르시아 겜마탈리스(Anticarsia gemmatalis), 디아트라에아 사카랄리스(Diatraea saccharalis), 엘라스모팔푸스 리그노셀루스(Elasmopalpus lignosellus), 헬리코베르파 제아(Helicoverpa zea), 헬리오티스 비레센스(Heliothis virescens), 크리소데익시스 인클루덴스(Chrysodeixis includens), 스포돕테라 코스미오이데스(Spodoptera cosmioides), 스포돕테라 에리다니아(Spodoptera eridania), 스포돕테라 프루기페르다(Spodoptera frugiperda), 스포돕테라 엑시구아(Spodoptera exigua), 헬리코베르파 아르미게라(Helicoverpa armigera), 스포돕테라 리투라(Spodoptera litura), 펙티노포라 고시피엘라(Pectinophora gossypiella), 디아트라에아 그란디오셀라(Diatraea grandiosella), 에아리아스 비텔라(Earias vitella), 헬리코베르파 겔로토페온(Helicoverpa gelotopeon), 및 라치플루시아 누(Rachiplusia nu)로 이루어진 군으로부터 선택되는, 키메릭 살곤충 단백질.
- 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드로서, 상기 폴리뉴클레오타이드가 이종 프로모터(heterologous promoter)에 작동 가능하게 연결되어 있고, 상기 키메릭 살곤충 단백질이 서열번호 28을 포함하는, 폴리뉴클레오타이드.
- 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드로서, 상기 폴리뉴클레오타이드는,
a. 서열번호 27을 포함하거나; 또는
b. 서열번호 28을 암호화하는 뉴클레오타이드 서열을 포함하는, 폴리뉴클레오타이드. - 숙주 세포로서, 서열번호 27을 포함하되, 상기 숙주 세포가 박테리아 숙주 세포 또는 식물 숙주 세포로 이루어진 군으로부터 선택되는, 숙주 세포.
- 제6항에 있어서, 박테리아 숙주 세포가 아그로박테리움(Agrobacterium), 리조븀(Rhizobium), 바실러스(Bacillus), 브레비바실러스(Brevibacillus), 에쉐리키아(Escherichia), 슈도모나스(Pseudomonas), 클렙시엘라(Klebsiella), 및 에르위니아(Erwinia)로 이루어진 군으로부터 선택되는, 숙주 세포.
- 제6항에 있어서, 식물 숙주 세포가 외떡잎 식물 및 쌍떡잎 식물로 이루어진 식물 군으로부터 선택되는, 숙주 세포.
- 서열번호 28을 포함하는 키메릭 살곤충 단백질을 포함하는, 곤충 저해 조성물.
- 제9항에 있어서, 키메릭 살곤충 단백질과 상이한 적어도 1종의 곤충 저해제를 추가로 포함하는, 곤충 저해 조성물.
- 제10항에 있어서, 적어도 1종의 곤충 저해제가 곤충 저해 단백질, 곤충 저해 dsRNA 분자, 및 곤충 저해 화학물질로 이루어진 군으로부터 선택되는, 곤충 저해 조성물.
- 제10항에 있어서, 적어도 1종의 다른 살충제가 인시목, 딱정벌레목, 노린재목, 동시아목, 또는 총채벌레목 중 1종 이상의 해충 종에 대해서 활성을 나타내는, 곤충 저해 조성물.
- 곤충 저해 유효량의
a. 서열번호 28인 아미노산 서열을 포함하는 키메릭 살곤충 단백질; 또는
b. 서열번호 27을 포함하는 폴리뉴클레오타이드를 포함하는 종자. - 인시류 해충을 억제량의 제1항의 키메릭 살곤충 단백질과 접촉시키는 것을 포함하는, 인시류 해충의 방제 방법.
- 서열번호 28을 포함하는, 트랜스제닉 식물 또는 식물 부분.
- 제15항의 트랜스제닉 식물 또는 식물 부분이 인시류 억제량의 키메릭 살곤충 단백질을 발현하는, 해충을 상기 식물 또는 식물 부분에 노출시키는 것을 포함하는, 인시류 해충의 방제 방법.
