[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

CN114058689B - 一种基因突变检测试剂盒及其应用 - Google Patents

一种基因突变检测试剂盒及其应用 Download PDF

Info

Publication number
CN114058689B
CN114058689B CN202010754298.0A CN202010754298A CN114058689B CN 114058689 B CN114058689 B CN 114058689B CN 202010754298 A CN202010754298 A CN 202010754298A CN 114058689 B CN114058689 B CN 114058689B
Authority
CN
China
Prior art keywords
artificial sequence
group
mut
base extension
dna
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010754298.0A
Other languages
English (en)
Other versions
CN114058689A (zh
Inventor
许争峰
胡平
谭建新
姜正文
余锋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
GENESKY DIAGNOSTICS (SUZHOU) Inc
Nanjing Maternity and Child Healthcare Hospital
Original Assignee
GENESKY DIAGNOSTICS (SUZHOU) Inc
Nanjing Maternity and Child Healthcare Hospital
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by GENESKY DIAGNOSTICS (SUZHOU) Inc, Nanjing Maternity and Child Healthcare Hospital filed Critical GENESKY DIAGNOSTICS (SUZHOU) Inc
Priority to CN202010754298.0A priority Critical patent/CN114058689B/zh
Publication of CN114058689A publication Critical patent/CN114058689A/zh
Application granted granted Critical
Publication of CN114058689B publication Critical patent/CN114058689B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/16—Primer sets for multiplex assays
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

本发明提供了一种ATP7B、PAH、PTS、MUT、MMACHC基因突变检测试剂盒及其应用。具体地，本发明利用SNaPshot技术对ATP7B、PAH、PTS、MUT、MMACHC基因的224个热点突变进行检测。设计了51对PCR引物用于扩增了包含224个位点的51个100‑500bp片段，也设计了224个紧邻目标位点的延伸引物用于单碱基延伸。本发明的试剂盒具有非常的高的准确性，能够快速进行相关疾病的基因诊断，为临床医生提供有效参考。