- 검출가능한 양의 키메릭 살곤충 단백질을 포함하는, 제15항의 식물 또는 식물 부분으로부터 유래된 상품(commodity product).
- 제17항에 있어서, 식물 바이오매스(biomass), 오일, 곡물(meal), 동물 사료, 곡물 가루, 플레이크, 겨(bran), 린트, 외피, 및 가공된 종자로 이루어진 군으로부터 선택된, 상품.
- 제1항의 키메릭 살곤충 단백질을 포함하는 종자의 생산 방법으로서,
a. 제1항의 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드를 포함하는 적어도 하나의 종자를 심는 단계;
b. 종자로부터 식물을 성장시키는 단계; 및
c. 식물로부터 종자를 수확하는 단계를 포함하고, 수확된 종자는 제1항의 키메릭 살곤충 단백질을 암호화하는 상기 폴리뉴클레오타이드를 포함하는, 종자의 생산 방법. - 키메릭 살곤충 단백질을 암호화하는 폴리뉴클레오타이드 분절에 작동 가능하게 연결된 이종 프로모터를 포함하는 재조합 핵산 분자로서,
a. 상기 키메릭 살곤충 단백질이 서열번호 28을 포함하거나; 또는
b. 상기 폴리뉴클레오타이드 분절이 서열번호 27을 포함하는, 재조합 핵산 분자. - 서열번호 28을 포함하는 트렌스제닉 식물 세포.
- 제21항의 트렌스제닉 식물 세포에 인시류 해충을 노출시키는 것을 포함하는 인시류 해충을 방제하는 방법으로서, 상기 식물 세포는 키메릭 살곤충 단백질의 인시류 억제량을 발현하는, 방법.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462064989P | 2014-10-16 | 2014-10-16 | |
| US62/064,989 | 2014-10-16 | ||
| PCT/US2015/055800 WO2016061391A2 (en) | 2014-10-16 | 2015-10-15 | Novel chimeric insecticidal proteins toxic or inhibitory to lepidopteran pests |
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| Application Number | Title | Priority Date | Filing Date |
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| KR1020177013031A Division KR102127553B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
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| Publication Number | Publication Date |
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| KR20190142456A true KR20190142456A (ko) | 2019-12-26 |
| KR102208985B1 KR102208985B1 (ko) | 2021-01-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| KR1020197037524A KR102208984B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
| KR1020197037522A KR102208978B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
| KR1020197037525A KR102208985B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
| KR1020177013031A KR102127553B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
| KR1020197037523A KR102208980B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
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| KR1020197037522A KR102208978B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
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| KR1020197037523A KR102208980B1 (ko) | 2014-10-16 | 2015-10-15 | 인시류 해충에 대해서 유독성이거나 저해성인 신규한 키메릭 살곤충 단백질 |
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| US (7) | US10233217B2 (ko) |
| EP (5) | EP3715362A1 (ko) |
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| KR (5) | KR102208984B1 (ko) |
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| UA120843C2 (uk) * | 2013-12-09 | 2020-02-25 | Атенікс Корп. | Конструкція, яка містить гетерологічиий промотор, функціонально зв'язаний із нуклеотидною послідовністю, яка кодує амінокислотну послідовність, що має пестицидну активність до лускокрилих |
| NZ768153A (en) | 2014-10-16 | 2023-12-22 | Monsanto Technology Llc | Novel chimeric insecticidal proteins toxic or inhibitory to lepidopteran pests |
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| WO2002014517A1 (en) * | 2000-08-11 | 2002-02-21 | Monsanto Technology Llc | BROAD-SPECTRUM δ-ENDOTOXINS |
| KR20120096571A (ko) * | 2009-12-16 | 2012-08-30 | 다우 아그로사이언시즈 엘엘씨 | 곤충 내성 관리를 위한 ＣＲＹ１Ｃａ 및 ＣＲＹ１Ｆａ 단백질의 조합 용도 |
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