Description

一种基因突变检测试剂盒及其应用
技术领域
本发明涉及遗传学领域，更具体地涉及一种基因突变检测试剂盒及其应用。
背景技术
遗传代谢疾病是基因突变导致维持机体正常代谢所必需的某些酶、受体或载体缺乏或蛋白质异常发生的疾病，导致出现相应的病理改变和临床表现。尽管单一遗传代谢病的发病率低，但总体发病却能达到活产婴儿的1/500。目前，遗传代谢病的基因诊断主要基于二代测序技术。然而，二代测序样品制备繁琐，产生的数据量较大，对后期的数据的拼接和生物信息学分析要求较高，不适合大规模推广。此外，尽管随着二代测序技术取得较大的发展，但其的价格仍然相对较高。因此，临床上亟需一种操作简便、易于解读、价格较低的遗传代谢疾病快速基因检测方法。
发明内容
本发明的目的在于提供一种基因突变检测试剂盒及其应用。
本发明的的目的在于在建立一种操作简便、易于解读、价格较低的用于包含肝豆状核变性致病基因ATP7B、苯丙酮尿症致病基因PAH和PTS，以及甲基丙二酸血症的致病基因MUT和MMACHC的224个突变的快速检测方法。
在本发明的第一方面，提供了一种基因突变检测试剂盒，所述试剂盒包含特异性检测ATP7B基因的突变位点的核酸检测试剂；并且，所述ATP7B基因的突变位点选自下组A：
组A
在另一优选例中，所述ATP7B基因的突变位点选自下组B：
组B
在另一优选例中，所述试剂盒还包含特异性检测MMACHC基因的突变位点的核酸检测试剂，并且，所述MMACHC基因的突变位点选自下组C：
组C
在另一优选例中，所述试剂盒还包含特异性检测MUT基因的突变位点的核酸检测试剂，并且，所述MUT基因的突变位点选自下组D：
组D
在另一优选例中，所述试剂盒还包含特异性检测PAH基因和PTS基因的突变位点的核酸检测试剂，并且，所述PAH基因和PTS基因的突变位点选自下组E：
组E
在另一优选例中，所述PAH基因和PTS基因的突变位点选自下组F：
组F
在另一优选例中，所述PAH基因和PTS基因的突变位点选自下组G：
组G
在另一优选例中，所述试剂盒中包含的特异性检测组A、组B、组C、组D、组E、组F、组G的突变位点的核酸检测试剂位于不同的容器中。
在另一优选例中，所述试剂盒包含针对所述突变位点的特异性多重PCR引物。
在另一优选例中，所述多重PCR引物对通过扩增能够同时得到包含不同突变位点的扩增子，每段扩增子至少含有1-5个突变位点。
在另一优选例中，所述多重PCR引物的序列如SEQ ID NO:1-103所示。
在另一优选例中，所述试剂盒包含针对ATP7B基因(组A和组B的突变位点)的多重PCR引物，并且，所述多重PCR引物的序列如SEQ ID NO:1-32所示。
在另一优选例中，所述试剂盒包含针对MMACHC基因(组C的突变位点)的多重PCR引物，并且，所述多重PCR引物的序列如SEQ ID NO:33-40所示。
在另一优选例中，所述试剂盒包含针对MUT基因(组D的突变位点)的多重PCR引物，并且，所述多重PCR引物的序列如SEQ ID NO:41-62所示。
在另一优选例中，所述试剂盒包含针对PAH基因和PTS基因(组E、组F和组G的突变位点)的多重PCR引物，并且，所述多重PCR引物的序列如SEQ ID NO:63-103所示。
在另一优选例中，所述试剂盒还包含针对所述突变位点的单碱基延伸引物。
在另一优选例中，所述单碱基延伸引物为SNaPshot单碱基延伸引物。
在另一优选例中，所述试剂盒包含针对组A的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:104-136所示。
在另一优选例中，所述试剂盒包含针对组B的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:137-171所示。
在另一优选例中，所述试剂盒包含针对组C的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:172-190所示。
在另一优选例中，所述试剂盒包含针对组D的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:191-215所示。
在另一优选例中，所述试剂盒包含针对组E的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:216-251所示。
在另一优选例中，所述试剂盒包含针对组F的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:252-288所示。
在另一优选例中，所述试剂盒包含针对组G的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ ID NO:289-321所示。
在另一优选例中，所述试剂盒还包含所述试剂盒中还包括标准品DNA。
在另一优选例中，所述试剂盒还包含测序酶，较佳地，所述测序酶包括snapshot测序酶。
在另一优选例中，所述试剂盒还包含荧光标记的ddNTP。
在本发明的第二方面，提供了一种检测来自ATP7B、PAH、PTS、MUT和MMACHC基因的突变位点的方法，包括步骤：
(i)提供一待测样品；
(ii)提供如本发明第一方面所述的试剂盒，对待测样品进行多重PCR反应以及单碱基延伸反应；
(iii)对步骤(ii)所获得的样品进行检测和分型。
在另一优选例中，所述单碱基延伸反应为SNaPshot单碱基延伸反应。
在另一优选例中，所述方法还包括对对照样品进行平行检测。
在另一优选例中，所述对照样品包括阳性对照样品和阴性对照样品。
在另一优选例中，所述阴性对照样品为超纯水。
在另一优选例中，在所述步骤(ii)中，待测样品进行复合扩增反应之后，在进行单碱基延伸反应之前，用虾碱性磷酸酶(SAP)进行纯化处理。
在另一优选例中，在所述步骤(iii)之前，对步骤(ii)所获得的样品进行虾碱性磷酸酶(SAP)纯化处理。
在另一优选例中，在所述步骤(iii)中，通过毛细管电泳进行检测。
在另一优选例中，在所述步骤(iii)中，通过质谱法进行检测。
在另一优选例中，所述的方法是非诊断和非治疗性的。
在另一优选例中，所述的样本是含人体DNA的样本。
在本发明的第三方面，提供了一种如本发明第一方面所述的试剂盒的用途，用于检测来自ATP7B、PAH、PTS、MUT和MMACHC基因的突变位点。
在另一优选例中，所述试剂盒用于选自下组的疾病的基因诊断：
肝豆状核变性、苯丙酮尿症、四氢生物蝶呤缺乏症、甲基丙二酸血症、或其组合。
在本发明的第四方面，提供了一种核酸检测试剂的用途，用于制备基因突变检测试剂盒，其中，所述的核酸检测试剂包括特异性检测ATP7B基因的突变位点核酸检测试剂；并且，所述ATP7B基因的突变位点选自组A。
在另一优选例中，所述ATP7B基因的突变位点选自组B。
在另一优选例中，所述的核酸检测试剂包括特异性检测MMACHC基因的突变位点的核酸检测试剂，并且，所述MMACHC基因的突变位点选自组C。
在另一优选例中，所述的核酸检测试剂包括特异性检测MUT基因的突变位点的核酸检测试剂，并且，所述MUT基因的突变位点选自组D。
在另一优选例中，所述的核酸检测试剂包括特异性检测PAH基因和PTS基因的突变位点的核酸检测试剂，并且，所述PAH基因和PTS基因的突变位点选自组E。
在另一优选例中，所述PAH基因和PTS基因的突变位点选自下组F。
在另一优选例中，所述PAH基因和PTS基因的突变位点选自下组G。
在另一优选例中，所述基因突变检测试剂盒用于检测ATP7B、PAH、PTS、MUT和/或MMACHC基因的突变。
在另一优选例中，所述基因突变检测试剂盒用于选自下组的疾病的基因诊断：
肝豆状核变性、苯丙酮尿症、四氢生物蝶呤缺乏症、甲基丙二酸血症、或其组合。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1显示了本发明检测方法的工作原理。
图2显示了一例阴性样本ATP7B-Panel1的毛细管电泳峰形图，包含ATP7B基因33个热点突变。
图3显示了一例阴性样本ATP7B-Panel2的毛细管电泳峰形图，包含ATP7B基因37个热点突变。
图4显示了一例阴性样本PKU-Panel1的毛细管电泳峰形图，包含PAH基因34个热点突变，PTS基因4个热点突变。
图5显示了一例阴性样本PKU-Panel2的毛细管电泳峰形图，包含PAH基因32个热点突变，PTS基因6个热点突变。
图6显示了一例阴性样本PKU-Panel3的毛细管电泳峰形图，包含PAH基因31个热点突变，PTS基因3个热点突变。
图7显示了一例阴性样本MUT的毛细管电泳峰形图，包含MUT基因的25个热点突变。
图8显示了一例阴性样本MMACHC的毛细管电泳峰形图，包含MMACHC基因的19个热点突变。
图9显示了ATP7B:c.2621C>T杂合突变的毛细管电泳峰形图(上图为杂合突变样本，下图为阴性样本)。
图10显示了ATP7B:c.2621C>T杂合突变的一代测序验证图。
图11显示了MMACHC:c.217C>T杂合突变的毛细管电泳峰形图(上图为杂合突变样本，下图为阴性样本)。
图12显示了MMACHC:c.217C>T杂合突变的一代测序验证图。
图13显示了MUT:c.729_730insTT杂合突变的毛细管电泳峰形图(上图为杂合突变样本，下图为阴性样本)。
图14显示MUT:c.729_730insTT杂合突变的一代测序验证图。
图15显示了PAH:c.1033G>A杂合突变的毛细管电泳峰形图(上图为杂合突变样本，下图为阴性样本)。
图16显示PAH:c.1033G>A杂合突变的一代测序验证图。
具体实施方式
本发明人经过广泛而深入地研究，首次意外地发现一种ATP7B、PAH、PTS、MUT、MMACHC基因突变检测试剂盒及其应用。具体地，本发明利用SNaPshot技术对ATP7B、PAH、PTS、MUT、MMACHC基因的224个热点突变进行检测。设计了51对PCR引物用于扩增了包含224个位点的51个100-500bp片段，也设计了224个紧邻目标位点的延伸引物用于单碱基延伸。PCR产物用Qiagen公司的HotStarTaq进行多重PCR获得，PCR产物经虾碱酶(SAP)(fromPromega)和外切酶I(EXO I)(from Epicentre)纯化后用ABI公司的SNaPshot Multiplexkit进行延伸反应。延伸产物用虾碱酶(SAP)(from Promega)纯化后在ABI3730xl上样。SNP分型用GeneMapper4.0(Appliedbiosystems)来分析。
本发明的主要的工作原理如图1所示，在一个含有测序酶、四种荧光标记的ddNTP、紧邻多态位点5’端的不同长度的延伸引物和PCR模板的反应体系中，引物延伸一个碱基即终止，经ABI测序仪毛细管电泳后，根据峰的颜色可知掺入的碱基种类，从而确定该样本的基因型，根据峰图上峰移动的位置确定该延伸产物对应的突变位点。
本发明的主要优点包括：
(a)本试剂盒在筛查的遗传性疾病涵盖上有所突破。
(b)本试剂盒检测了肝豆状核变性、苯丙酮尿症、四氢生物蝶呤缺乏症、甲基丙二酸血症相关基因ATP7B、PAH、PTS、MUT、MMACHC的224个热点突变，能快速进行相关疾病的基因诊断，为临床医生提供有效参考。
(c)本技术采用具有高准确性的SNaPshot技术，具有非常的高的准确性。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，例如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring HarborLaboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。
通用材料
实施例中的DNA样本获自南京市妇幼保健院，分离自人外周血。
实施例中涉及的突变位点信息以及对应的引物和分组如表1-4所示。
表1突变位点信息
表2：PCR引物序列表
表3 PCR引物和各个突变位点的对应关系
表4延伸引物信息
实施例1突变位点的检测
采用SNaPshot技术，对肝豆状核变性、苯丙酮尿症、四氢生物蝶呤缺乏症、甲基丙二酸血症相关基因ATP7B、PAH、PTS、MUT、MMACHC的224个热点突变进行检测，具体实验操作步骤如下：
1)DNA样本取1μl 1％琼脂糖凝胶电泳对其样本进行质量检查以及浓度估计，根据估计的浓度将样本稀释到工作浓度5-10ng/μl.
2)PCR反应
反应体系(10μl)：
PCR循环程序：
95℃ 5min；11×(94℃ 20s,65℃-0.5℃/循环40s,72℃ 1.5min)；24×(94℃20s,59℃ 30s,72℃ 1.5min)；72℃ 2min；4℃保温.
3)PCR产物纯化
在10μl PCR产物中加入1U SAP酶和1U Exonuclease I酶，37℃温育1小时,然后75℃灭活15分钟。
4)SNaPshot多重单碱基延伸反应
延伸反应体系(10μl)：
PCR循环程序：
96℃ 1min；28×(96℃ 10s,50℃ 5s,60℃ 30s)；4℃保温
5)延伸产物纯化
在10μl延伸产物中加入1U SAP酶，37℃温育1小时,然后75℃灭活15分钟.
6)延伸产物上ABI3730XL测序仪
取0.5μl纯化后的延伸产物，与0.5μl Liz120 SIZE STANDARD，9μl Hi-Di混匀，95℃变性5分钟后上ABI3730XL测序仪
7)ABI3730XL测序仪上收集的原始数据用GeneMapper 4.0(AppliedBiosystemsCo.,Ltd.,USA)进行分析。
本实施例共检测了5375例样本，其中ATP7B基因阳性有119例、MMACHC基因阳性有61例、MUT基因阳性有14例、PAH基因阳性有97例、PTS基因阳性有27例。随后，通过SANGER测序对检测为阳性的样本进行结果验证，其正确率达到了100％。
部分阳性突变样本的检测结果及其阴性对照和验证结果如图9-16所示。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
序列表
<110> 南京市妇幼保健院
天昊生物医药科技（苏州）有限公司
<120> 一种基因突变检测试剂盒及其应用
<130> P2020-1404
<160> 321
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
gccaccagca cagtcaggat ct 22
<210> 2
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
ccaggcttaa gggagccact tt 22
<210> 3
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
gacccttctt gtaccagccc agt 23
<210> 4
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
caagctcagg acatgcctca aa 22
<210> 5
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
agtgctggga attccatggt g 21
<210> 6
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
cgaggtctat acgcagcatt cc 22
<210> 7
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
ggctttcaca ggctttcctt gat 23
<210> 8
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
ccatgggaaa agttgaagaa ttttg 25
<210> 9
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
gtcctggacc acaacatcat tcc 23
<210> 10
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
aatatctgag ggccacacac agc 23
<210> 11
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
tacttgctgg cagccttcac tg 22
<210> 12
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ggagcagctc ttttctgaac ctg 23
<210> 13
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
tcatgtctct ccaagccaca ga 22
<210> 14
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
gtctctgccc acactcacaa gg 22
<210> 15
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
gtgataagtg gcgtttgttg cag 23
<210> 16
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ttccaccagt ttcacaatct gagc 24
<210> 17
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
cccaatcttt atccatgctt gtgg 24
<210> 18
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
cacaaccacc atatagccca agg 23
<210> 19
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
tttcagaacc ccaacaagca cat 23
<210> 20
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
tggctctcag gcttttctct caa 23
<210> 21
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
ggaagcccct ccatctgtat tg 22
<210> 22
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
ggagagaagg acatggtgag gaa 23
<210> 23
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
cacttcaccc ctcttggctt aca 23
<210> 24
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
caataccttt ttctgcggga agg 23
<210> 25
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
ggatgctgtc acaagaggtg ctt 23
<210> 26
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
tttctgagag agcggaagga agg 23
<210> 27
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
ccccaacttg tgtagctgct gat 23
<210> 28
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
ctgggccaac tggtgcttac ttt 23
<210> 29
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
ccttttgcca acactaggca ttg 23
<210> 30
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
cagctggagc acagtgggta aga 23
<210> 31
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
cctcctctgt gtctgtggtg ct 22
<210> 32
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
attgcctgct catggtgctg at 22
<210> 33
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
taacggccca attgtccttg ag 22
<210> 34
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
aggaacccag gaggatcaga gg 22
<210> 35
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
cctaccgctg ccaggaccta 20
<210> 36
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
ttgggagctg ggggaaaact 20
<210> 37
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
aaactagggc tccctcggac aa 22
<210> 38
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
aaggctgagg aatgtgggac ct 22
<210> 39
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
gggacctcca tgaccttgct t 21
<210> 40
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 40
gttgggcagg tggagtggag 20
<210> 41
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
agccattcac acgtggacca ta 22
<210> 42
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
cgtgcatgtg ttgttttctt tca 23
<210> 43
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
gctggtcagc agggattatc ag 22
<210> 44
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
tctctttagg tacaccttgt tcttctcca 29
<210> 45
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
tggtaccatc caaaatgata tactaaagga 30
<210> 46
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
gcccagattc ctgcaagtaa cg 22
<210> 47
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
aaatatagac cttgattttc tagtgtgtga tacaa 35
<210> 48
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
tacctgctca gtaagtgacc atcca 25
<210> 49
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
tgcagcagtt gctgtttaat catgt 25
<210> 50
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
tctgaaaaca aagttgcaaa gtgga 25
<210> 51
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
tcctctcaca cccccttctc ag 22
<210> 52
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
ttcaatctgc ctgtttcgca ct 22
<210> 53
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
tcaaatccag cagggatcaa gc 22
<210> 54
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
tttcttttgg gctcacatgg ttt 23
<210> 55
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
ggtcgcagac ctcgtcttct tg 22
<210> 56
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
ttgccaagtc agtggctaca tacc 24
<210> 57
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
accctcgctg ctggtcataa aa 22
<210> 58
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
atcacccctc cacacatgac aa 22
<210> 59
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 59
gtttatgctt agaaagtgga tttaatagga cctt 34
<210> 60
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 60
tctgctaaag tataggccag ctcca 25
<210> 61
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
tgctgtttat ccatttgtgt taagtggtg 29
<210> 62
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
tggcctaaga aaccttacat attacctctt 30
<210> 63
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
gggcacagag ctaatatttg gaca 24
<210> 64
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
caaaggtcag aatttttgcc aatc 24
<210> 65
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 65
taggcacttc cgggactctc ag 22
<210> 66
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 66
ttgcaaacct gcctgtacct ga 22
<210> 67
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 67
gaaaccagga agcaccagca gt 22
<210> 68
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 68
gcttgctttg tccatggagg tt 22
<210> 69
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
catggaagtt tgctacgaca ttatcc 26
<210> 70
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
ggcctgcgtt agttccagtg ac 22
<210> 71
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
tgctgttatt ttatgaagac agtgtgg 27
<210> 72
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
cctggttccc aagaaccatt ca 22
<210> 73
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
cccagccctc gtgtaaatag ga 22
<210> 74
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
gcccccattc aaagcattca ta 22
<210> 75
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
ttctctcttc ccctcaacaa gcaa 24
<210> 76
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
gtccccgact ccctctgcta ac 22
<210> 77
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
ccctctcctc tgcctcaatc ct 22
<210> 78
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
gcctatgtcc ctgggcagtt atg 23
<210> 79
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
gtggaccagc cagcaatgaa c 21
<210> 80
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
tctggcttgg cttaaacctc ctc 23
<210> 81
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
cacccatttc aggtgggatc at 22
<210> 82
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
gtgggctgtt ctgaaggcat ct 22
<210> 83
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
aacctggctt ccaggggagt ag 22
<210> 84
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
ccaatcccct ccagaaacac c 21
<210> 85
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
tccttggttc ctgtgaaggt ca 22
<210> 86
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 86
ggaatcgggg tgagatgaga ga 22
<210> 87
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 87
cccagagcta gtggctcacc tt 22
<210> 88
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 88
ctccaaatgg tgcccttcac tc 22
<210> 89
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 89
acccatggct tacatggagg tg 22
<210> 90
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 90
cgtcgctgcc aggcacaagt 20
<210> 91
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 91
ccgctgtacc tgtgtgcaca cc 22
<210> 92
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 92
ggaacagaga agggggtttg aa 22
<210> 93
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 93
catccttatt gaagggctga aatg 24
<210> 94
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 94
agatagtaag ccactttgcg gatca 25
<210> 95
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 95
ccccttgttc cacagcaact g 21
<210> 96
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 96
tgtgcttgga tgttgatctg ttga 24
<210> 97
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 97
gctgaaagag agcccagcaa tc 22
<210> 98
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 98
tgcaaatgtg ggcacagtct ct 22
<210> 99
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 99
agcacccaac atgccattac ct 22
<210> 100
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 100
tggaacaatt tggaatttga gtcgta 26
<210> 101
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 101
atacagtgcc acccacctca cc 22
<210> 102
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 102
actttgattg ttgtgtgatt tctgaagtt 29
<210> 103
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 103
ctgggctttg tgcaatgcta ac 22
<210> 104
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 104
ttttgtctgc ctgggccaag gcc 23
<210> 105
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 105
ttttcacgga cttccaggca gtgc 24
<210> 106
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 106
tgcctcagcc actcacggtt t 21
<210> 107
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 107
tatcccaacc aggttataaa tcagtg 26
<210> 108
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 108
ttttttttag accacgagat gaaaggacag 30
<210> 109
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 109
ttttttctgt gtctgtggtg ctctcatccc 30
<210> 110
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 110
tttttttttt tttcagtgac gctatgacag accac 35
<210> 111
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 111
tttttttttg gggacgccat gggtaatggt g 31
<210> 112
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 112
tttttttttt tctgttacct ttgccaagtg ttccag 36
<210> 113
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 113
ttttttttct tggtgtttta ttttcatagg cacccatt 38
<210> 114
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 114
tttttttttt ttttttgcat tgccttcctt ttgtcttag 39
<210> 115
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 115
tttttttttt tttttttttt tttttttctt cgacacgccc ccc 43
<210> 116
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 116
tttttttttt tttttttttc atgtcattta catggtccct gag 43
<210> 117
<211> 46
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 117
tttttttttt tttttttttt tttttttctg ttgcaggcac acaacc 46
<210> 118
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
tttttttttt tttttttttt ttttcggttt gctttccaga cgt 43
<210> 119
<211> 48
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
tttttttttt tttttttttt ttttttttgg aggccagcag tgaacacc 48
<210> 120
<211> 49
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
tttttttttt tttttttttt cgtttgttgc agggaggagc aagtcccca 49
<210> 121
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 121
tttttttttt tttttttttt tttttttttt tccaatcagc acagagaagg t 51
<210> 122
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 122
tttttttttt tttttttttt ttttttcact aagaaacccr gaagcactgt aatt 54
<210> 123
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 123
tttttttttt tttttttttt ttttttttca ggacgcgcag gtcatccctg tgat 54
<210> 124
<211> 55
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 124
tttttttttt tttttttttt tttttttttt tctgagggga ctcaccactt gagct 55
<210> 125
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 125
tttttttttt tttttttttt tttttttttt ttttttttgc agagcatggg tgtggac 57
<210> 126
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 126
tttttttttt tttttttttt ttttcattcc aggactgtcc attctaaatc tcatctt 57
<210> 127
<211> 59
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 127
tttttttttt tttttttttt ttttttttgt cccatttatc atcatcatgt caactttga 59
<210> 128
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 128
tttttttttt tttttttttt tttttttttt ttttttttct cccagacaga ggtgatcatc 60
c 61
<210> 129
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 129
tttttttttt tttttttttt tttttttttt tttttttttt aggcaagccc ctggagatgg 60
<210> 130
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 130
tttttttttt tttttttttt tttttttttt tttggttgtt tttggcagat aaagactgtg 60
a 61
<210> 131
<211> 65
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 131
tttttttttt tttttttttt tttttttttt gatgatgata aatgggacaa aatatccact 60
aaacc 65
<210> 132
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 132
tttttttttt tttttttttt tttttttttt tttttttttt tttttttgcc tgcccctgct 60
ccctgg 66
<210> 133
<211> 65
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 133
tttttttttt tttttttttt tttttttttt ttttttttgc acatggctct gtgctcatta 60
aagct 65
<210> 134
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 134
tttttttttt tttttttttt tttttttttt tttttttttt ttttttttta ccggttgcgc 60
ctcagccact ca 72
<210> 135
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 135
tttttttttt tttttttttt tttttttttt tttttttttt ttttttcagg accaggttga 60
tgcgtatcct tc 72
<210> 136
<211> 75
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 136
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt gagaggagcc 60
ctgtgacatt cttcg 75
<210> 137
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 137
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttttttttgt 60
gttcattgcc ctgggc 76
<210> 138
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 138
ctgtggaatt gggtgcaaa 19
<210> 139
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 139
tgctttccag acgtccatca 20
<210> 140
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 140
ttttcggtgc ccaccatgac agccgtg 27
<210> 141
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 141
tttttcaagg tctctgttcc aagttcc 27
<210> 142
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 142
tttttttttt tgccaaggcc ggggagtca 29
<210> 143
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 143
ttttttcaga gccatgtgca tttatagacc cc 32
<210> 144
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 144
tttttaagtt tgacccggaa attatcggtc ca 32
<210> 145
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 145
tttttttttt tcagacgtcc atcacggtgc tgt 33
<210> 146
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 146
ttttttttgt gtgtctaaca tagaaaggaa tctg 34
<210> 147
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 147
tttttttttt tgaagaaagt cgccatggtg ggg 33
<210> 148
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 148
tttttttttt ttttcccaga ccttctctgt gctga 35
<210> 149
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 149
tttttttttt tttactgcaa ggagtagtga gagtcaaa 38
<210> 150
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 150
tttttttttg tatctctgaa caacaccaaa atcgataaaa c 41
<210> 151
<211> 42
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 151
tttttttttt tttttttcca gagcagtggc accgcagaag tg 42
<210> 152
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 152
tttttttttt tttttttttt tcgcagaacg gcatcctcat caa 43
<210> 153
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 153
tttttttttt tttttcaaaa tcgataaaac cgattacaat ccatacc 47
<210> 154
<211> 46
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 154
tttttttttt tttttttttt ttttttacct gctgcaatgg gtatcc 46
<210> 155
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 155
tttttttttt tttttttttt ttttttttag gtgctgcctt cgcacaa 47
<210> 156
<211> 49
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 156
tttttttttt tttttttttt tgtggatatt ttgtcccatt tatcatcat 49
<210> 157
<211> 52
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 157
tttttttttt tttttttttt tttttttgac cacaacatca ttccaggact gt 52
<210> 158
<211> 56
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 158
tttttttttt tttttttttt tttttttttt ttttcacatc ccccagcagg agcacc 56
<210> 159
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 159
tttttttttt tttttttttt tttttttttt ttcctgaggg gcagtgtggc caca 54
<210> 160
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 160
tttttttttt tttttttttt ttttttttat ctctgaacaa caccaaaatc gataaaa 57
<210> 161
<211> 56
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 161
tttttttttt tttttttttt ttttttttga tctcccaact ggaaggggtg cagcaa 56
<210> 162
<211> 59
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 162
tttttttttt tttttttttt tttttttttt tttgctctgt gctcattaaa gctacccac 59
<210> 163
<211> 59
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 163
tttttttttt tttttttttt tttttttttt ttttgcctga acgtagaagt accacccac 59
<210> 164
<211> 62
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 164
tttttttttt tttttttttt tttttttttt tttccacgct cactctgata aggacgacgt 60
cg 62
<210> 165
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 165
tttttttttt tttttttttt tttttttttt ttttggtgag gacaatttaa tcatcaggtg 60
a 61
<210> 166
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 166
tttttttttt tttttttttt tttttttttt tttttttttt ttttttagtt caacatgggc 60
gttcatctct ta 72
<210> 167
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 167
tttttttttt tttttttttt tttttttttt ttttttgttg gctgacctgt gtctcagaga 60
ttt 63
<210> 168
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 168
tttttttttt tttttttttt tttttttttt tttttttttt tctgtgatct gtcccactcc 60
ctt 63
<210> 169
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 169
tttttttttt tttttttttt tttttttttt tttttttttt ttttttttgc caagtgttcc 60
agccac 66
<210> 170
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 170
tttttttttt tttttttttt tttttttttt tttttttttt tctgtgggat tcttgccatc 60
ctgtgt 66
<210> 171
<211> 69
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 171
tttttttttt tttttttttt tttttttttt tttttttttt tttttgcatg aaggtttccc 60
tggaacaag 69
<210> 172
<211> 75
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 172
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttagctcag 60
cgtgtaacgt gcgct 75
<210> 173
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 173
tttttttttt ttttcagcag gagggataaa ctaacc 36
<210> 174
<211> 48
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 174
tttttttttt tttttttttt tttttgcttc gaggtttacc ccttccag 48
<210> 175
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 175
tttttttttt tttttttttt tttttttttt ttttttttag gcgtgctgag taccagg 57
<210> 176
<211> 77
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 176
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttcac 60
tgggtcagtc agcattc 77
<210> 177
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 177
tttttttttt ctaccatctg ggccgtgtta 30
<210> 178
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 178
tttttttttt tttttttttt tttttttgca gatagaaatc attgctgact a 51
<210> 179
<211> 78
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 179
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttgac 60
tacgaggtrc accccaac 78
<210> 180
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 180
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt taaccgaygc 60
cccaagatcc 70
<210> 181
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 181
tttttttttt tttttttttt tttttggtag tagtaagcag ccccagctac a 51
<210> 182
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 182
ttttttttgg tcagcctcca catcttgtc 29
<210> 183
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 183
tttttttttt tttttttttt tttccccaaa tcgggggtgt atgc 44
<210> 184
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 184
tttttttttt tttttttttt tttttttttt ttttttttta gcgcatatca ggtgtgtgca 60
tac 63
<210> 185
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 185
tttttttttt tttggcagca gcactacccc t 31
<210> 186
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 186
tttttttttt tttttttttt tttttttttt tttttttttg tacctacaag agctgaccgt 60
<210> 187
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 187
tttttttttt tttttttttg tgtcacagca tcccggtaag t 41
<210> 188
<211> 56
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 188
tttttttttt tttttttttt ttttttttca atttccactg gcgtgattgg acttac 56
<210> 189
<211> 46
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 189
tttttttttt tttttttttt tttttagtag cgctcctggg gtgtca 46
<210> 190
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 190
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tgtggagtgg 60
agaagtaggc ct 72
<210> 191
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 191
tttttttttt tttttttttt tttttcacag tactaaaacc agcatactgg 50
<210> 192
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 192
tttttttttc tgaatcatag ccacgatgtg 30
<210> 193
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 193
ttttgaatgg ctggagttgc tattg 25
<210> 194
<211> 62
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 194
tttttttttt tttttttttt tttttttttt ttttttaatt cctttagaaa aaatgtcagt 60
tt 62
<210> 195
<211> 69
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 195
tttttttttt tttttttttt tttttttttt tttttttttt tgcagttatt ccagttcttg 60
caaatttta 69
<210> 196
<211> 62
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 196
tttttttttt tttttttttt tttttttttt tttttgttct ggaggaaaaa tgtatgtatt 60
tc 62
<210> 197
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 197
tttttttttt tttttttttt tttttttttt ttttcctttg ctgtatattc aaatatgtca 60
<210> 198
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 198
tttttttttt tttttttttt tttttttttt tttttttctt attcactgtt acaatttttt 60
tttttatgta 70
<210> 199
<211> 56
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 199
tttttttttt tttttttttt tttttttttt tgaaattgaa ttaaattttg gcatgt 56
<210> 200
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 200
tttttttttt tttttttttt tttttttttt tttttttgct tcctgcatat ggtatcc 57
<210> 201
<211> 67
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 201
tttttttttt tttttttttt tttttttttt tttttagaaa ttcattccaa ttccccagaa 60
gaaagac 67
<210> 202
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 202
tttttttttt tttttttttt tttttttgca aagatgagag ctggtagaag a 51
<210> 203
<211> 71
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 203
tttttttttt tttttttttt tttttttttt tttttttttt tttcttcttc taagagcaca 60
ctgtcagaca t 71
<210> 204
<211> 49
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 204
tttttttttt tttttttttt ttttttcttt gagtttttgt tttttatca 49
<210> 205
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 205
tttgccattg cttctattgc agta 24
<210> 206
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 206
tttttttttt tttttttttg tgagacattc catcatgyaa gaa 43
<210> 207
<211> 52
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 207
tttttttttt tttttttttt ttttttgtct cacaaatgat gtttatgatg ct 52
<210> 208
<211> 49
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 208
tttttttttt tttttttttt ttttaaaatt tttctttggg aaattacca 49
<210> 209
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 209
tttttttttc aagagccagg atatttccat ct 32
<210> 210
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 210
tttttttttt ttttcatctg tgatttctcc cactgtacat 40
<210> 211
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 211
tttttttttt ttcgatatgc tccactcacc attc 34
<210> 212
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 212
aaaatgggac aagatggcc 19
<210> 213
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 213
tttttttttt agtttttagt acctggaaaa gaggg 35
<210> 214
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 214
tttttttttt tttttttttt tttttttttt tccctagttc ctgaactcat caaa 54
<210> 215
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 215
tttttttttt tttttttttt ttacacatga caagaatatc tggcc 45
<210> 216
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 216
tttttttttt ttttttttca tcccagcttg cactggtttc c 41
<210> 217
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 217
ttttttacag gaaattggcc ttgcct 26
<210> 218
<211> 69
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 218
tttttttttt tttttttttt tttttttttt tttttttttt ttttttttta ctgtgtgcag 60
tggaagact 69
<210> 219
<211> 68
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 219
tttttttttt tttttttttt tttttttttt tttttttttt tttttcttgt ataaaaccca 60
tgcttgct 68
<210> 220
<211> 77
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 220
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttatc 60
ctgtgtaccg tgcaaga 77
<210> 221
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 221
ttttttttgg cccttctcag ttcgctac 28
<210> 222
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 222
tttttcccct caacaagcaa ggcagactta 30
<210> 223
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 223
tttttttgta ctgtgtgcag tggaagactc 30
<210> 224
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 224
ttttcttagg aactttgctg ccacaatacc t 31
<210> 225
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 225
tttctttgag ttttctttct tcttttcatc cca 33
<210> 226
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 226
tttttttctg trgctggcct gctttcctct 30
<210> 227
<211> 77
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 227
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttttcggagt 60
tccagcccct stattac 77
<210> 228
<211> 79
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 228
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttagctg 60
gaggacagta ctcacggtt 79
<210> 229
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 229
tttttttttt tttttttgtg tcccgccgca tctcc 35
<210> 230
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 230
tttttttttt tctctgtgct ttctgtcttt cagtg 35
<210> 231
<211> 79
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 231
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttggaga 60
ctccataaag gcatatggt 79
<210> 232
<211> 74
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 232
tttttttttt tttttttttt tttttttttt tttttttttt tttttttaaa ccacacaaat 60
attttctgca cata 74
<210> 233
<211> 85
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 233
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt 60
aactggtagc tggaggacag tactc 85
<210> 234
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 234
tttttttttt tttttttttt cccaggcttg ggcaggaaac tct 43
<210> 235
<211> 75
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 235
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tcgcatctcc 60
ttcagcgcga gccac 75
<210> 236
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 236
tttttttttt tttttttgag ttttaatgat gccaaggaga aagt 44
<210> 237
<211> 83
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 237
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt 60
taactggatg ctgaccaccc tgt 83
<210> 238
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 238
tttttttttt tttttttttt tccgagtggc ctcgtaaggt gtaaatt 47
<210> 239
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 239
tttttttttt tttttttttt tttttttgga actttgctgc cacaatacct c 51
<210> 240
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 240
tttttttttt tttttttttt ttttttttca tggatccaag cccatgtata 50
<210> 241
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 241
tttttttttt tttttttttt ttttttagtg gcctcgtaag gtgtaaatta c 51
<210> 242
<211> 53
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 242
tttttttttt tttttttttt tttttttttt ttcttcctcc atgtattcca ctc 53
<210> 243
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 243
tttttttttt tttttttttt ttttgcaaag tatggcacat ccatatccag attc 54
<210> 244
<211> 55
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 244
tttttttttt tttttttttt cctctaattc ttacctgtgt ctttcttctt atctc 55
<210> 245
<211> 56
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 245
tttttttttt tttttttttt tttttttttt ttttttttca cttggggcct acagta 56
<210> 246
<211> 58
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 246
tttttttttt tttttttttt ttttgtaagg tgtaaattac ttactgttaa tggaatca 58
<210> 247
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 247
tttttttttt tttttttttt tttttttttt ttttttttgg cagacttact ggcggtagtt 60
<210> 248
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 248
tttttttttt tttttttttt tttttttttt tttttgccta cagtactgct tatcagagaa 60
g 61
<210> 249
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 249
tttttttttt tttttttttt tttttttttt tttttttttt cctctcggga tttcttgggt 60
g 61
<210> 250
<211> 65
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 250
tttttttttt tttttttttt tttttttttt tttttttttc tagggtttta aagatcctgt 60
gtacc 65
<210> 251
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 251
tttttttttt tttttttttt tttttttttt tttttttttt ttttctgctt tcctctcggg 60
atttct 66
<210> 252
<211> 62
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 252
tttttttttt tttttttttt tttttttttt tttttttttt tttactggat gctgaccacc 60
ct 62
<210> 253
<211> 89
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 253
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt 60
ttttttgcaa acaaggagac tccataaag 89
<210> 254
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 254
cttgcactgg tttccgcctc 20
<210> 255
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 255
ttcatcccag cttgcactgg tt 22
<210> 256
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 256
tttaaaaatc aggtgtctct tttctccta 29
<210> 257
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 257
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt cccagcttgc 60
actggtttcc gc 72
<210> 258
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 258
tttttttttt tagccagatt cataaccatt ccc 33
<210> 259
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 259
tttttttttt tgcaatgtca gcaaactgct tc 32
<210> 260
<211> 83
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 260
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt 60
tttctccttg tttgcagagc cca 83
<210> 261
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 261
ttttttggta gctggaggac agtactcacg gt 32
<210> 262
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 262
tttttttccc agatataaac agctacattt tcagtc 36
<210> 263
<211> 80
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 263
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttttttttgg 60
tagctggagg acagtactca 80
<210> 264
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 264
tttttttttt tttttgtctt tcagtgacat ctgccat 37
<210> 265
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 265
tttttttttt ttttaaatgg tgccatatca ctgatct 37
<210> 266
<211> 80
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 266
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttttcccttg 60
atcataagaa tctggatatg 80
<210> 267
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 267
tttttttttt tttttttgta ttgtccaaga cctcaatcct t 41
<210> 268
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 268
tttttttttt ttttttttac ctgacccaca ttgaatctag acct 44
<210> 269
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 269
tttttttttt tttttttttt ttctcatcct acgggccatg gactc 45
<210> 270
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 270
tttttttttt tttttttttt tttgagccca aactccacag taaacca 47
<210> 271
<211> 86
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 271
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttttttttg 60
ctctcagatt gactttccat tccaga 86
<210> 272
<211> 49
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 272
tttttttttt tttttttttt cattatccaa gacaarcatg attgtagca 49
<210> 273
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 273
tttttttttt tttttttttt tttttttttt tagacatgga tccaagccca 50
<210> 274
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 274
tttttttttt tttttttttt ttttttggtt ttaaagatcc tgtgtaccgt 50
<210> 275
<211> 51
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 275
tttttttttt tttttttttt tttttggcga gcttttcaat gtattcatca g 51
<210> 276
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 276
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttttttcagt 60
tcgctacgac ccatac 76
<210> 277
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 277
tttttttttt tttttttttt ttttttccaa gacaarcatg attgtagcac tgac 54
<210> 278
<211> 70
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 278
tttttttttt tttttttttt tttttttttt tttttttttt tttagctttt caatgtattc 60
atcaggtgca 70
<210> 279
<211> 58
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 279
tttttttttt tttttttttt tttttttttt ttttttaatc caaatggcca tgggcaca 58
<210> 280
<211> 73
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 280
tttttttttt tttttttttt tttttttttt tttttttttt ttttaaaaaa tccattcctt 60
acctgggaaa act 73
<210> 281
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 281
tttttttttt tttttttttt tttttttttc ctccagaaag ttcttcctgt aggagtt 57
<210> 282
<211> 59
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 282
tttttttttt tttttttttt tttttttctg tgtctttctt cttatctcgt gaaagctca 59
<210> 283
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 283
tttttttttt tttttttttt tttttttttt tttttttttt ttgcactggt ttccgcctcc 60
<210> 284
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 284
tttttttttt tttttttttt tttttttttt tttatgattg tagcactgac ctcaaataag 60
<210> 285
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 285
tttttttttt tttttttttt tttttttttt ttttttttta tacctctcca tgtactgtca 60
cca 63
<210> 286
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 286
tttttttttt tttttttttt tttttttttt tttttttttt ttttttcacc tttgtcacca 60
cctcac 66
<210> 287
<211> 68
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 287
tttttttttt tttttttttt tttttttttt tttttttttt tttcaarcat gattgtagca 60
ctgacctc 68
<210> 288
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 288
tttttttttt tttttttttc acttggggcc tacagtact 39
<210> 289
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 289
cagctccagg gggagaagct tt 22
<210> 290
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 290
ttggtagttg taggcaatgt cagca 25
<210> 291
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 291
tttttttttg gccaggccac ccaagaaatc c 31
<210> 292
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 292
ttttttagat tgactttcca ttccagattt act 33
<210> 293
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 293
tttttttttt tcccagctgg aagacgtttc t 31
<210> 294
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 294
tttttttttt ttttaacagg ttctattttc cccca 35
<210> 295
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 295
tttttttttt ttctgataag cagtactgta ggccc 35
<210> 296
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 296
ttttttttgg agaatgatgt aaacctgacc caca 34
<210> 297
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 297
tttttttttt tttggtagct ggaggacagt actcac 36
<210> 298
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 298
ttttttttta agtactgtgg cttccatgaa gataaca 37
<210> 299
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 299
ttttttttta cttttcaaga agtggaaaaa tgtgattg 38
<210> 300
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 300
tttttttttt tttttttttt taaatcccga gaggaaagca ggc 43
<210> 301
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 301
tttttttttt tttttttttt cgatctgaaa acaagggcac atgtc 45
<210> 302
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 302
tttttttttt tttttttttt aatcctttgg gtgtatgggt cgtag 45
<210> 303
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 303
tttttttttt tttttttttt tttgcctgcc tgctctgaca aacrtca 47
<210> 304
<211> 49
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 304
tttttttttt tttttttttt tcaccttact ttctccttgg catcattaa 49
<210> 305
<211> 52
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 305
tttttttttt tttttttttt ttttttttta tcaggtgcac ccagagaggc aa 52
<210> 306
<211> 52
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 306
tttttttttt tttttttttt tttttttttt atctgctcat gggccttcca ct 52
<210> 307
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 307
tttttttttt tttttttttt tttttttggc taaaaaatcc attccttacc tggg 54
<210> 308
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 308
tttttttttt tttttttttt tttttttttt gaagataaca ttccccagct ggaagac 57
<210> 309
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 309
tttttttttt tttttttttt tttttttttc tcctagggtt ttaaagatcc tgtg 54
<210> 310
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 310
tttttttttt tttttttttt tttttttttt tgacagccat csaaaattac actgtca 57
<210> 311
<211> 58
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 311
tttttttttt tttttttttt tttttttttt ttttttgaaa tcccgagagg aaagcagg 58
<210> 312
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 312
tttttttttt tttttttttt tttttttcta ttcatagcat gggaaatcta agaggtataa 60
<210> 313
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 313
tttttttttt tttttttttt tttttttttt ttggtcatac ctgtaattca ccaaaggatg 60
<210> 314
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 314
tttttttttt tttttttttt tttttttttt ttttttcaga cttactggcg gtagttgtag 60
<210> 315
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 315
tttttttttt tttttttttt tttttttttt tttttttttt ttaagatcct gtgtaccgtg 60
caa 63
<210> 316
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 316
tttttttttt tttttttttt tttttttttt tttttttttt ttccatttgg attgttgcat 60
ttccca 66
<210> 317
<211> 67
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 317
tttttttttt tttttttttt tttttttttt tttttttttt tttttcatcc cagcttgcac 60
tggtttc 67
<210> 318
<211> 71
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 318
tttttttttt tttttttttt tttttttttt tttttttttt ttttcctgtg gttttggtct 60
taggaacttt g 71
<210> 319
<211> 74
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 319
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttc ttcttttcat 60
cccagcttgc actg 74
<210> 320
<211> 73
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 320
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt gtctctagga 60
ggcgattatg cag 73
<210> 321
<211> 76
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 321
tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt ttttgcggaa 60
ctggatgctg accacc 76

Claims (14)

1.一种基因突变检测试剂盒，其特征在于，所述试剂盒包含特异性检测ATP7B基因的突变位点的核酸检测试剂；并且，所述ATP7B基因的突变位点包括下组A的突变位点：
组A
所述试剂盒包含针对组A的ATP7B基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:1-32所示；所述单碱基延伸引物的序列如SEQ ID NO:104-136所示。
2.如权利要求1所述的试剂盒，其特征在于，所述ATP7B基因的突变位点还包括选自下组B的突变位点：
组B
所述试剂盒包含针对组B的突变位点的单碱基延伸引物，所述单碱基延伸引物的序列如SEQ ID NO:137-171所示。
3.如权利要求1所述的试剂盒，其特征在于，所述试剂盒还包含特异性检测MMACHC基因的突变位点的核酸检测试剂，并且，所述MMACHC基因的突变位点选自下组C：
组C
所述试剂盒包含针对组C的MMACHC基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:33-40所示；所述单碱基延伸引物的序列如SEQ ID NO:172-190所示。
4.如权利要求1所述的试剂盒，其特征在于，所述试剂盒还包含特异性检测MUT基因的突变位点的核酸检测试剂，并且，所述MUT基因的突变位点选自下组D：
组D
突变位点 染色体上位置信息(GRCh38) MUT:c.323G>A 6:49459144 MUT:c.424A>G 6:49458020 MUT:c.494A>G 6:49457950 MUT:c.554C>T 6:49457890 MUT:c.599T>C 6:49457845 MUT:c.682C>T 6:49457762 MUT:c.729_730insTT 6:49457714-49457715 MUT:c.754-1G>A 6:49456238 MUT:c.755dupA 6:49456235-49456236 MUT:c.786T>G 6:49456205 MUT:c.914T>C 6:49453754 MUT:c.982C>T 6:49453686 MUT:c.1061C>T 6:49453607 MUT:c.1084-10A>G 6:49451724 MUT:c.1106G>A 6:49451692 MUT:c.1280G>A 6:49451518 MUT:c.1323_1324insA 6:49451474-49451475 MUT:c.1445-1G>A 6:49447786 MUT:c.1630_1631delGGinsTA 6:49444684-49444685 MUT:c.1677-1G>A 6:49441972 MUT:c.1741C>T 6:49441907 MUT:c.1880A>G 6:49440282 MUT:c.1943G>A 6:49440219 MUT:c.2062G>T 6:49435518 MUT:c.2080C>T 6:49435500
所述试剂盒包含针对组D的MUT基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:41-62所示；所述单碱基延伸引物的序列如SEQ ID NO:191-215所示。
5.如权利要求1所述的试剂盒，其特征在于，所述试剂盒还包含特异性检测PAH基因和PTS基因的突变位点的核酸检测试剂，并且，所述PAH基因和PTS基因的突变位点选自下组E：
组E
所述试剂盒包含针对组E的基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:63-103所示；所述单碱基延伸引物的序列如SEQ ID NO:216-251所示。
6.如权利要求1所述的试剂盒，其特征在于，所述试剂盒还包含特异性检测PAH基因和PTS基因的突变位点的核酸检测试剂，并且，所述PAH基因和PTS基因的突变位点选自下组F：
组F
所述试剂盒包含针对组F的基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:63-103所示；所述单碱基延伸引物的序列如SEQ ID NO:252-288所示。
7.如权利要求1所述的试剂盒，其特征在于，所述试剂盒还包含特异性检测PAH基因和PTS基因的突变位点的核酸检测试剂，并且，所述PAH基因和PTS基因的突变位点选自下组G：
组G
所述试剂盒包含针对组G的基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:63-103所示；所述单碱基延伸引物的序列如SEQ ID NO:289-321所示。
8.如权利要求1-7中任一项所述的试剂盒，其特征在于，所述试剂盒包含如SEQ ID NO:1-103所示的多重PCR引物和以下的单碱基延伸引物：
针对组A的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:104-136所示，
针对组B的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:137-171所示，
针对组C的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:172-190所示，
针对组D的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:191-215所示，
针对组E的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:216-251所示，
针对组F的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:252-288所示，和
针对组G的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:289-321所示。
9.一种核酸检测试剂在制备基因突变检测试剂盒中的用途，所述ATP7B基因的突变位点包括组A的突变位点；
组A
所述的核酸检测试剂包括多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:1-32所示；所述单碱基延伸引物的序列如SEQ ID NO:104-136所示。
10.如权利要求9所述的用途，其特征在于，所述ATP7B基因的突变位点还包括选自组B的突变位点；
组B
并且所述的核酸检测试剂进一步包括针对组B的突变位点的单碱基延伸引物，所述单碱基延伸引物的序列如SEQ ID NO:137-171所示。
11.如权利要求9所述的用途，其特征在于，所述的核酸检测试剂还包括特异性检测MMACHC基因的突变位点的核酸检测试剂，并且，所述MMACHC基因的突变位点选自组C；
组C
并且所述的核酸检测试剂进一步包括针对组C的MMACHC基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:33-40所示；所述单碱基延伸引物的序列如SEQ ID NO:172-190所示。
12.如权利要求9所述的用途，其特征在于，所述的核酸检测试剂包括特异性检测MUT基因的突变位点的核酸检测试剂，并且，所述MUT基因的突变位点选自组D；
组D
并且所述的核酸检测试剂进一步包括针对组D的MUT基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:41-62所示；所述单碱基延伸引物的序列如SEQ ID NO:191-215所示。
13.如权利要求9所述的用途，其特征在于，所述的核酸检测试剂包括特异性检测PAH基因和PTS基因的突变位点的核酸检测试剂，并且，所述PAH基因和PTS基因的突变位点选自组E、组F和组G；
组E
组F
组G
并且所述的核酸检测试剂进一步包括针对组E、组F和组G的基因突变位点的多重PCR引物和单碱基延伸引物，所述多重PCR引物的序列如SEQ ID NO:63-103所示；所述单碱基延伸引物的序列选自如SEQ ID NO:216-321所示。
14.如权利要求9所述的用途，其特征在于，所述基因突变检测试剂盒用于检测如组A-组G所示的ATP7B、PAH、PTS、MUT和MMACHC基因的突变，
组B
组C
组D
突变位点 染色体上位置信息(GRCh38) MUT:c.323G>A 6:49459144 MUT:c.424A>G 6:49458020 MUT:c.494A>G 6:49457950 MUT:c.554C>T 6:49457890 MUT:c.599T>C 6:49457845 MUT:c.682C>T 6:49457762 MUT:c.729_730insTT 6:49457714-49457715 MUT:c.754-1G>A 6:49456238 MUT:c.755dupA 6:49456235-49456236 MUT:c.786T>G 6:49456205 MUT:c.914T>C 6:49453754 MUT:c.982C>T 6:49453686 MUT:c.1061C>T 6:49453607 MUT:c.1084-10A>G 6:49451724 MUT:c.1106G>A 6:49451692 MUT:c.1280G>A 6:49451518 MUT:c.1323_1324insA 6:49451474-49451475 MUT:c.1445-1G>A 6:49447786 MUT:c.1630_1631delGGinsTA 6:49444684-49444685 MUT:c.1677-1G>A 6:49441972 MUT:c.1741C>T 6:49441907 MUT:c.1880A>G 6:49440282 MUT:c.1943G>A 6:49440219 MUT:c.2062G>T 6:49435518 MUT:c.2080C>T 6:49435500
组E
组F
组G
突变位点 染色体上位置信息(GRCh38) PAH:c.1085C>T 12:102843760 PAH:c.482T>C 12:102866623 PAH:c.755G>A 12:102852902 PAH:c.977G>A 12:102844424 PAH:c.694C>T 12:102855148 PAH:c.913-7A>G 12:102846958 PAH:c.1066-11G>A 12:102843790 PAH:c.194T>C 12:102894893 PAH:c.842C>T 12:102852815 PAH:c.671T>C 12:102855171 PAH:c.617A>G 12:102855225 PAH:c.739G>C 12:102852918 PAH:c.865G>C 12:102851734 PAH:c.1238G>C 12:102840477 PAH:c.284_286delTCA 12:102894801-102894803 PAH:c.1174T>A 12:102843671 PAH:c.922C>T 12:102846942 PTS:c.84-291A>G 11:112228303 PAH:c.907T>C 12:102851692 PAH:c.688G>A 12:102855154 PAH:c.460T>C 12:102866645 PAH:c.1139C>T 12:102843706 PAH:c.740G>T 12:102852917 PAH:Ex1del3758 12:102917536-102921293 PAH:c.1045T>G 12:102844356 PAH:c.494C>A 12:102866611 PAH:c.470G>T/A 12:102866635 PTS:c.118_121delTTTG 11:112228628-112228631 PAH:c.721C>T 12:102852936 PAH:c.1208C>T 12:102840507 PAH:c.716G>A 12:102852941 PTS:c.259C>T 11:112233178 PAH:c.440C>T 12:102877463
所述的核酸检测试剂包括如SEQ ID NO:1-103所示的多重PCR引物和以下的单碱基延伸引物：
针对组A的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:104-136所示，
针对组B的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:137-171所示，
针对组C的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:172-190所示，
针对组D的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:191-215所示，
针对组E的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:216-251所示，
针对组F的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:252-288所示，和
针对组G的突变位点的单碱基延伸引物，并且所述单碱基延伸引物的序列如SEQ IDNO:289-321所示。
CN202010754298.0A 2020-07-30 2020-07-30 一种基因突变检测试剂盒及其应用 Active CN114058689B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010754298.0A CN114058689B (zh) 2020-07-30 2020-07-30 一种基因突变检测试剂盒及其应用

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010754298.0A CN114058689B (zh) 2020-07-30 2020-07-30 一种基因突变检测试剂盒及其应用

Publications (2)

Publication Number Publication Date
CN114058689A CN114058689A (zh) 2022-02-18
CN114058689B true CN114058689B (zh) 2024-08-20

Family

ID=80227284

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010754298.0A Active CN114058689B (zh) 2020-07-30 2020-07-30 一种基因突变检测试剂盒及其应用

Country Status (1)

Country Link
CN (1) CN114058689B (zh)

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017207797A1 (en) * 2016-06-03 2017-12-07 Inserm (Institut National De La Sante Et De La Recherche Medicale) Diet controlled expression of a nucleic acid encoding cas9 nuclease and uses thereof
CN119028454A (zh) * 2017-06-19 2024-11-26 美国控股实验室公司 通过综合计算和实验深度突变学习框架解释基因和基因组变体
JP7588578B2 (ja) * 2018-07-09 2024-11-22 フラッグシップ パイオニアリング イノベーションズ ブイ， インコーポレイテッド フソソームの組成物及びその使用
WO2020076892A1 (en) * 2018-10-09 2020-04-16 The University Of North Carolina At Chapel Hill Regulated gene editing system
CN110499364A (zh) * 2019-07-30 2019-11-26 北京凯昂医学诊断技术有限公司 一种用于检测扩展型遗传病全外显子的探针组及其试剂盒和应用
CN110468194A (zh) * 2019-08-12 2019-11-19 广州万德基因医学科技有限公司 用于遗传代谢病高通量测序建库的多重pcr引物组及试剂盒
CN111455041A (zh) * 2020-05-07 2020-07-28 和卓生物科技（上海）有限公司 一种用于基因筛查的探针组合及其应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
"中国人WD患者基因突变谱的研究及其基因型与中医证型的关系分析";张海云;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》(第12期);第18-23页 *

Also Published As

Publication number Publication date
CN114058689A (zh) 2022-02-18

Similar Documents

Publication Publication Date Title
CN109402257B (zh) 一种用于人类brca1和brca2基因全编码序列突变位点检测的引物、方法和试剂盒
JP2008538496A (ja) ウルトラディープ配列決定を用いて配列変異体を決定するための方法
CN111235272B (zh) 一次性检测肺癌多重基因突变的组合物及其应用
CN107058538B (zh) 一种引物组合物及其组成的试剂盒和应用
Hahn et al. Recent progress in non-invasive prenatal diagnosis
KR101446556B1 (ko) Ｍｔｈｆｒ 유전자 증폭용 프라이머 세트, 그것을 포함하는 ｍｔｈｆｒ 유전자 증폭용 시약 및 그 용도
CN106367491A (zh) 检测耳聋易感基因的试剂盒
CN110564838A (zh) 用于新生儿糖原累积病基因分型的多重pcr引物系统及其用途
CN112941635A (zh) 一种提高文库转化率的二代测序建库试剂盒及其方法
CN116445608B (zh) 检测耳聋相关基因突变的组合物、试剂盒、及用途
CN105969879B (zh) 一种高通量检测AhFAD2A基因突变位点分型的引物组及检测方法
CN104450869A (zh) 一种双脱氧核苷修饰的引物方法、反应体系及其在突变检测中的应用
CN111500748B (zh) 用于检测617种SNP和InDel的引物组合及其在法医鉴定和亲缘关系鉴定中的应用
WO2022007863A1 (zh) 一种靶基因区域快速富集方法
CN114058689B (zh) 一种基因突变检测试剂盒及其应用
CN114774409A (zh) 基于224个InDel和57个SNP位点的二代测序检测体系
CN118308502A (zh) 检测结核分枝杆菌耐药基因的引物组、试剂盒及方法
KR102087024B1 (ko) 배추의 순도 검정 및 조기 고정 계통 선발을 위한 단일염기다형성 마커 세트 및 이의 용도
CN115927356B (zh) Slc45a2致病突变基因、致病突变体及在制备眼皮肤白化病ⅳ型诊断试剂盒中的应用
CN108841929B (zh) 一种人ATP7b基因外显子PCR扩增系统、检测试剂盒及其应用的方法
CN109750098B (zh) Atp7b基因大片段缺失检测试剂盒及检测方法
CN112662748B (zh) 用于单细胞dna测序文库质量鉴定的引物对组合、方法和试剂盒
US7049070B2 (en) Materials and methods for detection and characterization of nucleic acid sequence variability
CN109837274B (zh) 一种人线粒体基因组文库构建的方法及应用
CN114908159A (zh) 结直肠进展期腺瘤的筛查、风险评估及预后方法和试剂盒

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant