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

CN114787347A - 新颖的mad核酸酶 - Google Patents

新颖的mad核酸酶 Download PDF

Info

Publication number
CN114787347A
CN114787347A CN202080086100.4A CN202080086100A CN114787347A CN 114787347 A CN114787347 A CN 114787347A CN 202080086100 A CN202080086100 A CN 202080086100A CN 114787347 A CN114787347 A CN 114787347A
Authority
CN
China
Prior art keywords
lys
leu
glu
ile
ser
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.)
Granted
Application number
CN202080086100.4A
Other languages
English (en)
Other versions
CN114787347B (zh
Inventor
本杰明·米杰茨
金主汉
阿米尔·米尔
安德鲁·加斯特
凯尔·西蒙
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.)
Inscripta Inc
Original Assignee
Inscripta Inc
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 Inscripta Inc filed Critical Inscripta Inc
Priority to CN202410829829.6A priority Critical patent/CN118638870A/zh
Publication of CN114787347A publication Critical patent/CN114787347A/zh
Application granted granted Critical
Publication of CN114787347B publication Critical patent/CN114787347B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases RNAses, DNAses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111—General methods applicable to biologically active non-coding nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPRs]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

本公开内容提供了用于对活细胞中的核酸进行合理、直接的编辑的新的RNA引导的核酸酶。本公开内容提供了在哺乳动物细胞中具有不同的PAM偏好和/或不同的活性的挖掘的MAD系列核酸酶(例如RNA引导的核酸酶或RGN)。在一些方面，MAD系列系统组分作为待转录的序列(在gRNA组分的情况下)和待转录和翻译的序列(在MAD系列核酸酶的情况下)被递送，并且在一些方面，MAD系列核酸酶的编码序列和gRNA组分序列在同一载体上。

Description

新颖的MAD核酸酶
相关案例
本国际PCT申请要求于2019年12月10日提交的题为“Novel MAD Nucleases”的USSN 62/946,282的优先权。
发明领域
本发明涉及用于对活细胞进行合理、直接的编辑的新颖的核酸引导的核酸酶。
发明背景
在以下讨论中，将出于背景和介绍的目的描述某些文章和方法。本文包含的任何内容不被解释为对现有技术的“承认”。本申请人明确地保留根据情况证明根据可适用的法定条文由本文引用的方法不构成现有技术的权利。
对活细胞的基因组进行精确的、靶向的改变的能力一直是生物医学研究和开发的长期目标。最近，已经鉴定出允许操纵基因序列从而操纵基因功能的多种核酸酶。这些核酸酶包括核酸引导的核酸酶。然而，核酸引导的核酸酶可以识别的靶序列的范围受需要特定PAM定位于期望的靶序列附近的限制。PAM是由gRNA/核酸酶复合体识别的短核苷酸序列，在该处gRNA/核酸酶复合体指导靶序列的编辑。用于不同的核酸引导的核酸酶的精确PAM序列和PAM长度要求不同；然而，PAM通常是与靶序列邻近或接近的2-7个碱基对序列，并且取决于核酸酶，可以在靶序列的5’或3’。核酸引导的核酸酶的工程化或挖掘新的核酸引导的核酸酶可提供具有改变的PAM偏好和/或改变的活性或保真度的核酸酶；可以增加核酸引导的核酸酶对某些编辑任务的多功能性(versatility)的所有改变。
因此，在核酸引导的核酸酶基因编辑领域中存在对具有不同的PAM偏好、在来自不同生物体诸如哺乳动物的细胞中的不同活性和/或改变的酶保真度的新颖的核酸酶的需求。本文描述的新颖的MAD系列核酸酶满足了这一需求。
发明概述
提供该概述是为了以简化的形式介绍概念的选择，所述概念在下文的详述中进一步描述。该概述既不意图确定所要求保护的主题的关键或基本特征，也不意图用于限制所要求保护的主题的范围。所要求保护的主题的其他特征、细节、效用和优势将是从下面撰写的详述，包括附图中阐明的和所附的权利要求中限定的那些方面明显的。
本公开内容提供了哺乳动物细胞中的具有不同PAM偏好和/或不同活性的挖掘的MAD系列核酸酶(例如，RNA引导的核酸酶或RGN)。
因此，在一种实施方案中，提供了执行核酸引导的核酸酶编辑的MAD系列核酸酶系统，包括以下系统：包括SEQ ID No.1(MAD2001核酸酶)、SEQ ID No.20(gRNA重复序列)和SEQ ID No.21(gRNA tracr序列)的MAD2001系统；包括SEQ ID No.1(MAD2001核酸酶)、SEQID No.22(gRNA重复序列)和SEQ ID No.23(gRNA tracr序列)的MAD2001系统；包括SEQ IDNo.1(MAD2001核酸酶)、SEQ ID No.24(gRNA重复序列)和SEQ ID No.25(gRNA tracr序列)的MAD2001系统；包括SEQ ID No.7(MAD2007核酸酶)、SEQ ID No.26(gRNA重复序列)和SEQID No.27(gRNA tracr序列)的MAD2007系统；包括SEQ ID No.7(MAD2007核酸酶)、SEQ IDNo.28(gRNA重复序列)和SEQ ID No.29(gRNA tracr序列)的MAD2007系统；包括SEQ IDNo.7(MAD2007核酸酶)、SEQ ID No.30(gRNA重复序列)和SEQ ID No.31(gRNA tracr序列)的MAD2007系统；包括SEQ ID No.8(MAD2008核酸酶)、SEQ ID No.32(gRNA重复序列)和SEQID No.33(gRNA tracr序列)的MAD2008系统；包括SEQ ID No.8(MAD2008核酸酶)、SEQ IDNo.34(gRNA重复序列)和SEQ ID No.35(gRNA tracr序列)的MAD2008系统；包括SEQ IDNo.8(MAD2008核酸酶)、SEQ ID No.36(gRNA重复序列)和SEQ ID No.37(gRNA tracr序列)的MAD2008系统；包括SEQ ID No.9(MAD2009核酸酶)、SEQ ID No.38(gRNA重复序列)和SEQID No.39(gRNA tracr序列)的MAD2009系统；包括SEQ ID No.9(MAD2009核酸酶)、SEQ IDNo.38(gRNA重复序列)和SEQ ID No.40(gRNA tracr序列)的MAD2009系统；包括SEQ IDNo.9(MAD2009核酸酶)、SEQ ID No.41(gRNA重复序列)和SEQ ID No.42(gRNA tracr序列)的MAD2009系统；包括SEQ ID No.11(MAD2011核酸酶)、SEQ ID No.43(gRNA重复序列)和SEQID No.44(gRNA tracr序列)的MAD2011系统；包括SEQ ID No.11(MAD2011核酸酶)、SEQ IDNo.45(gRNA重复序列)和SEQ ID No.46(gRNA tracr序列)的MAD2011系统；和包括SEQ IDNo.11(MAD2011核酸酶)、SEQ ID No.47(gRNA重复序列)和SEQ ID No.48(gRNA tracr序列)的MAD2011系统。在一些方面，MAD系列系统组分作为待转录的序列(在gRNA组分的情况下)和待转录和翻译的序列(在mAD系列核酸酶的情况下)被递送，并且在一些方面，MAD系列核酸酶的编码序列和gRNA组分序列在同一载体上。在其他方面，MAD系列核酸酶的编码序列和gRNA组分序列在不同的载体上，并且在一些方面，gRNA组分序列位于编辑盒中，该编辑盒还包含供体DNA(例如，同源臂)。在其他方面，MAD系列核酸酶作为肽被递送到细胞或MAD系列核酸酶和gRNA组分作为核糖核酸酶复合体被递送到细胞。
另外，提供了包括以下的切口酶：MAD2001切口酶1[SEQ ID No.14]；MAD2001切口酶2[SEQ ID No.15]；失活的MAD2001[SEQ ID No.16]；MAD2007切口酶1[SEQ ID No.17]；MAD2007切口酶2[SEQ ID No.18]；失活的MAD2007[SEQ ID No.19]；MAD2008切口酶1[SEQID No.51]；MAD2008切口酶2[SEQ ID No.52]；失活的MAD2008[SEQ ID No.53]；MAD2009切口酶1[SEQ ID No.54]；MAD2009切口酶2[SEQ ID No.55]；失活的MAD2009[SEQ ID No.56]；MAD 2011切口酶1[SEQ ID No.57]；MAD2011切口酶2[SEQ ID No.58]；和失活的MAD2011[SEQ ID No.59]。
另外，提供了用于MAD2007的间隔区序列和PAM序列对，包括SEQ ID No.60和SEQID No.61；SEQ ID No.62和SEQ ID No.63；SEQ ID No.64和SEQ ID No.65；SEQ ID No.66和SEQ ID No.67；SEQ ID No.68和SEQ ID No.69；SEQ ID No.70和SEQ ID No.71；SEQ IDNo.72和SEQ ID No.73；SEQ ID No.74和SEQ ID No.75；SEQ ID No.76和SEQ ID No.77；SEQID No.78和SEQ ID No.79；SEQ ID No.80和SEQ ID No.81；SEQ ID No.82和SEQ ID No.83；SEQ ID No.84和SEQ ID No.85；SEQ ID No.86和SEQ ID No.87；SEQ ID No.88和SEQ IDNo.89；SEQ ID No.90和SEQ ID No.91；SEQ ID No.92和SEQ ID No.93；SEQ ID No.94和SEQID No.95；SEQ ID No.96和SEQ ID No.97；SEQ ID No.98和SEQ ID No.99；SEQ ID No.100和SEQ ID No.101；SEQ ID No.102和SEQ ID No.103；SEQ ID No.104和SEQ ID No.105；SEQID No.106和SEQ ID No.107；SEQ ID No.108和SEQ ID No.109；SEQ ID No.110和SEQ IDNo.111；SEQ ID No.112和SEQ ID No.113；SEQ ID No.114和SEQ ID No.115；SEQ IDNo.116和SEQ ID No.117；SEQ ID No.118和SEQ ID No.119；SEQ ID No.120和SEQ IDNo.121；SEQ ID No.122和SEQ ID No.123；SEQ ID No.124和SEQ ID No.125；以及SEQ IDNo.126和SEQ ID No.127。此外，提供了用于MAD2001的间隔区序列和PAM序列对，包括SEQID No.128和SEQ ID No.129；SEQ ID No.130和SEQ ID No.131；SEQ ID No.132和SEQ IDNo.133；SEQ ID No.134和SEQ ID No.135；SEQ ID No.136和SEQ ID No.137；以及SEQ IDNo.138和SEQ ID No.139。
在又另一实施方案中，提供了来自阿扎夏普氏菌(Sharpea azabuensis)的另外的MAD2007序列，包括SEQ ID No.142；SEQ ID No.143；和SEQ ID No.144。
下面更详细地描述本发明的这些方面和其他特征和优势。
附图简述
图1是用于创建和筛选挖掘的MAD系列核酸酶或RGN的示例性工作流程。
图2是发现的新颖的核酸酶的蛋白质大小对比搜索评分的图。
图3是新的MAD核酸酶的gRNA设计的示意图。
图4以序列标志(sequence logo)的形式显示了被挖掘的MAD系列核酸酶裂解的靶的富集。
图5是显示MAD2001、MAD2007、MAD2008、MAD2009和MAD2011在HEK293T细胞中的活性的一系列柱状图。
图6显示了MAD2001、MAD2007、MAD2008、MAD2009和MAD2011的人类基因组覆盖。
图7显示了GFP-HEK293T细胞相对于阴性对照的百分比。
图8是HEK293T细胞中MAD2007的PAM的序列标志。
图9显示了GFP-HEK293T细胞相对于阴性对照的百分比。
图10显示了两种不同的人类密码子优化的MAD2007核酸酶在HEK293T细胞中功能丧失的百分比。
详细描述
下文结合附图阐述的描述意图描述所公开主题的多种说明性实施方案。结合每种说明性实施方案描述了特定特征和功能；然而，对本领域技术人员将明显的是，所公开的实施方案可以在没有这些特定特征和功能的每一种的情况下实践。此外，除非明确声明或者特征或功能与另外的实施方案不兼容，否则结合一种实施方案描述的所有功能意图适用于本文描述的另外的实施方案。例如，除非特征或功能与替代实施方案不兼容，否则在结合一种实施方案明确描述给定特征或功能但没有结合替代实施方案明确提及的情况下，应当理解，该特征或功能可以结合替代实施方案来部署、利用或实现。
除非另外指出，否则本文描述的技术的实践可以采用有机化学、聚合物技术、分子生物学(包括重组技术)、细胞生物学、生物化学、生物乳液产生和测序技术的常规技术和描述，这些都在本领域从业的人员的技术内。这样的常规的技术包括聚合物阵列合成、多核苷酸的杂交和连接以及使用标记物的杂交检测。合适的技术的具体的说明可以通过参考本文的实例获得。然而，当然，也可以使用其他等同的常规程序。这样的常规技术和描述可以见于标准实验室手册，诸如Green等人编著(1999),Genome Analysis:A Laboratory ManualSeries(卷I-IV)；Weiner,Gabriel,Stephens,编著(2007),Genetic Variation:ALaboratory Manual；Dieffenbach,Dveksler,编著(2003),PCR Primer:A LaboratoryManual；Bowtell和Sambrook(2003),DNA Microarrays:A Molecular Cloning Manual；Mount(2004),Bioinformatics:Sequence and Genome Analysis；Sambrook和Russell(2006),Condensed Protocols from Molecular Cloning:A Laboratory Manual；以及Sambrook和Russell(2002),Molecular Cloning:A Laboratory Manual(全部来自ColdSpring Harbor Laboratory Press)；Stryer,L.(1995)Biochemistry(第4版)W.H.Freeman,New York N.Y.；Gait,“Oligonucleotide Synthesis:A PracticalApproach”1984,IRL Press,London；Nelson和Cox(2000),Lehninger,Principles ofBiochemistry第3版,W.H.Freeman Pub.,New York,N.Y.；Berg等人(2002)Biochemistry,第5版,W.H.Freeman Pub.,New York,N.Y.；Cell and Tissue Culture:LaboratoryProcedures in Biotechnology(Doyle&Griffiths,编著,John Wiley&Sons 1998)；Mammalian Chromosome Engineering–Methods and Protocols(G.Hadlaczky,编著,Humana Press2011)；Essential Stem Cell Methods,(Lanza和Klimanskaya,编著,Academic Press 2011)，所有文献出于所有目的通过引用以其整体并入本文。核酸酶特异性技术可以见于，例如，Genome Editing and Engineering From TALENs and CRISPRs toMolecular Surgery,Appasani和Church,2018；以及CRISPR:Methods and Protocols,Lindgren和Charpentier,2015；这两篇文献出于所有目的通过引用以其整体并入本文。酶工程化的基本方法可以见于，Enzyme Engineering Methods and Protocols,Samuelson,编著,2013；Protein Engineering,Kaumaya,编著,(2012)；以及Kaur和Sharma,“DirectedEvolution:An Approach to Engineer Enzymes”,Crit.Rev.Biotechnology,26:165-69(2006)。
注意，除非上下文另外清楚指示，如本文和所附的权利要求书中使用的，单数形式“一(a)”、“一(an)”和“该(the)”包括复数指代物。因此，例如，提及“寡核苷酸”是指一种或更多种寡核苷酸。术语诸如“第一”、“第二”、“第三”等，仅标识如本文公开的许多部分、组件、步骤、操作、功能和/或参考点中的一个，并且同样不必然将本公开内容的实施方案限制为任何特定的配置或方向。
除非另外定义，否则本文使用的所有技术和科学术语具有与本发明所属领域内的普通技术人员通常理解的相同含义。本文提及的所有的出版物为了描述和公开可以与本文描述的发明结合使用的设备、方法和细胞群体的目的，以引用方式并入。
在提供值的范围情况下，应理解，在该范围的上限值和下限值之间的每一个中间值和该规定的范围内的任何其他规定的值或中间值被涵盖在本发明内。这些较小的范围的上限值和下限值可以独立地被包括在较小的范围内，并且也被涵盖在本发明内，受限于规定的范围内的任何特定地排除的限值。在规定的范围包括限值中的一个或两个的情况下，将那些所包括的限值中的任一个或两个排除的范围也被包括在本发明中。
在以下的描述中，阐述了许多具体细节，以提供对本发明的更充分理解。然而，对本领域普通技术人员将明显的是，可以在没有一个或更多个这些具体细节的情况下，实践本发明。在其他情况下，为了避免使本发明含混不清，尚未描述本领域技术人员熟知的特征和熟知的程序。
如本文使用的术语“互补”是指核苷酸之间的Watson-Crick碱基配对，并且特别地是指彼此形成氢键的核苷酸，其中胸腺嘧啶或尿嘧啶残基通过两个氢键与腺嘌呤残基连接，并且胞嘧啶和鸟嘌呤残基通过三个氢键连接。通常，核酸包含被描述为与指定的第二核苷酸序列具有“互补性百分比”或“同源性百分比”的核苷酸序列。例如，核苷酸序列可以与指定的第二核苷酸序列具有80％、90％或100％的互补性，指示序列的10个核苷酸中的8个、10个核苷酸中的9个或10个核苷酸中的10个与指定的第二核苷酸序列互补。例如，核苷酸序列3’-TCGA-5’与核苷酸序列5’-AGCT-3’是100％互补的；并且核苷酸序列3’-TCGA-5’与核苷酸序列5’-TAGCTG-3’的区域是100％互补的。
术语DNA“控制序列”统指启动子序列、多腺苷酸化信号、转录终止序列、上游调控结构域、复制起点、内部核糖体进入位点、核定位序列、增强子等，它们共同地提供编码序列在接受者细胞中的复制、转录和翻译。只要选择的编码序列能够在适当的宿主细胞中被复制、转录和(对于一些组分)翻译，则并非所有这些类型的控制序列都需要存在。
如本文使用的，术语“供体DNA”或“供体核酸”是指被设计成通过使用核酸引导的核酸酶的同源重组将DNA序列修饰(插入、缺失、取代)引入基因座的核酸。对于同源指导的修复，供体DNA必须与基因组靶序列中的“切割位点”或待编辑位点侧翼的区具有足够的同源性。一条或更多条同源臂的长度将取决于，例如，所做修饰的类型和大小。在许多情况下，并且优选地，供体DNA将与基因组靶基因座具有两个序列同源性区(例如，两个同源臂)。优选地，“插入物(insert)”区或“DNA序列修饰”区(期望引入细胞中的基因组靶基因座的核酸修饰)将位于两个同源区之间。DNA序列修饰可以改变一个特定位点或多于一个特定位点处的靶基因组DNA序列的一个或更多个碱基。改变可以包括改变靶序列的1个、2个、3个、4个、5个、10个、15个、20个、25个、30个、35个、40个、50个、75个、100个、150个、200个、300个、400个或500个或更多个碱基对。缺失或插入可以是靶序列的1个、2个、3个、4个、5个、10个、15个、20个、25个、30个、40个、50个、75个、100个、150个、200个、300个、400个或500个或更多个碱基对的缺失或插入。
术语“引导核酸”或“引导RNA”或“gRNA”是指包含以下的多核苷酸：1)能够与基因组靶基因座杂交的引导序列和2)能够与核酸引导的核酸酶相互作用或复合的支架序列(参见例如，图1)。
“同源性”或“同一性”或“相似性”是指两个肽之间的序列相似性，或者在本公开内容的上下文中，更常见地是指两个核酸分子之间的序列相似性。术语“同源区”或“同源臂”是指供体DNA上与靶基因组DNA序列具有一定程度同源性的区域。同源性可以通过比较每个序列中的位置来确定，所述每个序列可以出于比较的目的而被对齐。当比较的序列中的一个位置被相同的碱基或氨基酸占据时，那么分子在该位置处是同源的。序列之间的同源性程度随着序列共有的匹配或同源位置数而变化。
“可操作地连接”指其中如此描述的组分被配置以执行它们的通常功能的元件布置。因此，可操作地连接至编码序列的控制序列能够影响编码序列的转录，并且在一些情况下，能够影响编码序列的翻译。只要控制序列起作用以指导编码序列的表达，控制序列不必与编码序列邻接。因此，例如，不翻译但转录的间插序列可以存在于启动子序列和编码序列之间，且启动子序列仍可以被认为是“可操作地连接”至编码序列。事实上，这样的序列不必驻留于同一连续DNA分子(即染色体)上，并且仍可以具有引起调控改变的相互作用。
“启动子”或“启动子序列”是能够与RNA聚合酶结合并启动多核苷酸或多肽编码序列(诸如信使RNA、核糖体RNA、小核RNA(small nuclear RNA)或核仁小RNA(smallnucleolar RNA)、引导RNA或由任何类别的任何RNA聚合酶I、II或III转录的任何种类的RNA)的转录的DNA调控区。启动子可以是组成型或诱导型，并且在一些实施方案中，特别地在许多采用选择的实施方案中，核酸引导的核酸酶编辑系统的至少一种组分的转录处于诱导型启动子的控制下。
如本文使用的术语“选择标记(selectable marker)”是指引入细胞中的、赋予适于人工选择的性状的基因。一般使用的选择标记是本领域普通技术人员熟知的。可以采用药物选择标记，诸如氨苄青霉素/羧苄青霉素、卡那霉素、氯霉素、红霉素、四环素、庆大霉素、博莱霉素、链霉素、鼠李糖、嘌呤霉素、潮霉素、杀稻瘟素和G418。在其他实施方案中，选择标记包括但不限于人类神经生长因子受体(用MAb检测，诸如美国专利第6,365,373号中描述的)；截短的人类生长因子受体(用MAb检测)；突变体人类二氢叶酸还原酶(DHFR；可用荧光MTX底物)；分泌型碱性磷酸酶(SEAP；可用荧光底物)；人类胸苷酸合酶(TS；赋予对抗癌剂氟脱氧尿苷的抗性)；人类谷胱甘肽S-转移酶α(GSTA1；将谷胱甘肽与干细胞选择性烷化剂白消安缀合；CD34+细胞中的化学保护性选择标记)；造血干细胞中的CD24细胞表面抗原；赋予对N-膦酰乙酰基-L-天冬氨酸(PALA)的抗性的人类CAD基因；人类多耐药性-1(MDR-1；可通过增加的耐药性选择或通过FACS富集P-糖蛋白表面蛋白)；人类CD25(IL-2α；可通过Mab-FITC检测)；甲基鸟嘌呤-DNA甲基转移酶(MGMT；可通过卡莫司汀(carmustine)选择)；和胞苷脱氨酶(CD；可通过Ara-C选择)。如本文使用的“选择性培养基”是指向其中添加了选择选择标记或针对选择标记进行选择的化学化合物或生物部分的细胞生长培养基。
术语“靶基因组DNA序列”、“靶序列”或“基因组靶基因座”是指体外或体内，或者细胞或细胞群体的核酸(例如基因组)中的期望使用核酸引导的核酸酶编辑系统对至少一个核苷酸进行改变的任何基因座。靶序列可以是基因组基因座或染色体外基因座。
“载体”是包含待递送至细胞和/或在细胞中表达的期望的一种序列或更多种序列的多种核酸中的任一种。载体通常由DNA构成，但是RNA载体也是可用的。载体包括但不限于质粒、F粘粒(fosmid)、噬菌粒、病毒基因组、合成染色体等。如本文使用的，短语“引擎载体”包含用于本公开内容的核酸引导的核酸酶系统和方法中的核酸酶的编码序列。在细菌系统中，引擎载体还可以包含λRed重组工程系统或其等同物。引擎载体通常还包含选择标记。如本文使用的，短语“编辑载体”包含供体核酸和gRNA编码序列，所述供体核酸任选地包括对靶序列的改变，所述改变在编辑发生后阻止核酸酶在靶序列中的PAM或间隔区(spacer)处结合。编辑载体还可以包括选择标记和/或条形码。在一些实施方案中，可以将引擎载体和编辑载体组合；即，引擎载体的内容物可以在编辑载体上找到。此外，引擎载体和编辑载体包含可操作地连接至例如核酸酶编码序列、重组工程系统编码序列(如果存在)、供体核酸、引导核酸和一种或更多种选择标记的控制序列。
核酸引导的核酸酶在基因组系统中的编辑
RNA引导的核酸酶(RGN)已迅速成为用于原核生物和真核生物基因组工程化的基础工具。成簇的快速间隔短回文重复序列(CRISPR)系统是保护原核生物抵抗可移动遗传元件(MGE)的适应性免疫系统。RGN是这一防御系统的主要部分，因为它们鉴定并摧毁MGE。RGN可以通过使将RGN引导到特定的靶DNA的CRISPR RNA(crRNA)重编程来重新用于各种生物体的基因组编辑。到目前为止，已经为各种应用鉴定了许多不同的RGN；然而，有对于特定的应用使得一些RGN比其他RGN更受期望的各种特性。RGN可以用于创建特定的双链断裂(DSB)、一条DNA链的特定切口或引导另一个部分到特定的DNA序列。
RGN特异性靶向任何基因组序列的能力可能是RGN最受期望的特征；然而，只有在靶DNA还含有称为PAM(前间区邻近基序)的短基序时，RGN才能接近它们期望的靶，PAM对于每个RGN是特异的。V型RGN诸如MAD7、AsCas12a和LbCas12a倾向于接近在5’末端含有YTTN/TTTN的DNA靶，而II型RGN靶向在3’末端含有特定短基序的DNA序列。本领域中II型RGN的一个熟知的实例是SpCas9，它需要靶DNA的3’末端的NGG。II型RGN具有相对于V型RGN明显不同的结构域结构。此外，II型RGN除了crRNA外，还需要反式激活RNA(tracrRNA)以获得最佳功能。与V型RGN相比，II型RGN创建更接近PAM序列的双链断裂，这对于精确的基因组编辑应用是高度期望的。
迄今已发现许多II型RGN；然而，它们在广泛应用中的使用受到限制性PAM的限制。例如，SpCas9的PAM在基因组中AT丰富的区域出现的频率较低。具有新的和较少限制性的PAM的新的RGN对该领域是有益的。此外，并不是所有的II型核酸酶在多种生物体中都是有活性的。例如，科学文献中已经讨论了许多RGN，但只有少数被证明在体外是有活性的，并且在细胞中，特别是在哺乳动物细胞中有活性的RGN更少。本公开内容鉴定了具有新颖的PAM并且在哺乳动物细胞中是有活性的多于一种RGN。
在进行核酸引导的核酸酶编辑时，可以将挖掘的MAD系列核酸酶或RGN作为多肽递送到待编辑的细胞；可选地，将编码挖掘的MAD系列核酸酶的多核苷酸序列转化或转染到待编辑的细胞中。编码挖掘的MAD系列核酸酶的多核苷酸序列可以针对特定细胞诸如古核生物、原核细胞或真核细胞中的表达被密码子优化。真核细胞可以是酵母、真菌、藻类、植物、动物或人类的细胞。真核细胞可以是特定生物体的细胞或来源于特定生物体的细胞，所述特定生物体诸如哺乳动物，包括但不限于人类、小鼠、大鼠、兔、犬或非人类哺乳动物，包括非人类灵长类动物。待采用的挖掘的MAD系列核酸酶的选择取决于许多因素，诸如在靶序列中待进行何种类型的编辑，以及适当的PAM是否位于期望的靶序列附近。挖掘的MAD系列核酸酶可以由载体(例如，引擎载体)上的DNA序列编码，并且处于组成型或诱导型启动子的控制下。在一些实施方案中，编码核酸酶的序列处于诱导型启动子的控制下，并且该诱导型启动子可以与控制引导核酸转录的诱导型启动子分开但相同；即，分开的诱导型启动子可驱动核酸酶和引导核酸序列的转录，但是这两个诱导型启动子可以是相同类型的诱导型启动子(例如，二者都是pL启动子)。可选地，控制核酸酶表达的诱导型启动子可以与控制引导核酸转录的诱导型启动子不同；即，例如，核酸酶可以处于pBAD诱导型启动子的控制下，并且引导核酸可以处于pL诱导型启动子的控制下。
通常，引导核酸(例如，gRNA)可以与相容的核酸引导的核酸酶复合，并且然后可以与靶序列杂交，从而将核酸酶指导至靶序列。对于本文描述的MAD系列酶，核酸引导的核酸酶编辑系统使用组合并起引导核酸的作用的两个单独的引导核酸组分；即CRISPR RNA(crRNA)和反式激活CRISPR RNA(tracrRNA)。gRNA可以由多核苷酸分子诸如质粒、线性构建体上的DNA序列编码，或者编码序列可以驻留于编辑盒内，并且处于组成型启动子的控制下，或者在一些实施方案中，在如下文描述的诱导型启动子的控制下。
引导核酸包含与靶序列具有足够互补性以与靶序列杂交并且指导复合的核酸引导的核酸酶与靶序列的序列特异性结合的引导多核苷酸序列。引导序列和对应的靶序列之间的互补性程度在使用合适的比对算法进行最佳比对时是约或多于约50％、60％、75％、80％、85％、90％、95％、97.5％、99％或更多。最佳比对可以通过使用用于比对序列的任何合适的算法来确定。在一些实施方案中，引导序列的长度是约或多于约10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、20个、21个、22个、23个、24个、25个、26个、27个、28个、29个、30个、35个、40个、45个、50个、75个或更多个核苷酸。在一些实施方案中，引导序列的长度为少于约75个、50个、45个、40个、35个、30个、25个、20个核苷酸。优选地，引导序列是10-30个或15-20个核苷酸长，或者长度是15个、16个、17个、18个、19个或20个核苷酸。
在本发明的方法和组合物中，引导核酸的组分作为待由质粒或载体表达的序列提供，并且包含引导序列和支架序列二者作为启动子控制下的，并且在一些实施方案中，诱导型启动子控制下的单一转录物。通常，为了在靶序列中产生编辑，gRNA/核酸酶复合体与如由引导RNA确定的靶序列结合，并且核酸酶识别与靶序列邻近的前间区邻近基序(PAM)序列。靶序列可以是对原核细胞或真核细胞而言为内源或外源的任何多核苷酸，或体外的任何多核苷酸。例如，靶序列可以是驻留于真核细胞的细胞核中的多核苷酸。靶序列可以是编码基因产物(例如，蛋白)的序列或非编码序列(例如，调控多核苷酸、内含子、PAM或“垃圾”DNA(“junk”DNA))。
引导核酸可以是编码供体核酸的编辑盒的一部分。可选地，引导核酸可以不是编辑盒的一部分，而是可以被编码在引擎载体或编辑载体骨架上。例如，可以首先将编码引导核酸的序列组装或插入载体骨架中，随后将供体核酸插入例如编辑盒中。在其他情况下，可以首先将例如编辑盒中的供体核酸插入或组装到载体骨架中，随后插入编码引导核酸的序列。在又其他情况下，编码引导核酸和供体核酸(例如，插入编辑盒中)的序列同时但分开插入或组装到载体中。在又其他实施方案中，编码引导核酸的序列和编码供体核酸的序列二者均包含在编辑盒中。
靶序列与PAM相关，PAM是由gRNA/核酸酶复合体识别的短核苷酸序列。用于不同的核酸引导的核酸酶的精确的PAM序列和长度要求不同；然而，PAM通常是与靶序列邻近或接近的2-7个碱基对序列，并且取决于核酸酶，可以在靶序列的5’或3’。核酸引导的核酸酶的PAM相互作用结构域的工程化可以允许改变PAM特异性、改进保真度或降低保真度。在某些实施方案中，靶序列的基因组编辑既将期望的DNA改变引入靶序列，例如细胞的基因组DNA，又去除靶序列中的前间区突变(PAM)区，使靶序列中的前间区突变(PAM)区突变或失活。使靶序列处的PAM失活排除了对该靶序列处细胞基因组的另外的编辑，例如，在后续的编辑轮中随后暴露于与合成的引导核酸复合的核酸引导的核酸酶时。因此，具有期望的靶序列编辑和改变的PAM的细胞可以使用与合成的引导核酸复合的核酸引导的核酸酶来选择，所述合成的引导核酸与靶序列互补。没有经历第一编辑事件的细胞会被切割，引起双链DNA断裂，并且因此会无法继续存活。包含期望的靶序列编辑和PAM改变的细胞不会被切割，因为这些编辑的细胞不再包含必需的PAM位点，并且会继续生长和繁殖。
如以上提及的，核酸引导的核酸酶可以识别的靶序列的范围受将特定PAM定位于期望的靶序列附近的需要限制。因此，以基因组编辑必需的精度靶向编辑通常可能是困难的。已经发现，核酸酶可以非常好地识别一些PAM(例如，典型PAM(canonical PAM))，而不太好或较差地识别其他PAM(例如，非典型PAM)。因为本文公开的挖掘的MAD系列核酸酶可以识别不同的PAM，所以挖掘的MAD系列核酸酶增加了可以被靶向用于编辑的靶序列的数量；即，挖掘的MAD系列核酸酶减少了基因组中“PAM沙漠(PAM deserts)”区域。因此，挖掘的MAD系列核酸酶通过增加被识别的PAM序列的数量(多样化(variety))扩大了可以被编辑的靶序列的范围。此外，可以将挖掘的MAD系列核酸酶的混合物(cocktail)递送至细胞，使得可以在单次编辑运行中编辑与若干不同PAM邻近的靶序列。
核酸引导的核酸酶系统的另一组分是供体核酸。在一些实施方案中，供体核酸与引导核酸在同一多核苷酸(例如，编辑载体或编辑盒)上，并且可以(但不必然)处于与引导核酸相同的启动子的控制下(例如，驱动引导核酸和供体核酸二者转录的单一启动子)。关于这种类型的盒，参见USPN10,240,167；USPN 10,266,849；USPN 9,982,278；USPN 10,351,877；USPN10,364,442；USPN 10,435,715；和USPN 10,465,207。供体核酸被设计成用作用于与靶序列同源重组的模板，该靶序列被作为gRNA/核酸酶复合体的一部分的核酸引导的核酸酶切口或裂解。供体核酸多核苷酸可以具有任何合适的长度，诸如约或多于约20个、25个、50个、75个、100个、150个、200个、500个或1000个核苷酸的长度。在某些优选的方面，供体核酸可以以20-300个核苷酸之间，更优选地50-250个核苷酸之间的寡核苷酸提供。供体核酸包含与靶序列的一部分互补的区域(例如同源臂)。当最佳比对时，供体核酸与靶序列重叠(互补)例如约20个、25个、30个、35个、40个、50个、60个、70个、80个、90个或更多个核苷酸。在许多实施方案中，供体核酸包含位于供体核酸和靶模板之间的突变或差异的侧翼的两个同源臂(与靶序列互补的区域)。供体核酸包含与靶序列相比的至少一个突变或改变，诸如与靶序列相比的插入、缺失、修饰或其任何组合。
通常，供体核酸以编辑盒提供，其被插入到载体骨架中，其中载体骨架可以包含驱动gRNA转录的启动子和gRNA编码序列，或者载体骨架可以包含驱动gRNA转录的启动子，但不包含gRNA本身。此外，可以有多于一个，例如两个、三个、四个或更多个引导核酸/供体核酸盒插入到引擎载体中，其中每个引导核酸处于分开的不同启动子、分开的相似启动子的控制下，或者其中所有引导核酸/供体核酸对处于单个启动子的控制下。在一些实施方案中，驱动gRNA和供体核酸(或驱动多于一个gRNA/供体核酸对)转录的启动子是诱导型启动子。诱导型编辑的优点在于，分离的细胞可以生长几倍至许多倍的细胞倍增以在启动编辑前建立集落，这增加具有编辑的细胞将存活的可能性，因为由有效编辑(active editing)引起的双链切割对细胞有很大毒性。这种毒性既导致编辑的集落中的细胞死亡，也导致确实存活但必须在编辑后修复和恢复(recover)的编辑的细胞生长延滞。然而，在编辑的细胞具有恢复的机会后，编辑的细胞集落的尺寸最终会赶上未编辑的细胞集落的尺寸。例如参见USPN 10,533,152；USPN 10,550,363；USPN 10,532,324；和2019年10月9日提交的USSN16/597,826；2019年10月9日提交的USSN 16/597,831；2019年11月25日提交的USSN 16/693,630；2019年11月18日提交的USSN 16/687,640；和2019年11月15日提交的USSN 16/686,066。此外，引导核酸可以有效指导编辑盒中多于一个供体核酸的编辑；例如，如果期望的编辑在靶序列中彼此接近。
除了供体核酸之外，编辑盒可以包含一个或更多个引物位点。引物位点可以用于通过使用寡核苷酸引物扩增编辑盒；例如，如果引物位点位于编辑盒的一个或更多个其他组分的侧翼。
此外，编辑盒可以包含条形码。条形码是对应于供体DNA序列的独特DNA序列，使得条形码可以鉴定对对应靶序列进行的编辑。条形码通常包含四个或更多个核苷酸。在一些实施方案中，编辑盒包含代表例如供体核酸的全基因或全基因组文库的供体核酸的集合。编辑盒的文库被克隆到载体骨架中，其中，例如，每个不同的供体核酸与不同的条形码相关。
此外，在一些实施方案中，编码核酸引导的核酸酶系统的组分的表达载体或盒还编码一个或更多个细胞核定位序列(NLS)，诸如约或多于约1个、2个、3个、4个、5个、6个、7个、8个、9个、10个或更多个NLS。在一些实施方案中，核酸酶包含在挖掘的MAD系列RGN的氨基末端处或附近的NLS，在挖掘的MAD系列RGN的羧基末端处或附近的NLS，或组合。
引擎载体和编辑载体包含可操作地连接至待转录的组分序列的控制序列。如以上陈述的，驱动挖掘的MAD系列核酸酶编辑系统的一种或更多种组分转录的启动子可以是诱导型的，并且如果要进行选择，可能采用诱导型系统。已经开发了用于在植物、微生物和动物细胞(包括哺乳动物细胞)中控制基因表达的许多基因调控控制系统，包括pL启动子(通过CI857阻遏物的热失活诱导)、pBAD启动子(通过将阿拉伯糖添加至细胞生长培养基中诱导)和鼠李糖诱导型启动子(通过将鼠李糖添加至细胞生长培养基中诱导)。其他系统包括四环素控制的转录激活系统(Tet-On/Tet-Off，Clontech,Inc.(Palo Alto,CA)；Bujard和Gossen,PNAS,89(12):5547-5551(1992))、Lac开关诱导型系统(Wyborski等人，EnvironMol Mutagen,28(4):447-58(1996)；DuCoeur等人，Strategies 5(3):70-72(1992)；美国专利第4,833,080号)、蜕皮素诱导型基因表达系统(No等人，PNAS,93(8):3346-3351(1996))、cumate基因开关系统(Mullick等人，BMC Biotechnology,6:43(2006))、以及他莫昔芬诱导型基因表达(Zhang等人，Nucleic Acids Research,24:543-548(1996))以及其他。
通常，在活细胞中进行基因组编辑要求用进行核酸引导的核酸酶编辑所必需的组分转化细胞。例如，细胞可以用分开的引擎载体和编辑载体同时转化；细胞可能已经表达挖掘的MAD系列核酸酶(例如，细胞可能已经用引擎载体转化，或者挖掘的MAD系列核酸酶的编码序列可以稳定地整合到细胞基因组中)，使得仅需要将编辑载体转化到细胞中；或者细胞可以用包含进行核酸引导的核酸酶基因组编辑所需的所有组分的单个载体转化。
可以使用各种递送系统将核酸引导的核酸酶编辑系统组分引入(例如，转化或转染)宿主细胞。这些递送系统包括酵母系统、脂质体转染系统、显微注射系统、基因枪系统、病毒微体(virosome)、脂质体、免疫脂质体、聚阳离子、脂质:核酸缀合物、病毒粒子(virion)、人工病毒粒子、病毒载体、电穿孔、细胞可渗透肽、纳米粒子、纳米线、外泌体(exosome)的使用。可选地，可以使用分子特洛伊木马(trojan horse)脂质体跨越血脑屏障递送核酸引导的核酸酶组分。特别感兴趣的是电穿孔的使用，特别是流通式电穿孔(作为独立的仪器或作为自动化多模块系统中的模块)的使用，如以下中描述的：USPN 10,435,713；10,443,074；10,323,258；和10,415,058。
在将细胞用进行核酸引导的核酸酶编辑所必需的组分转化后，使细胞在促进编辑的条件下培养。例如，使用如果组成型启动子驱动挖掘的MAD系列核酸酶和/或gRNA的转录，转化的细胞仅需要在典型条件下(例如，温度、CO2气氛等)在典型培养基中培养。可选地，如果编辑是诱导型的——例如通过激活诱导型启动子，该诱导型启动子控制核酸引导的核酸酶编辑所需的一种或更多种组分的转录，诸如，例如gRNA、供体DNA、核酸酶的转录，或者，在细菌的情况下，编辑是重组工程系统诱导型的——则细胞经历诱导条件。
实施例
提出以下实施例，以便为本领域技术人员提供如何制备和使用本发明的完整公开和描述，且以下实施例并不意图限制发明人视作其发明的范围，它们也不意图代表或暗示下文的实验是进行的所有实验或仅有的实验。本领域的技术人员应该明白，可以在对具体方面中所示的发明作出许多变化和/或修饰而不脱离本发明所广泛描述的精神或范畴。因此，本文的方面在各个方面中被认为是说明性的而非限制性的。
实施例1：示例性工作流程概述
图1示出了用于创建和用于体外筛选挖掘的MAD系列酶的示例性工作流程100。在第一步骤101，制备并克隆载体以制备向其中插入挖掘的MAD系列RGN的编码序列的模板载体。在另一步骤103，执行宏基因组挖掘以基于例如序列、潜在PAM和活性的可能性来鉴定推定的感兴趣RGN。在计算机模拟鉴定推定的感兴趣RGN后，构建盒105并克隆到载体骨架中，并且然后转化到细胞中，从而产生挖掘的MAD系列RGN的文库。用挖掘的MAD系列RGN转化的细胞排列在96孔板中107，用于储存。
在步骤109，取来自每个孔的细胞的等分试样，并从每个等分试样扩增挖掘的MAD系列RGN。平行地，对于每个挖掘的MAD系列RGN，扩增gRNA文库110。在步骤111，将表达gRNA文库的扩增的PCR片段与扩增的挖掘的MAD序列RGN组合，以进行体外转录和翻译来制备活性核糖核酸酶蛋白复合体113。构建合成的靶文库115，其中测试每一个挖掘的MAD系列RGN的靶耗尽117。在靶耗尽之后，使用下一代测序产生扩增子用于分析119，并执行测序数据分析121以确定靶耗尽。
实施例2：宏基因组挖掘
使用来自不同来源的宏基因组组装基因组(MAG)，包括Genbank Bioproject登录号PRJNA348753、PRJNA385857、PRJNA432584和PRJNA434545以使用HMMER隐马尔可夫模型搜索寻找新的、推定的CRISPR核酸酶。鉴定了数百种潜在的核酸酶。图2显示了发现的新颖的RGN，将蛋白质大小对比HMMER搜索评分绘图。对于每一个存在核酸酶的MAG，鉴定推定的CRISPR阵列并提取间隔区序列。然后使用这些间隔区作为查询来搜索JGI IMG/VR病毒宏基因组数据库(Paez-Espino等人,Nucleic Acids Res.2017Jan 4；45(D1):D457-D465)并根据邻近间隔区命中的病毒序列预测推定的PAM序列。根据序列、潜在PAM和核酸酶可能有活性的置信度，鉴定出13种核酸酶(表1)用于体外验证。13种核酸酶中每一种的序列示于表2。
表1
Figure BDA0003688304840000191
表2
Figure BDA0003688304840000192
Figure BDA0003688304840000201
Figure BDA0003688304840000211
Figure BDA0003688304840000221
Figure BDA0003688304840000231
Figure BDA0003688304840000241
Figure BDA0003688304840000251
Figure BDA0003688304840000261
Figure BDA0003688304840000271
实施例3：载体克隆、MAD系列RGN文库构建及PCR
将挖掘的MAD系列RGN编码序列克隆到pUC57载体中，其中T7启动子序列附接到编码序列的5’末端，并且T7终止子序列附接到编码序列的3’末端。将100ng质粒混合物转化到E.
Figure BDA0003688304840000272
SUPREME电感受态solo细胞(Lucigen,Middleton,WI)中。将细胞在振荡培养箱中在37℃在5mL恢复培养基中恢复1小时后，添加1mL 50％甘油，并且将细胞以100μL等分试样储存在-80℃。
将储存的细胞在磷酸盐缓冲盐水中稀释，并且铺展在具有100μg/mL羧苄青霉素的LB琼脂板上。然后将细胞在培养箱中在37℃生长过夜。挑取菌落并接种到96孔培养块中的1mL LB培养基(100μg/mL羧苄青霉素)中。将培养物在振荡培养箱中在37℃生长过夜。接下来，将1μL细胞稀释到500μL PCR级水中，并且将稀释培养物的25μl等分试样使用热循环仪在95℃煮沸5min。使用细胞来PCR扩增不同的挖掘的MAD系列RGN编码序列。将剩余的培养物添加浓度为10％v/v的甘油、储存在-80℃。
首先，使用煮沸的细胞作为挖掘的MAD系列RGN模板的来源，使用Q5热启动2x主混合物(master mix)试剂(NEB,Ipswich,MA)扩增挖掘的MAD系列RGN序列。正向引物5’-TTGGGTAACGCCAGGGTTTT[SEQ ID No.49]和反向引物5’-TGTGTGGAATTGTGAGCGGA[SEQ IDNo.50]扩增了pUC57载体中挖掘的MAD系列RGN侧翼的序列，包括分别在挖掘的MAD系列RGN的5’-末端和3’-末端的T7-启动子和T7-终止子组分。在96孔PCR板中的使用3.3μL煮沸的细胞样品作为模板的10μL PCR反应中，使用1μM引物。使用表3所示的PCR条件：
表3：
步骤 温度 时间
变性 98℃ 30SEC
30个循环 98℃ 10SEC
66℃ 30SEC
72℃ 2.5MIN
最终延伸 72℃ 2MIN
保持 12℃
实施例4：gRNA文库构建
与每个RGN相关的功能性gRNA可能很难预测，因为RGN可能需要多于一个RNA才能发挥作用，并且转录的RNA的长度也可能高度可变。因此，为每个RGN创建了384个成员的gRNA文库。gRNA由可变间隔区序列、CRISPR重复序列、接头序列和tracrRNA序列组成。图3是挖掘的MAD系列RGN的gRNA设计的示意图。通过鉴定反义重复(anti-repeat)序列，发现了tracrRNA。crRNA与tracrRNA用GAAA接头共价连接。通过截短5’区、3’区和重复/反义重复双链体来创建gRNA文库，优化了初始的gRNA设计。为了发现最佳gRNA长度，包括了不同长度的间隔区、重复:反义重复双链体和tracrRNA的3’末端。根据gRNA的总长度，文库还被细分为六个池。这使得能够鉴定核酸酶活性最佳的最短gRNA。然后将这些gRNA克隆到T7启动子下游。
靶文库的设计是基于这些核酸酶的PAM将位于3’末端的假设。选择了两种GC含量不同的人工前间区。由于PAM序列的长度可以是3-7个或3-10个核苷酸的范围，因此为每个前间区制备了三个不同的PAM靶文库。文库1包含PAM NNNNATGC；文库2由PAM ATNNNNGC组成；并且文库3包含PAM ATGCNNNN。滑动PAM文库确保捕获范围从4nt到8nt的PAM。将靶文库克隆到包含对未切割靶进行下一代测序必需的序列的靶质粒中。
实施例5：用于产生MAD系列核酸酶和gRNA的体外转录和翻译
通过体外转录和翻译(txtl)测试MAD系列RGN的活性。gRNA质粒和核酸酶质粒二者都被包含在每个txtl反应中。使用
Figure BDA0003688304840000291
体外蛋白合成试剂盒(NEB,Ipswich,MA)从PCR扩增的MAD系列RGN文库产生挖掘的MAD系列RGN，并且还产生gRNA文库。在96孔板的每个孔中，混合表4中列出的试剂以开始产生挖掘的MAD7系列RGN和gRNA：
表4：
试剂 体积(μl)
1 SolA(NEB试剂盒) 3.3
2 SolB(NEB试剂盒) 2.5
3 PCR扩增的gRNA子池 1
4 鼠RNA酶抑制剂(NEB) 0.2
5 水 0.3
6 PCR扩增的T7 MAD系列RGN 1.0
将具有除了PCR扩增的T7-MAD系列RGN之外的所有试剂的主混合物在冰上混合，以覆盖足够的96孔板用于测定。在96孔板的每个孔中分配7.3μL的主混合物后，添加1μL在T7启动子控制下的PCR扩增的MAD系列RGN。将96孔板密封，并且在热循环仪中在37℃孵育4小时。将板保持在室温，直至添加靶池以进行靶耗尽反应。
孵育4小时允许产生挖掘的MAD系列RGN和gRNA后，将4μL的靶文库池(10ng/μL)添加至体外转录/翻译反应混合物中并允许在37℃耗尽30分钟、3小时或过夜。将靶耗尽反应混合物稀释到包含RNA酶A的PCR级水中，并且然后在95℃煮沸5min。然后扩增混合物并测序。PCR条件示于表5：
表5
步骤 温度 时间
变性 98℃ 30SEC
6个循环 98℃ 10SEC
61℃ 30SEC
72℃ 10SEC
22个循环 98℃ 10SEC
72℃ 10SEC
最终延伸 72℃ 2分钟
保持 12℃
实施例5：结果
对含有NNNNATGC PAM、ATNNNGC PAM和ATGCNNNN PAM的三个靶池进行筛选。图4显示了如体外测量的，MAD2001在含有NNNNATGC 3’PAM的靶上的耗尽图；图5显示了如体外测量的，MAD2007在含有NNNNATGC 3’PAM(上)和ATNNNNGC 3’PAM(下)的靶上的耗尽图；图6显示了如体外测量的，MAD2008在含有NNNNATGC3’PAM的靶上的耗尽图；图7显示了如体外测量的，MAD2008在含有NNNNATGC 3’PAM的靶上的耗尽图；图8显示了如体外测量的，MAD2011在含有NNNNATGC 3’PAM的靶上的耗尽图；并且图9显示了由以序列标志的形式显示的MAD裂解的靶的富集，总结了在体外筛选中有活性的挖掘的MAD系列RGN的PAM。因此，如在图4-图9观察到的，MAD2001、MAD2007、MAD2008、MAD2009和MAD2011核酸酶是有活性的。MAD2001切割包含NNRC PAM的靶，MAD2007偏好NNNSR PAM，MAD2008和MAD2009耗尽了具有NNAA PAM的靶。对于人类基因组编辑，PAM NNNNATGC、PAM ATNNNGC和PAM ATGCNNNN的限制性明显小于NGG或TTTV PAM(参见图11)。最后，类似于SPCas9，MAD2011具有NGG PAM。鉴定出最具活性的gRNA子池。由于每个gRNA子池包含64种不同的gRNA组合，我们在最具活性或最短的子池中测试每个gRNA以鉴定最佳的gRNA序列。鉴定了在体外测定中显示出最高的耗尽的三个gRNA。这些序列列在表6中。
表6
Figure BDA0003688304840000301
Figure BDA0003688304840000311
使用PAM信息和前三个gRNA设计，在HEK293T细胞中测试了MAD2001、MAD2007、MAD2008、MAD2009和MAD2011的活性。用核酸酶质粒和gRNA质粒共转染细胞。核酸酶由强CAG启动子表达，而gRNA由U6启动子表达。通过T7E1测定分析了细胞的插入缺失(indel)。如图10所示，五个MAD中有四个表现出活性。MAD2001对多于一个内源性靶具有活性，在含有GAGCPAM的靶上显示高达20％的插入缺失。这是第一次展示在哺乳动物细胞中有活性的古细菌CRISPR核酸酶。MAD2007、MAD2008和MAD2011对测试的靶显示出较低和可变的插入缺失水平。图11展示了新发现的精确编辑核酸酶的人类基因组覆盖范围，提供了人类细胞中精确编辑的覆盖范围。
此外，还鉴定了MAD系列核酸酶的切口酶和核酸酶失活变体，即MAD2001、MAD2007、MAD2008、MAD2009和MAD2011，其中这些切口酶和失活变体用于各种基于切口酶的精确编辑应用。下表7列出了切口酶和核酸酶失活变体的序列，其中与野生型MAD2001[SEQ ID No.1]核酸酶氨基酸序列(对于SEQ ID No.14-16)和MAD2007[SEQ ID No.7]核酸酶氨基酸序列(对于SEQ ID No.17-19)不同的氨基酸残基被突出显示和加粗。
表7
Figure BDA0003688304840000321
Figure BDA0003688304840000331
Figure BDA0003688304840000341
Figure BDA0003688304840000351
Figure BDA0003688304840000361
Figure BDA0003688304840000371
Figure BDA0003688304840000381
Figure BDA0003688304840000391
Figure BDA0003688304840000401
Figure BDA0003688304840000411
Figure BDA0003688304840000421
实施例6：通过测量GFP功能损失筛选在HEK293T-GFP细胞中有活性的MAD2001和 MAD2007间隔区：
为了测试MAD2007和MAD2001在哺乳动物细胞中是否有活性，设计了靶向HEK293t-GFP细胞中GFP基因座的间隔区文库。对于MAD2001，在两种gRNA支架(g1和g3)中设计了23个靶向NNRC PAM的间隔区。对于MAD2007，在3种gRNA支架(g1、g2和g3)中设计了43个靶向NNNSR PAM的间隔区。将gRNA克隆到pComplete质粒(CMV-MAD200x-U6-gRNA)并转化到大肠杆菌(NEB5α株)中。将所得的菌落挑入96孔的中孔板中并培养过夜。将大肠杆菌培养物用作PCR模板，扩增7kb的含有CMV启动子驱动的MAD2007/MAD2001和U6驱动的GRNA的片段。用150ng的未纯化PCR反应物和1μL的PolyFect转染试剂在96孔中转染20,000个HEK293T-GFP细胞。然后将细胞在37℃孵育96小时，然后进行流式细胞术。用流式细胞术测量各孔中的GFP-细胞和GFP+细胞。将每个孔中GFP-细胞的百分比绘图(参见图7)。相对于阴性对照，发现MAD2001和MAD2007的很多gRNA间隔区是有功能的。总体而言，在为MAD2001设计的23个间隔区中，有6个是有功能的。而为MAD2007设计的34/43间隔区是有功能的。使用这一GFP筛选鉴定的间隔区和PAM序列在表8示出。
表8
Figure BDA0003688304840000431
Figure BDA0003688304840000441
此外，在HEK293T细胞中有功能的34个MAD2007间隔区的PAM区域被用来生成序列标志，如图8所示。结果表明，在HEK293T细胞中MAD2007的PAM为NNNSR。
接下来，用质粒转染在HEK293T-GFP细胞中测试筛选中鉴定的一些MAD2007间隔区命中来验证。结果在图9中示出。相对于无gRNA对照，发现命中是有活性的。然而，取决于间隔区序列，有不同的活性水平。MAD2007的活性与MAD7相当，但低于SpCas9。
实施例7：MAD2007的密码子优化
到目前为止讨论的所有数据都是使用大肠杆菌密码子优化的MAD2007产生。对MAD2007进行优化以用于人类细胞优化，并且设计了两个密码子优化版本(hsMAD2007v1[SEQ ID NO.140]和hsMAD2007v2[SEQ ID No.141])。然后将设计物克隆到含有CMV驱动的MAD2007-T2A-dsRed和U6驱动的gRNA(g11，具有上文的支架1)的pComplete载体。对于每一个设计，挑取4个单独的克隆(c1、c2、c3和c4)，并在HEK293T-GFP细胞中测试GFP功能丧失。结果在图9中示出。基于dsRed表达，hsMAD2007v2相对于EcMAD2007显示更高的表达。此外，基于GFP-的百分比，hsMAD2007v2相对于EcMAD2007显示更高的活性。
实施例8：MAD2007同源物
在公共数据库中进行了MAD2007样蛋白序列的生物信息学搜索，并且鉴定出与MAD2007≥98％相同的来自阿扎夏普氏菌的三种序列。这些序列在表9中示出。
表9
Figure BDA0003688304840000451
Figure BDA0003688304840000461
Figure BDA0003688304840000471
虽然本发明通过许多不同形式的实施方案来满足，但是如结合本发明的优选的实施方案详细描述的，应理解本公开内容应被认为是对本发明的原理的示例而不意在将本发明局限于本文说明和描述的具体实施方案。本领域的技术人员可以作出许多变化而不脱离本发明的精神。本发明的范围将通过附加的权利要求和它们的等同物判断。摘要和标题不应被解释为限制本发明的范围，因为它们的目的是使适当的机构以及一般公众能够迅速确定本发明的一般性质。在以下的权利要求书中，除非使用术语“手段(means)”，否则其中列举的特征或要素都不应该被解释为根据35U.S.C.§112,
Figure BDA0003688304840000472
的手段加功能的限定。
序列表
<110> 因思科瑞普特公司
<120> 新颖的MAD核酸酶
<130> INSC036PCT
<140> PCT/US20/26095
<141> 2020-04-01
<150> 62/946,282
<151> 2019-12-10
<160> 144
<170> PatentIn version 3.5
<210> 1
<211> 1322
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 1
Met Lys Asn Thr Asn Glu Asp Tyr Tyr Leu Gly Leu Asp Ile Gly Thr
1 5 10 15
Asp Ser Val Gly Trp Ala Val Thr Asp Lys Glu Tyr Asn Ile Leu Glu
20 25 30
Phe Arg Arg Lys Pro Met Trp Gly Ile His Leu Phe Glu Gly Gly Ser
35 40 45
Thr Ala Gln Lys Thr Arg Val Tyr Arg Thr Ser Arg Arg Arg Leu Lys
50 55 60
Arg Arg Ala Glu Arg Ile Ala Leu Leu Arg Asp Ile Phe Ser Glu Glu
65 70 75 80
Ile Gly Lys Val Asp Pro Gly Phe Phe Glu Arg Leu Asp Glu Ser Asp
85 90 95
Leu His Leu Glu Asp Arg Val Thr Ser Gln Lys Asn Ser Leu Phe Asp
100 105 110
Asp Pro Glu Phe Asn Asp Lys Asp Leu His Lys Arg Phe Pro Thr Ile
115 120 125
Tyr His Leu Arg Arg His Leu Met His Ser Asn Arg Lys Glu Asp Ile
130 135 140
Arg Leu Ile Tyr Leu Ala Ala His His Ile Ile Lys Phe Arg Gly His
145 150 155 160
Phe Leu Tyr Lys Gly Ile Gly Asp Glu Glu Ile Pro Ser Phe Glu Ile
165 170 175
Val Leu Asn Ser Leu Ile Asp Asn Leu Arg Asp Glu Tyr Gly Met Glu
180 185 190
Leu Glu Val Ser Asp Arg Asp Leu Val Lys Ala Leu Leu Ser Asp Phe
195 200 205
Ser Ile Gly Ile Arg Glu Lys Ser Arg Glu Leu Ser Ser Cys Leu Asn
210 215 220
Ala Glu Ser Glu Asn Glu Lys Ala Leu Val Asp Phe Ile Ser Gly Lys
225 230 235 240
Lys Thr Asn Met Lys Lys Leu Phe Asp Asp Glu Ala Leu Asp Lys Met
245 250 255
Ser Phe Ser Leu Arg Asp Ser Gly Phe Glu Asp Gln Leu Arg Glu Asn
260 265 270
Glu Gly Val Leu Gly Pro Glu Arg Val His Thr Leu Glu Leu Ser Arg
275 280 285
Gln Ile Phe Glu Trp Ala Arg Leu Ser Ser Ile Leu Lys Asp Ser Asp
290 295 300
Ser Ile Ser Glu Ala Lys Ile Lys Asp Tyr Asp Gln His Arg Glu Asp
305 310 315 320
Leu Arg Met Leu Lys Arg Ala Val Lys Lys Tyr Ala Pro Asp Lys Tyr
325 330 335
Ser Glu Val Phe Lys Ser Lys Glu His Thr Gly Asn Tyr Cys Ser Tyr
340 345 350
Val Tyr Val Cys Gly Lys Gly Leu Pro Asp Lys Lys Cys Ser Thr Glu
355 360 365
Glu Phe Gln Lys Tyr Leu Lys Lys Ile Leu Asp Asp Ser Gly Val Arg
370 375 380
Asp Asp Glu Glu Phe Lys Thr Leu Ile Gln Arg Leu Asp Ala Gly Ile
385 390 395 400
Leu Cys Pro Lys Gln Arg Thr Gly Glu Asn Ser Val Ile Pro Tyr Ser
405 410 415
Val His Arg Lys Glu Leu Ile Gly Ile Leu Asn Asn Ala Ala Glu His
420 425 430
Tyr Pro Ser Leu Ser Arg Lys Gly Glu Asp Gly Phe Ser Ser Ile Asp
435 440 445
Lys Ile Leu Met Leu Glu Glu Phe Arg Ile Pro Tyr Tyr Val Gly Pro
450 455 460
Leu Asp Asp Arg Ser Ser Arg Ser Trp Leu Ile Arg Asn Ser Phe Glu
465 470 475 480
Ala Ile Thr Pro Trp Asn Phe Asn Glu Ile Val Asp Glu Asp Glu Thr
485 490 495
Ser Glu Arg Phe Ile Gly Asn Leu Thr Ser Met Cys Thr Tyr Leu Gly
500 505 510
Gly Glu Lys Val Leu Pro Lys Asn Ser Leu Leu Tyr Ser Arg Phe Met
515 520 525
Leu Tyr Asn Glu Ile Asn Asn Leu Arg Val Gly Gly Glu Lys Ile Pro
530 535 540
Ala Ala Leu Lys Asn Lys Met Val Ser Glu Leu Phe Ala Asn Arg Ala
545 550 555 560
Thr Ser Ser Lys Val Thr Leu Lys Glu Leu Lys Ala Phe Leu Lys Gly
565 570 575
Glu Gly Val Leu Thr Asp Ala Asp Glu Ile Ser Gly Ile Asp Asp Gly
580 585 590
Val Lys Ser Thr Leu Arg Ser Glu Ile Leu Ile Arg Lys Ile Ile Gly
595 600 605
Asp Lys Ile Ser Asp Arg Glu Met Ala Glu Glu Ile Val Arg Ile Leu
610 615 620
Thr Val Phe Gly Asp Glu Arg Arg Arg Ser Lys Ala Lys Leu Lys Lys
625 630 635 640
Glu Phe Ser Asp Lys Leu Thr Glu Lys Glu Ile Glu Lys Leu Ser Ser
645 650 655
Leu Lys Phe Asp Gly Trp Gly Arg Leu Ser Glu Lys Phe Leu Thr Gly
660 665 670
Leu Arg Gln Glu Val Asn Gly Arg Ser Met Ser Ile Ile Glu Ile Leu
675 680 685
Glu Asp Thr Asn Tyr Asn Leu Gln Glu Thr Leu Ser Lys Tyr Ser Phe
690 695 700
Asn Glu Ile Ile Asp Ser Tyr Asn Glu Val Leu Thr Ser Gly Pro Arg
705 710 715 720
Ser Ile Ser Tyr Asp Ile Leu Lys Asp Ser Tyr Leu Ser Pro Ala Val
725 730 735
Lys Arg Gly Val Trp Arg Ala Leu Ser Val Val Lys Asp Ile Leu Lys
740 745 750
Ala Val Gly Arg Pro Pro Lys Lys Ile Phe Val Glu Thr Thr Arg Glu
755 760 765
Glu Arg Glu Lys Lys Arg Thr Glu Ser Arg Lys Asp Ala Leu Met Tyr
770 775 780
Leu Tyr Lys Ser Cys Lys Glu Thr Glu Trp Glu Lys Arg Leu Asp Ser
785 790 795 800
Val Glu Glu Ser Ser Leu Arg Asn Arg Ser Leu Tyr Leu Tyr Tyr Thr
805 810 815
Gln Leu Gly Lys Cys Met Tyr Cys Gly Lys Asn Ile Asp Ile Gly Glu
820 825 830
Leu Asn Thr Asp Leu Ala Asp Arg Asp His Ile Tyr Pro Gln Ser Lys
835 840 845
Thr Lys Asp Asp Ser Ile Arg Asn Asn Leu Val Leu Val Cys Arg Gly
850 855 860
Cys Asn Gln Ala Lys Gly Asp Arg Tyr Pro Leu Pro Gln Glu Trp Val
865 870 875 880
Ser Arg Met His Ala Phe Trp Thr Met Leu Lys Asp Lys Gly Tyr Ile
885 890 895
Ser Ser Glu Lys Tyr Arg Arg Leu Thr Arg Arg Gly Glu Leu Thr Glu
900 905 910
Glu Glu Phe Gly Ala Phe Ile Asn Arg Gln Leu Val Glu Thr Ser Gln
915 920 925
Ser Ala Lys Ala Val Ile Thr Val Leu Lys Asn Ala Phe Lys Asp Ser
930 935 940
Asp Ile Val Tyr Val Lys Gly Ser Asn Val Ser Asp Phe Arg Ser Ser
945 950 955 960
Tyr Asn Phe Ile Lys Cys Arg Ser Val Asn Asp Tyr His His Ala Lys
965 970 975
Asp Ala Tyr Leu Asn Ile Val Val Gly Asn Val Leu Asp Thr Lys Phe
980 985 990
Thr Lys Asn Pro Ser Tyr Val Leu Lys Asn Arg Glu Gln Tyr Asn Ile
995 1000 1005
Gly Arg Met Tyr Asp Arg Asn Val Ser Arg Phe Gly Val Asp Ala
1010 1015 1020
Trp Val Ala Gly Asp Arg Gly Ser Ile Ala Thr Val Arg Lys Tyr
1025 1030 1035
Met Arg Arg Asn Asn Ile Leu Phe Thr Arg Tyr Ala Thr Lys Ser
1040 1045 1050
Lys Gly Ala Leu Phe Lys Glu Thr Val His Arg Lys Lys Glu Gly
1055 1060 1065
Leu Phe Glu Arg Lys Lys Gly Leu Glu Thr Glu Lys Tyr Gly Gly
1070 1075 1080
Tyr Ser Asp Ile Ser Thr Ser Tyr Leu Thr Leu Leu Glu Tyr Asp
1085 1090 1095
Lys Gly Lys Lys Arg Ile Arg Ser Leu Glu Ile Val Pro Thr Tyr
1100 1105 1110
Phe Ala Asn Thr Arg Pro Lys Glu Glu Asp Val Ile Arg Phe Phe
1115 1120 1125
Ser Glu Thr Arg Gly Leu Ala Asn Val Arg Val Val Met Pro Glu
1130 1135 1140
Val Arg Met Lys Ser Leu Phe Glu Tyr Arg Gly Phe Arg Phe His
1145 1150 1155
Val Thr Gly Ser Asn Gly Lys Gly Arg Phe Trp Ile Ser Ser Ala
1160 1165 1170
Ile Gln Leu Leu Leu Pro Glu Asn Leu Tyr Ala Tyr Cys Lys Ser
1175 1180 1185
Ile Glu Asn Asn Glu Lys Asp Ser Gln Arg Arg Ser Glu Lys Pro
1190 1195 1200
Leu Gln Asn Tyr Gly Phe Ser Ser Glu Met Asn Ile Glu Leu Phe
1205 1210 1215
Lys Cys Leu Met Asp Lys Ala Ala Lys Pro Pro Tyr Asp Val Lys
1220 1225 1230
Leu Ser Thr Leu Ser Lys Asn Leu Glu Glu Gly Phe Glu Lys Phe
1235 1240 1245
Lys Ala Leu Glu Leu Gly Pro Gln Val Lys Val Leu Gln Gln Ile
1250 1255 1260
Leu Asp Ile Tyr Ser Cys Asp Arg Lys Ser Gly Asp Leu Ser Val
1265 1270 1275
Leu Gly Ser Ala Arg Asn Ala Gly Arg Leu Asp Met Asn Gly Val
1280 1285 1290
Leu Ser Glu Ala Asp Gly Glu Gln Val Thr Met Ile Cys Gln Ser
1295 1300 1305
Pro Ser Gly Leu Phe Glu Lys Arg Val Pro Met Asn Glu Lys
1310 1315 1320
<210> 2
<211> 997
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 2
Met Lys Lys Arg Ile Phe Gly Phe Asp Ile Gly Ile Ala Ser Leu Gly
1 5 10 15
Trp Ala Val Val Asp Phe Asp Asp Thr Ala Asp Pro Glu Asn Asp Ile
20 25 30
Tyr Pro Thr Gly Glu Ile Val Lys Ser Gly Val Arg Cys Phe Pro Val
35 40 45
Ala Glu Asn Pro Lys Asp Gly Ser Ser Leu Ala Gln Pro Arg Arg Gln
50 55 60
Lys Arg Leu Leu Arg Arg Leu Cys Arg Arg Lys Ala Arg Arg Met Ala
65 70 75 80
Gly Ile Lys Asn Leu Phe Val Ala Asn Gly Leu Ile Gly Lys Asp Ala
85 90 95
Leu Phe Asn Glu Lys Ser Asn Ile Tyr Lys Ala Arg Asp Asn Ala Asp
100 105 110
Val Trp Asp Leu Arg Val Lys Ala Leu Thr Asp Lys Leu Thr Thr Ile
115 120 125
Glu Phe Ile Arg Val Leu Thr His Leu Ala Lys His Arg Gly Phe Lys
130 135 140
Ser Tyr Arg Ile Ala Ala Glu Lys Ala Asp Ala Glu Ser Gly Lys Val
145 150 155 160
Leu Glu Ala Val Lys Ala Asn Arg Ala Leu Leu Glu Asn Gly Lys Thr
165 170 175
Leu Ala Gln Ile Ile Val Glu Lys Gly Gly Gln Lys Arg Asn Arg Glu
180 185 190
Lys Met Ile Val Lys Asn Gly Lys Thr Glu Lys Gln Ala Ser Tyr Glu
195 200 205
Asn Ser Ile Pro Arg Asp Glu Ile Glu Arg Glu Thr Arg Leu Ile Phe
210 215 220
Glu Lys Gln Arg Ala Phe Gly Leu Glu Ala Ala Asn Glu Lys Leu Gln
225 230 235 240
Arg Asp Phe Glu Lys Ile Ala Phe Arg Phe Arg Glu Ile Lys Ser Lys
245 250 255
Asn Ile Glu Lys Met Ile Gly Lys Cys Glu Phe Glu Lys Asp Glu Pro
260 265 270
Arg Ala Pro Lys Asn Ala Pro Ser Ala Glu Phe Phe Val Ala Trp Thr
275 280 285
Lys Ile Asn Asn Cys Arg Val Arg Glu Pro Asp Gly Lys Ile Arg Phe
290 295 300
Leu Thr Gln Glu Glu Lys Glu Asn Val Phe Asn Leu Leu Lys Asp Gln
305 310 315 320
Lys Glu Val Lys Tyr Ser Ala Leu Lys Lys Ala Leu Phe Ala Lys Arg
325 330 335
Pro Asp Val Gln Phe Thr Asp Ile Glu Tyr Asn Pro Lys Pro Val Tyr
340 345 350
Asp Lys Lys Thr Gly Glu Ile Ile Glu Lys Thr Glu Asn Pro Glu Asn
355 360 365
Gln Lys Phe Phe Ser Leu Lys Gly Trp His Asp Leu Lys Ser Val Ile
370 375 380
Asp Val Ser Ser Tyr Pro Val Glu Thr Leu Asp Lys Ile Ala Thr Val
385 390 395 400
Ile Ala Thr Lys Lys Asn Asp Thr Asp Ile Ala Lys Gly Leu Lys Glu
405 410 415
Leu Asn Leu Pro Asp Ala Glu Ile Glu Lys Leu Thr Ser Leu Ser Phe
420 425 430
Ser Lys Phe Ile Arg Leu Ser Leu Lys Ala Leu Tyr Lys Ile Leu Pro
435 440 445
Glu Met Gln Lys Gly Met Lys Tyr Asn Glu Ala Cys Asp Ala Val Gly
450 455 460
Tyr Asp Phe Lys Ser Thr Gly Glu Ser Phe Ala Ala Gln Lys Gly Lys
465 470 475 480
Phe Leu Pro Pro Ile Pro Glu Ala Leu Ala Thr Thr Val Pro Val Val
485 490 495
Asn Arg Ala Met Thr Gln Phe Arg Lys Val Tyr Asn Ala Leu Ala Arg
500 505 510
Glu Tyr Gly Thr Pro Asp Gln Ile Asn Ile Glu Leu Ala Arg Asp Val
515 520 525
Tyr Asn Thr His Asp Glu Arg Lys Lys Ile Ala Asp Lys Gln Lys Glu
530 535 540
Tyr Gly Glu Glu Arg Lys Lys Ala Arg Asp Leu Ala Gln Glu Lys Met
545 550 555 560
Glu Ile Glu Asn Ile Ser Gly Arg Asp Leu Leu Lys Phe Arg Leu Tyr
565 570 575
Glu Gln Gln Asp Gly Lys Cys Ile Tyr Ser Gly Glu Thr Leu Asp Leu
580 585 590
Arg Arg Leu Thr Glu Gln Asp Tyr Cys Asp Val Asp His Ile Ile Pro
595 600 605
Tyr Ser Arg Ser Leu Asp Asn Ser Gln Asn Asn Lys Val Leu Cys Leu
610 615 620
Ser Arg Glu Asn Arg Arg Lys Ser Asp Lys Thr Pro Leu Glu Tyr Ile
625 630 635 640
Ile Asp Pro Val Lys Gln Ala Glu Phe Ile Ala Arg Val Lys Ser Met
645 650 655
Lys Gly Leu Ser Ala Pro Lys Arg Asp Arg Leu Leu Ile Arg Asp Phe
660 665 670
Lys Glu Lys Glu Leu Glu Phe Arg Asp Arg Asn Ile Asn Asp Thr Arg
675 680 685
Tyr Met Ala Arg Tyr Ile Met Lys Tyr Leu Asp Asp Cys Ile Asp Phe
690 695 700
Ser Gly Ser Gln Thr Asp Ile Lys Asp His Val Gln Ser Arg Ile Gly
705 710 715 720
Ser Leu Thr Asp Phe Leu Arg His Gln Trp Gly Leu His Lys Asp Arg
725 730 735
Asn Glu Asn Asp Arg His His Ala Gln Asp Ala Ile Val Ile Ala Cys
740 745 750
Ala Thr Asn Gly Tyr Thr Gln Tyr Leu Ala His Leu Ser Lys Ile Phe
755 760 765
Glu Asn Lys Gln Ala Tyr Ala Asn Lys Tyr Gly Gln Pro Trp Tyr Lys
770 775 780
Ala Phe Lys Gln His Val Lys Gln Pro Trp Asp Gly Phe Tyr Gln Asp
785 790 795 800
Val Gln Ala Ser Leu Ala Glu Ile Phe Val Ser Arg Pro Pro Arg Lys
805 810 815
Asn Ala Thr Gly Glu Val His Gln Asp Thr Ile Arg Thr Leu Asn Pro
820 825 830
Asn Lys Pro Gln Tyr Ser Glu Lys Asp Val Lys Ser Gly Ile Lys Leu
835 840 845
Arg Gly Gly Leu Ala Asn Asn Gly Asp Met Leu Arg Val Asp Val Phe
850 855 860
Ser Lys Lys Asn Ala Lys Gly Lys Glu Gln Phe Tyr Leu Val Pro Ile
865 870 875 880
Tyr Leu Ala Asp Arg Ile Lys Pro Glu Leu Pro Asn Lys Ala Ile Val
885 890 895
Ala Asn Lys Ser Glu Ser Glu Trp Ile Ile Met Asp Ala Thr Tyr Ser
900 905 910
Phe Lys Phe Ser Leu Tyr Met Asp Asp Leu Val Ser Val Ile Lys Gly
915 920 925
Asp Lys Lys Ile Phe Gly Tyr Tyr Lys Gly Thr Ser Arg Ser Thr Ala
930 935 940
Ser Ile Thr Ile Glu Gly His Asp Arg Asn Phe Ile Gln Pro Ser Ile
945 950 955 960
Gly Val Lys Thr Val Asp Asn Ile Lys Lys Tyr Gln Ile Asp Pro Leu
965 970 975
Gly Arg Tyr Val Glu Val Lys Ser Glu Ile Arg Leu Pro Leu Asn Ile
980 985 990
Lys Lys Arg Lys Ser
995
<210> 3
<211> 1392
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 3
Met Lys Lys Val Leu Gly Leu Asp Leu Gly Ser Ser Ser Ile Gly Trp
1 5 10 15
Ala Tyr Val His Glu Ala Glu Asn Glu Ala Glu Leu Gly Ser Ser Lys
20 25 30
Ile Ile Lys Leu Gly Val Arg Val Asn Pro Leu Thr Val Asp Glu Gln
35 40 45
Arg Asn Phe Glu Gln Gly Lys Ser Ile Thr Thr Asn Ala Ser Arg Thr
50 55 60
Leu Lys Arg Cys Met Arg Arg Asn Leu Gln Arg Tyr Lys Leu Arg Arg
65 70 75 80
Glu Asn Leu Ile Glu Val Leu Lys Lys His Gly Phe Ile Ser Asp Ala
85 90 95
Ser Ile Leu Ser Glu Gln Gly Asn Tyr Thr Thr Phe Glu Thr Tyr Arg
100 105 110
Leu Arg Ala Lys Ala Ala Val Ala Glu Ile Ser Leu Glu Glu Leu Ala
115 120 125
Arg Val Leu Leu Met Ile Asn Lys Lys Arg Gly Tyr Lys Ser Ser Arg
130 135 140
Lys Asn Arg Gly Gly Asp Glu Gly Lys Phe Ile Asp Gly Ile Ser Val
145 150 155 160
Ala Lys Gln Leu Tyr Asp Arg Gly Ile Thr Pro Gly Gln Phe Ser Leu
165 170 175
Glu Leu Leu Lys Glu Gly Lys Arg His Leu Pro Asp Tyr Tyr Arg Ser
180 185 190
Asp Leu Gln Asn Glu Leu Asp Arg Ile Trp Asn Phe Gln Gln Ser Phe
195 200 205
Tyr Pro Glu Ile Leu Thr Gln Asn Phe Arg Glu Gln Ile Arg Asp Lys
210 215 220
Gly Gln Lys Asn Thr Ser Gln Ile Phe Leu Arg Glu Tyr Gln Ile Tyr
225 230 235 240
Thr Ala Asp Asn Lys Gly Ala Asp Lys Leu Ser Arg Ala Leu Gln Trp
245 250 255
Arg Val Glu Gly Leu Ser Arg Lys Leu Ser Val Glu Glu Leu Ala Phe
260 265 270
Val Met Ser Asp Leu Asn Gly Ser Ile Ser Gly Ser Ser Gly Tyr Leu
275 280 285
Gly Ala Ile Gly Asp Arg Ser Lys Glu Leu Tyr Leu Gly Lys Gln Thr
290 295 300
Val Gly Gln Tyr Leu Met Glu Lys Leu Asn Thr Asn Pro Asn Gly Ser
305 310 315 320
Leu Lys Ser Lys Val Phe Tyr Arg Gln Asp Tyr Leu Asp Glu Phe Glu
325 330 335
Arg Ile Trp Glu Thr Gln Ala Gly Phe His Lys Glu Leu Thr Leu Glu
340 345 350
Leu Lys Lys Glu Ile Arg Asp Ile Ile Ile Phe Tyr Gln Arg Ser Leu
355 360 365
Lys Ser Gln Lys Gly Leu Ile Ser Phe Cys Glu Leu Glu Ser Lys Leu
370 375 380
Val Glu Ile Glu Val Asn Gly Lys Met Arg Arg Lys Val Val Gly Ser
385 390 395 400
Arg Val Cys Pro Lys Ser Ser Pro Leu Phe Gln Glu Phe Lys Ile Trp
405 410 415
Ser Ile Leu Asn Asn Ile Cys Val Trp Ser Val Asp Lys Ser Lys Ser
420 425 430
Ser Glu Ala Arg Lys Met Asp Glu Arg Asp Lys Glu Pro Asn Leu Asn
435 440 445
Gln Glu Glu Lys Glu Ile Leu Phe Lys Glu Leu Ser Leu Lys Glu Lys
450 455 460
Leu Ser Lys Arg Asp Val Leu Glu Leu Leu Phe Glu Asp Ala Arg Lys
465 470 475 480
Leu Asp Met Asn Tyr Glu Lys Val Glu Gly Asn Arg Thr Gln Ala Thr
485 490 495
Leu Phe Lys Ala Tyr Gln Glu Ile Ile Ala Arg Ser Gly His Gly Glu
500 505 510
Tyr Asp Phe Thr Arg Met Leu Ser Ser Glu Ile Leu Glu Ile Val Ser
515 520 525
Gly Val Phe Asp Gly Leu Gly Tyr Asn Thr Asp Ile Leu Tyr Phe Asn
530 535 540
Ser Glu Gly Glu Leu Asp Gln Gln Pro Leu Tyr Arg Leu Trp His Leu
545 550 555 560
Leu Tyr Ser Phe Glu Gly Asp Lys Ser Asn Ser Gly Asn Glu Asn Leu
565 570 575
Ile Asn Lys Ile Thr Asn Leu Tyr Gly Phe Asp Arg Glu Tyr Ala Val
580 585 590
Ile Leu Ala Asp Val Val Phe Pro Pro Asp Tyr Gly Asn Leu Ser Ala
595 600 605
Lys Ala Ile His Lys Ile Leu Pro Tyr Leu Lys Asp Gly Asn Lys Tyr
610 615 620
Ser Leu Ala Cys Glu Tyr Ala Gly Phe Arg His Ser Lys Asn Ser Leu
625 630 635 640
Thr Lys Glu Glu Arg Glu Lys Arg Val Leu Lys Glu Arg Leu Asp Ile
645 650 655
Leu Pro Lys Asn Thr Leu Arg Asn Pro Val Val Glu Lys Ile Leu Asn
660 665 670
Gln Met Val His Val Val Asn Gly Val Ile Asn Lys Tyr Gly Lys Pro
675 680 685
Asp Glu Ile Arg Ile Glu Leu Ala Arg Glu Leu Lys Lys Asn Ala Lys
690 695 700
Glu Arg Glu Glu Trp Thr Arg Ala Ile Asn Lys Ser Thr Ile Glu Asn
705 710 715 720
Glu Lys Leu Arg Ser Val Leu Lys Lys Glu Phe Gly Phe Thr Gln Val
725 730 735
Ser Arg Asn Asp Ile Val Arg Tyr Lys Leu Tyr Leu Glu Leu Glu Ser
740 745 750
Arg Gly Phe Lys Thr Leu Tyr Ser Asn Thr Tyr Ile Pro Leu Glu Lys
755 760 765
Leu Phe Phe Lys Glu Phe Asp Ile Glu His Ile Ile Pro Gln Ser Arg
770 775 780
Leu Phe Asp Asp Ser Phe Ser Asn Lys Thr Ile Glu Leu Arg Ser Val
785 790 795 800
Asn Gln Glu Lys Asp Asn Gln Thr Ala Tyr Asp Tyr Val Ser Gly Lys
805 810 815
Gly Gly Glu Ala Gly Leu Gln Glu Tyr Leu Glu Arg Val Glu Asp Leu
820 825 830
Phe Lys Gly Gly Tyr Ile Asn Lys Ala Lys Tyr Asn Lys Leu Arg Met
835 840 845
Thr Gly Lys Asp Ile Pro Asp Asp Phe Ile Asp Arg Asp Leu Arg Asp
850 855 860
Thr Gln Tyr Ile Ala Arg Arg Ala Lys Ala Met Leu Glu Glu Val Val
865 870 875 880
Gly Asn Val Val Ser Thr Ser Gly Ala Val Thr Asp Arg Leu Arg Glu
885 890 895
Asp Trp Gln Leu Val Asp Val Met Lys Glu Leu Asn Trp Asn Lys Tyr
900 905 910
Glu Arg Leu Gly Leu Thr Glu Ile Val Glu Asp Arg Asp Gly Arg Lys
915 920 925
Ile Arg Arg Ile Lys Gly Trp Thr Lys Arg Asn Asp His Arg His His
930 935 940
Ala Met Asp Ala Leu Thr Ile Ala Phe Thr Lys Pro Lys Tyr Val Gln
945 950 955 960
Tyr Leu Asn Asn Leu Asn Ala Arg Gly Asp Lys Ser Ser Ser Val Tyr
965 970 975
Gly Ile Glu Arg Asp Glu Leu Ser Arg Asp Ser Lys Gly Lys Leu Arg
980 985 990
Phe Asn Ser Pro Met Pro Leu Lys Glu Phe Arg Met Glu Ala Lys Leu
995 1000 1005
His Leu Glu Asn Val Leu Val Ser Thr Lys Ala Lys Asn Lys Val
1010 1015 1020
Ile Thr Pro Asn Val Asn Lys Ser Lys Lys Arg Gly Gly Met Asn
1025 1030 1035
Gln Lys Val Gln Leu Thr Pro Arg Gly Gln Leu His Gln Glu Thr
1040 1045 1050
Ile Tyr Gly Ser Ile Lys Gln Tyr Val Thr Lys Glu Val Lys Val
1055 1060 1065
Gly Ser Ala Phe Asn Met Glu Met Ile Leu Lys Val Ala Asn Lys
1070 1075 1080
Ala Tyr Arg Glu Ala Leu Leu Lys Arg Leu Asn Ala Phe Asp Gln
1085 1090 1095
Asp Ala Lys Lys Ala Phe Thr Gly Lys Asn Ser Leu Glu Lys Asn
1100 1105 1110
Pro Ile Phe Ile Asn Asp Ser His Thr Cys Lys Val Pro Glu Lys
1115 1120 1125
Val Lys Val Val Ser Phe Glu Thr Val Tyr Thr Ile Arg Lys Glu
1130 1135 1140
Ile Gly Pro Asp Leu Lys Val Asp Lys Val Ile Asp Lys Arg Val
1145 1150 1155
Arg Asp Ile Leu Glu Thr Arg Leu Val Glu Phe Gly Gly Asp Ser
1160 1165 1170
Lys Leu Ala Phe Thr Asn Leu Asp Glu Asn Pro Ile Trp Leu Asn
1175 1180 1185
Lys Glu Lys Gly Ile Asp Ile Lys Arg Val Thr Ile Ser Gly Val
1190 1195 1200
Ser Asn Val Ile Ala Leu His Asp Lys Leu Asp Lys Asp Gly Lys
1205 1210 1215
Leu Val Leu Asp Glu His Gly Gln Pro Gln Pro Val Asp Phe Val
1220 1225 1230
Cys Ser Gly Asn Asn His His Val Val Val Tyr Arg Asp Pro Glu
1235 1240 1245
Gly Lys Ile Gln Asp Asp Val Val Ser Phe Phe Glu Ala Thr Val
1250 1255 1260
Arg Ala Lys Glu Gly Leu Pro Val Ile Asp Arg Glu Tyr Lys Lys
1265 1270 1275
Gln Glu Gly Trp Glu Phe Leu Phe Ser Met Lys Gln Asn Glu Tyr
1280 1285 1290
Phe Val Phe Pro Asn Glu Glu Thr Gly Phe Asp Pro Lys Glu Val
1295 1300 1305
Asp Leu Met Asn Pro Asp Asn Tyr Ala Leu Ile Ser Pro Asn Leu
1310 1315 1320
Phe Arg Val Gln Thr Met Ser Arg Val Met Tyr Gly Asn Gln Val
1325 1330 1335
Val Arg Asp Tyr Lys Phe Arg His His Leu Glu Thr Thr Val Lys
1340 1345 1350
Asp Cys Lys Glu Leu Lys Asp Ile Ala Tyr Lys Gln Tyr Lys Ser
1355 1360 1365
Leu Asp Phe Ala Ser Gln Ile Val Lys Val Arg Ile Asp His Ile
1370 1375 1380
Gly Gln Ile Val His Val Gly Glu Tyr
1385 1390
<210> 4
<211> 1133
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 4
Met Ala Asp Lys Glu Lys Leu Lys Glu Thr Tyr Thr Ile Gly Leu Asp
1 5 10 15
Ile Gly Ile Ala Ser Val Gly Trp Ala Ile Leu Gly Glu Asn Arg Ile
20 25 30
Ile Asp Leu Gly Val Arg Cys Phe Asp Lys Ala Glu Thr Ala Lys Glu
35 40 45
Gly Glu Ser Leu Asn Leu Ser Arg Arg Met Ala Arg Leu Met Arg Arg
50 55 60
Arg Leu Arg Arg Arg Ala Trp Arg Leu Thr Lys Leu Ala Arg Leu Leu
65 70 75 80
Lys Arg Val Gly Leu Ile Ala Asp Val Gly Val Leu Lys Gln Pro Pro
85 90 95
Ser Lys Gly Phe Gln Thr Pro Asn Leu Trp Gln Leu Arg Val Glu Gly
100 105 110
Leu Asp Arg Lys Leu Gly Asp Asp Glu Trp Ala Arg Val Ile Tyr His
115 120 125
Leu Cys Lys His Arg Gly Phe His Trp Ile Ser Lys Ala Glu Ala Lys
130 135 140
Ala Ala Asp Ser Asp Lys Glu Gly Gly Lys Val Lys Gln Gly Leu Ala
145 150 155 160
Gly Thr Lys Arg Leu Met Gln Glu Lys Ser Tyr Arg Thr Ala Ala Glu
165 170 175
Met Val Leu Ala Glu Phe Pro Asp Ala Gln Arg Asn Lys Gln Gly Glu
180 185 190
Tyr Ser Lys Ala Leu Ser Arg Glu Leu Leu Cys Glu Glu Leu Lys Glu
195 200 205
Leu Phe Lys Gln Gln Arg Ala Phe Gly His Thr His Ala Asp Asp Lys
210 215 220
Leu Glu Thr Asn Ile Leu Gly Asn Gly Asp Lys Lys Ser Gly Leu Phe
225 230 235 240
Trp Val Gln Lys Pro Ser Leu Ser Gly Glu Ala Leu Leu Lys Met Leu
245 250 255
Gly Lys Cys Thr Phe Glu Lys Asp Glu Tyr Arg Ala Ala Lys Ala Cys
260 265 270
Phe Thr Ala Glu Arg His Val Leu Leu Thr Arg Ile Asn Asn Leu Arg
275 280 285
Ile Val Glu Asn Gly Lys Met Arg Gly Leu Thr Ala Asp Glu Arg Arg
290 295 300
Ile Ala Leu Trp Gln Pro Tyr Gln Gln Ala Gly Asp Phe Thr Phe Lys
305 310 315 320
Gln Leu Gly Ser Ala Leu Glu Lys His Gly Ser Leu Leu Lys Gly Gly
325 330 335
Tyr Lys Phe Ala Gly Leu Thr Tyr Pro Arg Glu Thr Asp Glu Lys Ala
340 345 350
Lys Asn Pro Glu Thr Ala Thr Leu Val Lys Ile Pro Ala Trp Gln Glu
355 360 365
Leu Lys Lys Thr Leu Ile Gly Val Gly Leu Glu Arg Glu Trp Gln Gly
370 375 380
Met Ala Asp Ala Ala Leu Asn Gly Lys Pro Asp Leu Leu Asp Lys Ile
385 390 395 400
Gly Trp Val Leu Ser Val Tyr Lys Glu Asp Asp Glu Val Glu Leu Glu
405 410 415
Leu Gly Lys Leu Gln Leu Ala Ala Asn Val Ile His Ala Leu Gln Thr
420 425 430
Val Arg Phe Asp Lys Phe Ser Asn Leu Ser Leu Leu Ala Leu Cys Lys
435 440 445
Ile Leu Pro Gln Met Glu Asn Gly Met Arg Tyr Asp Glu Ala Cys Lys
450 455 460
Glu Ala Gly Tyr Gln His Ser Met Pro Asp Met Gln Asp Leu Glu Ala
465 470 475 480
Lys Val Arg Tyr Leu Pro Pro Phe Tyr Ser Gly Arg Glu Lys Asp Gly
485 490 495
Arg Leu Lys Phe Asn Glu Asp Met Asp Ile Pro Arg Asn Pro Val Val
500 505 510
Leu Arg Ala Leu Asn Gln Ala Arg Lys Val Val Asn Ala Leu Ile Arg
515 520 525
Lys Tyr Gly Ser Pro His Ala Val His Ile Glu Met Ala Arg Asp Leu
530 535 540
Ser Arg Pro Ile Asp Glu Arg Arg Lys Ile Glu Arg Asp Gln Ala Asp
545 550 555 560
Tyr Arg Thr Lys Asn Glu Asp Ala Arg Lys Ala Phe Ala Ser Asp Phe
565 570 575
Gly Phe Glu Pro Lys Gly Arg Gln Phe Glu Lys Tyr Met Leu Tyr Arg
580 585 590
Glu Gln Gln Ala Lys Cys Ala Tyr Ser Leu Ala Pro Leu Asp Leu Asn
595 600 605
Arg Val Leu Asn Asp Gln Gly Tyr Ala Glu Val Asp His Ala Leu Pro
610 615 620
Tyr Ser Arg Ser Phe Asp Asp Gly Lys Asn Asn Arg Val Leu Val Leu
625 630 635 640
Thr Ser Glu Asn Arg Asn Lys Gly Asn Gln Thr Pro Tyr Glu Tyr Leu
645 650 655
Asp Gly Thr Ser Asp Ser Glu Lys Trp Arg Leu Phe Glu Ser Phe Val
660 665 670
Ser Gly Asn Lys Ala Tyr Ser Gln Ala Lys Arg Asn Arg Leu Leu Lys
675 680 685
Lys Asp Phe Gly Val Lys Glu Thr Gln Asp Phe Lys Glu Arg Asn Leu
690 695 700
Asn Asp Thr Arg Tyr Ile Cys Arg Phe Phe Lys Asn Tyr Val Glu Gln
705 710 715 720
Tyr Leu Gln Leu His Glu Glu Ser Asp Ala Lys Arg Cys Val Val Val
725 730 735
Ser Gly Gln Leu Thr Ser Leu Leu Arg Phe Arg Trp Gly Ile Asn Lys
740 745 750
Ile Arg Ser Glu Ser Asp Arg His His Ala Leu Asp Ala Ala Val Val
755 760 765
Ala Ala Cys Ser His Gly Leu Val Lys Arg Met Ser Asp Tyr Ser Arg
770 775 780
Arg Lys Glu Leu Gly Gln Val Arg Asp Arg Ile Glu Lys Val Asp Lys
785 790 795 800
Lys Thr Gly Glu Ile Ile Asp His Phe Pro Ser Pro Trp Ala His Phe
805 810 815
Arg Gln Glu Leu Leu Ala Arg Leu His Ile Asp Asp Ala Asn Glu Leu
820 825 830
Arg Ala Val Val Glu Asn Leu Gly Thr Tyr Pro Pro Glu Ala Leu Glu
835 840 845
Ser Leu Thr Pro Leu Phe Val Ser Arg Ala Pro Gln Arg Arg Asn Ser
850 855 860
Gly Ala Ala His Lys Glu Thr Ile Tyr Ala Gln Pro Glu Ala Met Lys
865 870 875 880
Glu Lys Gly Ser Val Thr Gln Lys Val Ala Val Thr Ser Leu Lys Pro
885 890 895
Ala Asp Val Asp Lys Leu Ile Asp Pro Glu Arg Asn Val Lys Leu Tyr
900 905 910
Ala Tyr Leu Arg Lys Trp Leu Ala Gly Lys Asp Glu Arg Glu Lys Arg
915 920 925
Ala Lys Ala Ile Glu Ala Ser Ala Gly Arg Gly Lys Glu Lys Arg Asp
930 935 940
Leu Thr Pro Glu Glu Lys Ile Glu Ile Glu Arg Leu Arg Ala Leu Pro
945 950 955 960
Gln Lys Pro Asp Lys Gln Gly Lys Pro Thr Gly Pro Ile Val Arg Ala
965 970 975
Val Thr Met Val Ile Asp Lys Leu Ser Gly Ile Pro Val Arg Gly Gly
980 985 990
Ile Ala Lys Asn Asp Thr Met Leu Arg Val Asp Met Phe Ser Lys Ala
995 1000 1005
Lys Arg Tyr Tyr Leu Val Thr Val Tyr Val Phe His Ser Val Ala
1010 1015 1020
Lys Glu Leu Pro Ser Arg Ala Ile Val Ala His Lys Asp Glu Asp
1025 1030 1035
Asp Trp Thr Val Ile Ser Glu Asp Phe Glu Phe Cys Phe Ser Met
1040 1045 1050
Tyr Pro Asn Asp Phe Ile Arg Ile Ser Gln Lys Lys Glu Thr Phe
1055 1060 1065
Met Gly Tyr Tyr Ala Gly Cys Asp Arg Gly Ser Gly Asn Val Asn
1070 1075 1080
Leu Trp Ser His Asp Arg Asn Ser Gln Ile Gly Lys Ser Gly Met
1085 1090 1095
Ile Arg Gly Ile Gly Val Lys Thr Ala Val Asn Val Glu Lys Phe
1100 1105 1110
Asn Val Asp Val Leu Gly Asn Ile Tyr Pro Ala Pro Pro Glu Ile
1115 1120 1125
Arg Arg Glu Leu Ala
1130
<210> 5
<211> 1049
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 5
Met Gly Tyr Ile Leu Gly Ile Asp Ile Gly Ile Ala Ser Ile Gly Phe
1 5 10 15
Ala Gly Val Asn His Asp Leu Lys Lys Ile Leu Phe Ser Gly Val His
20 25 30
Ile Phe Glu Ala Ala Glu Asn Pro Lys Thr Gly Ala Ser Leu Ala Glu
35 40 45
Pro Arg Arg Thr Ala Arg Gly Gln Arg Arg Val Ile His Arg Arg Ala
50 55 60
Gln Arg Lys Asn Ala Ile Arg Gln Leu Leu Leu Arg His Gly Leu Asn
65 70 75 80
Cys Leu Ser Val Val Asp Lys Lys Tyr Glu Pro Thr Gly Lys Asn Thr
85 90 95
Pro Pro Ile Ser Pro Trp Asp Leu Arg Arg Thr Ala Leu Asp Arg Lys
100 105 110
Leu Thr Asp Glu Glu Leu Val Arg Ile Leu Phe His Ile Gly Lys His
115 120 125
Arg Gly Phe Gln Ser Asn Lys Lys Ser Gln Ser Asn Glu Gly Asp Asp
130 135 140
Gly Lys Ala Leu Lys Gly Ala Gly Asp Leu Glu Gln Lys Trp Ile Gln
145 150 155 160
Ser Gly Glu Lys Thr Ile Gly Ala Tyr Leu Ser Thr Gln Ser Lys Lys
165 170 175
Arg Asn Gly Asn Glu Ser Tyr Asp Asn Phe Ile Lys Arg Asp Trp Leu
180 185 190
Arg Glu Glu Ile Lys Val Ile Phe Glu Ala Gln Arg Lys Phe Asn Gln
195 200 205
Ile Lys Ala Thr Glu Val Leu Arg Leu Glu Tyr Ala Gly Thr Gly Glu
210 215 220
Lys Ala Lys Arg Asn Thr Pro Glu Gly Asp Gly Ile Ala Phe Tyr Gln
225 230 235 240
Arg Pro Leu Gln Ser Ser Glu Lys Leu Ile Gly Asp Cys Thr Phe Glu
245 250 255
Lys Gly Glu Lys Arg Ala Pro Lys Phe Ser Tyr Thr Ala Glu Leu Phe
260 265 270
Val Leu Trp Ser Arg Leu Asn Asn Thr Lys Ile Lys Ile Gln Asn Gly
275 280 285
Asp Glu Arg Phe Leu Thr Gln Asp Glu Lys Asn Lys Leu Val Asn Leu
290 295 300
Ala His Lys Asn Lys Gly Gly Val Ser Tyr Thr Gln Ala Arg Lys Glu
305 310 315 320
Ile Gly Leu Asn Glu Ser Glu Arg Phe Asn Ile Ser Tyr Arg Gln Leu
325 330 335
Asp Lys Gly Asp Asn Ser Trp Glu Lys Ile Arg Asn Glu Ala Glu Lys
340 345 350
Ser Asn Phe Leu Lys Leu Ser Gly Phe His Ala Leu His Glu Ala Leu
355 360 365
Asp Thr Gly Ser Ala Thr Asp Trp Gln Lys Trp Ile Gly Ser Asp Arg
370 375 380
Asp Lys Leu Asp Glu Val Ala Tyr Ile Thr Ser Phe Ile Glu Asp Gly
385 390 395 400
Lys Ile Ile Arg Glu Lys Tyr Gln Lys Leu Gly Leu Asn Glu Asp Gln
405 410 415
Ile Lys Lys Leu Cys Glu Ile Lys Asn Phe Ser Lys Thr Val Asp Leu
420 425 430
Ser Leu Lys Ala Leu Arg Asn Ile Leu Pro Glu Leu Glu Lys Gly Leu
435 440 445
Arg Tyr Asp Glu Ala Cys Lys Ala Leu Asn Tyr Asn Asn Gln Pro Glu
450 455 460
Asn Lys Gly Leu Ser Lys Val Pro Lys Phe Glu Asp Val Arg Asn Pro
465 470 475 480
Val Val Asn Arg Ala Leu Gly Gln Thr Arg Lys Val Ile Asn Ala Cys
485 490 495
Ile Arg Glu Tyr Gly Leu Pro Asp Thr Ile Val Val Glu Leu Ala Arg
500 505 510
Glu Val Gly Lys Asn Phe Arg Asp Arg Lys Asp Ile Glu Lys Glu Gln
515 520 525
Lys Thr Asn Glu Ala Arg Arg Asn Thr Ala Lys Thr His Ile Ala Glu
530 535 540
Ile Leu Gly Ile Ile Glu Asp Asn Val Thr Gly Glu Asp Ile Leu Lys
545 550 555 560
Tyr Arg Leu Trp Lys Glu Gln Asp Cys Phe Cys Pro Tyr Ser Gly Ala
565 570 575
Tyr Ile Thr Pro Glu Met Leu Arg Asp Ser Thr Ser Val Gln Ile Asp
580 585 590
His Ile Ile Pro Tyr Ser Arg Ser Trp Asp Asn Ser Tyr Met Asn Lys
595 600 605
Val Leu Val Leu Thr Thr Glu Asn Gln Lys Lys Lys Asn Asp Thr Pro
610 615 620
Phe Glu Tyr Leu Gly Lys Thr Asn Arg Trp Glu Ala Leu Glu Val Phe
625 630 635 640
Ala Arg Gln Leu Pro Pro Lys Lys Ala Glu Arg Leu Leu Thr Glu Asn
645 650 655
Phe Asp Asp Lys Lys Ala Gly Glu Trp Lys Asp Arg Ala Leu Asn Asp
660 665 670
Thr Arg Tyr Met Ala Arg Leu Leu Lys Thr His Leu Glu Gln Ser Leu
675 680 685
Asp Leu Gly Lys Gly Asn Arg Val Gln Thr Arg Asn Gly Ser Leu Thr
690 695 700
Ala His Leu Arg Gly Ala Trp Gly Phe Pro Asp Lys Asn Arg Arg Asn
705 710 715 720
Asp Arg His His Ala Leu Asp Ala Ile Val Ile Ala Cys Ser Thr Gln
725 730 735
Ser Met Val Gln Gly Leu Thr Asn Trp Asn Lys Tyr Glu Ala Arg Arg
740 745 750
Lys Asn Pro Ala Glu Arg Pro Leu Pro Pro Lys Pro Trp Glu Ser Phe
755 760 765
Arg Glu Asp Ala Lys Glu Ser Val Asn Ser Ile Phe Val Ser Arg Met
770 775 780
Pro Val Arg Thr Ile Ser Gly Ala Ala His Glu Asp Thr Ile Arg Ser
785 790 795 800
Ile Arg Lys Ser Asp Gly Lys Ile Ile Gln Arg Ile Lys Leu Lys Asp
805 810 815
Phe Lys Lys Asp Thr Leu Glu Asn Met Val Asp Lys Ala Arg Asn Ile
820 825 830
Lys Leu Tyr Asp Ile Leu Lys Glu Arg Leu Asp Ala His Gly Gly Asp
835 840 845
Ala Lys Lys Ala Phe Ala Thr Pro Val Tyr Met Pro Val Asn Asp Pro
850 855 860
Ser Lys Pro Ala Pro Arg Ile Asn Ser Val Arg Ile Leu Thr Asn Glu
865 870 875 880
Lys Ser Gly Ile Glu Ile Asn His Gly Leu Ala Ser Asn Gly Asp Met
885 890 895
Val Arg Val Asp Val Phe Lys Lys Asp Asn Lys Phe Trp Leu Val Pro
900 905 910
Ile Tyr Val His His Phe Ala Glu Asp Lys Leu Pro Asn Lys Ala Ile
915 920 925
Met Gln Gly Lys Asp Glu Arg Glu Trp Glu Glu Met Asn Asp Asp Asp
930 935 940
Phe Met Phe Ser Leu Tyr Arg Asn Asp Leu Ile Lys Val Thr Thr Lys
945 950 955 960
Lys Glu Thr Met Leu Val Tyr Phe Gly Gly Leu Asp Arg Ala Thr Gly
965 970 975
Asn Ile Ser Ile Lys Ala His Asp Arg Asp Pro Ser Phe Gly Thr Asn
980 985 990
Gly Glu Asn Arg Thr Gly Val Lys Thr Ala Ile Asn Phe Glu Lys Phe
995 1000 1005
Ser Val Asn Tyr Phe Gly Arg Lys His Lys Ile Glu Lys Glu Lys
1010 1015 1020
Arg Leu Gly Val Ala His Ser Asp Asp Ser Glu Arg Gly Ala Ala
1025 1030 1035
Ile Pro Glu Gln Gly Thr Gly Ala Ala Ala Glu
1040 1045
<210> 6
<211> 1501
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 6
Met Lys Arg Ile Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Val Ile Asn Gln Asp Asn Ile Asn Asp Lys Asp Ile Leu Thr Gly
20 25 30
Ile Glu Cys Thr Gly Ser Arg Ile Ile Pro Met Asp Ala Ala Thr Leu
35 40 45
Gly Asp Phe Asp Arg Gly Asn Ala Gln Ser Gln Thr Ala Asp Arg Thr
50 55 60
Lys Arg Arg Ser Ala Arg Arg Leu Ile Glu Arg Ser His Ile Arg Arg
65 70 75 80
Glu Arg Leu Asn Arg Val Leu Met Thr Met Gly Trp Leu Pro Glu His
85 90 95
Tyr Ser Asp Ser Leu Asp Arg Tyr Gly Lys Leu Ser Lys Gly Thr Glu
100 105 110
Gln Lys Ile Ala Trp Lys Lys Ser Gly Asn Gly Asn Tyr Glu Phe Ile
115 120 125
Phe Lys Asp Ser Phe Asn Glu Met Leu Asp Asp Phe Lys Asn Glu His
130 135 140
Pro Asp Phe Ala Gln Arg Gly Leu Lys Ile Pro Tyr Asp Trp Thr Ile
145 150 155 160
Tyr Tyr Leu Arg Lys Lys Ala Leu Thr His Pro Val Thr Asn Gln Glu
165 170 175
Leu Ala Trp Ile Leu His Ser Phe Asn Gln Lys Arg Gly Tyr Tyr Gln
180 185 190
Arg Gly Glu Glu Glu Glu Gln Gln Pro Asp Lys Lys Ile Glu Tyr Ile
195 200 205
Pro Leu Lys Val Lys Glu Ile Arg Glu Thr Gly Glu Thr Lys Gly Ala
210 215 220
Asp Lys Trp Phe Glu Leu Ile Leu Glu Asn Asp Leu Val Tyr Lys Arg
225 230 235 240
Thr Phe Lys Glu Met Pro Asp Trp Lys Gly Lys Thr Leu Glu Leu Ile
245 250 255
Val Thr Thr Asp Leu Asp Lys Asp Gly Asn Pro Val Ile Lys Asp Gly
260 265 270
Lys Ala Lys Tyr Ser Ile Arg Ala Pro Lys Glu Asp Asp Trp Thr Leu
275 280 285
Val Lys Val Arg Thr Gln Ser Asp Ile Arg Lys Ser Gly Lys Thr Val
290 295 300
Gly Cys Tyr Ile Tyr Asp Ala Leu Ile Arg Lys Pro Asp Ile Lys Ile
305 310 315 320
Arg Gly Lys Leu Val Arg Thr Ile Glu Arg Glu Phe Tyr Arg Glu Glu
325 330 335
Leu Glu Gln Ile Leu Lys Lys Gln Lys Glu Phe Asn Gln Asp Leu Arg
340 345 350
Asp Lys Glu Leu Tyr Asn Glu Cys Ile Gly Val Leu Tyr Pro Asn Asn
355 360 365
Asp Thr His Arg Lys Glu Ile Ala Asn Arg Asp Asp Phe Ala Tyr Leu
370 375 380
Phe Ile Asn Asp Ile Ile Phe Tyr Gln Arg Pro Leu Lys Ser Lys Lys
385 390 395 400
Ser Leu Ile Ser Asp Cys Pro Tyr Glu Glu Arg Ile Tyr Lys Asp Lys
405 410 415
Ser Thr Gly Gln Lys Leu Thr Ser Ala Ile Lys Cys Ile Pro Lys Ser
420 425 430
His Pro Thr Tyr Gln Glu Phe Arg Leu Trp Gln Phe Leu Ser Tyr Leu
435 440 445
Lys Ile Tyr Glu Lys Glu Arg Thr Glu Ile Gly Lys Ile Gln Thr Asp
450 455 460
Ile Asp Ile Thr Asp Ile Leu Leu Pro Asp Asn Glu Ser Tyr Ala Ala
465 470 475 480
Leu Phe Lys Lys Leu Asn Asp Glu Ala Glu Ile Lys Gln Asp Lys Ile
485 490 495
Leu Lys Tyr Phe Pro Gln Leu Lys Lys Asn Ile Lys Asn Phe Arg Trp
500 505 510
Asn Tyr Pro Glu Asp Lys Thr Tyr Pro Gly Asn Thr Thr Arg Ala Glu
515 520 525
Met Leu Lys Arg Leu Lys Lys Ala Asn Ile Gly Ser Asp Phe Leu Thr
530 535 540
Thr Glu Gln Glu Thr Ala Leu Trp His Ile Leu Tyr Ser Val Asn Asp
545 550 555 560
Lys Ala Glu Leu Glu Lys Ala Leu Ser Thr Phe Ala Asn Lys His Gly
565 570 575
Ile Glu Glu Glu Pro Phe Leu Asn Glu Phe Val Lys Phe Pro Pro Phe
580 585 590
Lys Ser Asp Tyr Gly Ala Tyr Ser Phe Lys Ala Thr Asn Lys Leu Leu
595 600 605
Ser Leu Met Arg Arg Gly Cys Tyr Trp Asp Glu Glu Asn Ile Asp Cys
610 615 620
Asn Thr Lys Glu Arg Ile Glu Lys Ile Ile Ser Gly Glu Tyr Asp Pro
625 630 635 640
Glu Ile Asn Asp Arg Val Arg Glu Lys Thr Ile Asn Leu Asn Gly Ile
645 650 655
Ser Asp Phe Gln Gly Leu Pro Thr Trp Leu Ala Cys Tyr Val Val Tyr
660 665 670
Gly Arg His Ser Glu Ile Lys Asp Ile Thr Lys Trp Glu Lys Pro Ser
675 680 685
Asp Ile Asp Asn Tyr Leu Lys Leu Phe Lys Gln His Ser Leu Arg Asn
690 695 700
Pro Ile Val Glu Gln Val Val Leu Glu Thr Leu Arg Thr Val Arg Asp
705 710 715 720
Ile Trp Lys Gln Val Gly Arg Ile Asp Glu Ile His Ile Glu Leu Gly
725 730 735
Arg Glu Met Lys Asn Ser Ala Ser Glu Arg Lys Arg Ile Ala Glu Gln
740 745 750
Ile Ser Lys Asn Glu Asn Thr Asn Leu Arg Ile Lys Ala Met Leu Thr
755 760 765
Glu Phe Leu Asn Pro Glu Phe Glu Ile Asp Asn Val Arg Pro Tyr Ser
770 775 780
Pro Thr Gln Gln Glu Ile Leu Lys Ile Tyr Glu Glu Gly Val Leu Asn
785 790 795 800
Ser Gly Ile Glu Ile Asp Glu Lys Val Lys Asn Phe Leu Lys Ser Phe
805 810 815
Asp Lys Ala Glu Asn Arg Pro Thr Arg Ala Glu Phe Leu Lys Tyr Lys
820 825 830
Leu Trp Leu Asp Gln Lys Tyr Ile Ser Pro Tyr Thr Gly Gln Pro Ile
835 840 845
Pro Leu Ser Lys Leu Phe Thr Ser Glu Tyr Glu Ile Glu His Ile Ile
850 855 860
Pro Gln Ser Arg Phe Phe Asp Asp Ser Leu Ser Asn Lys Val Ile Cys
865 870 875 880
Glu Ala Lys Val Asn Ser Glu Lys Gly Ala Arg Leu Gly His Glu Phe
885 890 895
Ile Lys Gly Cys His Glu Gln Ile Ile Asp Leu Gly Phe Gly Lys Thr
900 905 910
Val Lys Ile Leu Ser Ile Glu Ala Tyr Glu Glu His Val Arg Lys Asn
915 920 925
Tyr Gly His Asn Lys Ala Lys Gln Lys Lys Leu Met Leu Asp Glu Ile
930 935 940
Pro Asp Thr Phe Ile Glu Arg Gln Leu Asn Asp Ser Arg Tyr Ile Ser
945 950 955 960
Lys Leu Val Lys Thr Leu Leu Ser Asn Ile Val Arg Glu Asp Asp Glu
965 970 975
Ala Glu Ala Ile Ser Lys Asn Val Ile Thr Cys Thr Gly Gln Ile Thr
980 985 990
Asp Arg Leu Lys His Asp Trp Gly Val Asn Asp Val Trp Asn Gly Ile
995 1000 1005
Ile Leu Pro Arg Phe Gln Arg Met Glu Lys Leu Gln Pro Gly Lys
1010 1015 1020
Arg Phe Thr Ala Thr Asn Thr Asn Gly Lys Leu Ile Pro Tyr Met
1025 1030 1035
Pro Leu Glu Tyr Gln Lys Gly Phe Ser Ser Lys Arg Ile Asp His
1040 1045 1050
Arg His His Ala Met Asp Ala Ile Val Ile Ala Cys Ala Asn Arg
1055 1060 1065
Asn Ile Val Asn Tyr Leu Asn Asn Glu Ser Ala Arg Ser Asp Ala
1070 1075 1080
Lys Ile Ser Arg Tyr Asp Leu Arg Asn Leu Leu Cys Asp Lys Lys
1085 1090 1095
Lys Gln Asp Asp Ala Gly Asn His Thr Trp Thr Met Lys Ile Pro
1100 1105 1110
Trp Asn Thr Phe Ile Arg Asp Met Arg Lys Ala Leu Glu Gly Ile
1115 1120 1125
Ile Val Ser Phe Lys Gln Asn Leu Arg Val Ile Asn Lys Ser Ser
1130 1135 1140
Asn His Ile Thr Lys Tyr Val Asp Gly Gln Lys Lys Arg Val Pro
1145 1150 1155
Gln Ser Glu Gly Asp Asn Arg Ser Ile Arg Lys Ser Leu His Lys
1160 1165 1170
Asp Thr Val Phe Gly Leu Val Asn Leu Arg Glu Lys Lys Thr Val
1175 1180 1185
Ser Leu Ser Glu Ala Leu Lys Lys Pro Asp Arg Ile Val Asp Lys
1190 1195 1200
Ala Leu Lys His Arg Ile Lys Glu Phe Lys Thr Ala Gly Lys Thr
1205 1210 1215
Asp Thr Asp Ile Lys Lys Leu Leu Lys Asn Gly Pro Asp Lys Val
1220 1225 1230
Glu Ile Tyr Tyr Phe Ser Glu Glu Lys Ser Ile Gly Lys Asp Lys
1235 1240 1245
Ala Arg His Tyr Tyr Ala Ala Arg Thr Thr Ile Leu Ser Leu Glu
1250 1255 1260
Met Asp Lys Ser Lys Ser Tyr Glu Lys Ala Ile Asn Thr Ile Asn
1265 1270 1275
Asn Ile Thr Asp Ser Gly Ile Arg Lys Ile Leu Thr Asn His Leu
1280 1285 1290
Glu Ala Asn Gly Asn Asp Pro Ser Lys Ala Phe Ser Ala Asp Gly
1295 1300 1305
Ile Asp Glu Met Asn Lys Asn Ile Ile Leu Leu Asn Gly Gly Lys
1310 1315 1320
Asn His Lys Pro Ile Tyr Ser Val Arg Lys Tyr Glu Glu Ala Asn
1325 1330 1335
Lys Phe Ala Val Gly Glu Ile Gly Cys Lys Ser Lys Lys Phe Val
1340 1345 1350
Glu Ala Asp Lys Gly Gly Asn Leu Tyr Phe Ala Val Tyr Lys Lys
1355 1360 1365
Asp Asp Asn Ser Arg Ser Phe Arg Thr Ile Pro Leu Asn Glu Val
1370 1375 1380
Ile Asp Arg Leu Lys Asn Lys Met Ser Pro Val Pro Glu Thr Asp
1385 1390 1395
Glu Met Gly Asn Arg Leu Ile Phe Trp Leu Ser Pro Asn Asp Leu
1400 1405 1410
Val Tyr Leu Pro Thr Ala Asp Glu Val Glu Asn Gly Arg Val Thr
1415 1420 1425
Leu Pro Leu Asp Lys Asp Arg Ile Tyr Lys Met Val Ser Ala Asn
1430 1435 1440
Lys Lys Gln Cys Phe Phe Met Pro Ser Asn Thr Ala Asn Pro Ile
1445 1450 1455
Ile Ser Ile Glu Phe Ser Ser Ser Asn Lys Met Glu Arg Ala Ile
1460 1465 1470
Thr Gly Glu Met Ile Lys Glu Thr Cys Ile Pro Leu Lys Thr Asp
1475 1480 1485
Arg Leu Gly Asn Ile Thr Asp Phe Asp Gly Arg Ile Ser
1490 1495 1500
<210> 7
<211> 1211
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 7
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Asp Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ala Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Val Glu Ile Leu Asn
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Ile Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Asp Asn Asn Thr Asp Lys Asp Glu Lys Glu Met Glu Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Ile Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Leu Thr Asn Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Arg Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Leu Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Phe Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Ser Thr Asn Val Gln Val
610 615 620
Asp His Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Thr Leu Glu Lys Glu Leu Ile Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Thr Asp Thr Leu Lys Pro Asn Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Thr Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Asn Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210
<210> 8
<211> 1490
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 8
Met Lys Lys Ile Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Leu Ile Glu His Asn Phe Asp Lys Lys Glu Gly Arg Ile Asp Asp
20 25 30
Leu Gly Val Arg Ile Ile Pro Met Ser Ala Asp Ile Leu Gly Lys Phe
35 40 45
Asp Ala Gly Gln Ser His Ser Gln Thr Ala Glu Arg Thr Gly Tyr Arg
50 55 60
Gly Val Arg Arg Leu Tyr Gln Arg Asp Asn Leu Arg Arg Glu Arg Leu
65 70 75 80
His Arg Val Leu Asn Ile Leu Asp Phe Leu Pro Glu His Tyr Ala Glu
85 90 95
His Ile Asp Phe Glu Lys Arg Leu Gly Gln Phe Lys Glu Gly Lys Glu
100 105 110
Ile Lys Leu Asn Tyr Lys Ser Asn Lys Asp Ser Lys Phe Glu Phe Ile
115 120 125
Phe Lys Ala Ser Tyr Asn Glu Met Leu Ala Ala Phe Lys Lys Tyr Gln
130 135 140
Pro Gly Leu Phe Tyr Val Lys Ala Asn Gly Thr Glu Thr Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Ser Gln Pro
165 170 175
Leu Thr Lys Gln Glu Leu Ala Trp Ile Ile Leu Asn Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Ile Asp Asp Asp Lys Asn
195 200 205
Lys Gln Phe Val Gln Leu Lys Val Lys Glu Val Ile Asp Ser Gly Glu
210 215 220
Ala Val Lys Gly Lys Lys Leu Phe Asn Val Ile Phe Glu Asn Gly Trp
225 230 235 240
Lys Tyr Asp Lys Gln Val Val Lys Thr Glu Asp Trp Ile Gly Arg Thr
245 250 255
Lys Glu Phe Ile Val Thr Thr Lys Thr Leu Lys Ser Gly Glu Ile Lys
260 265 270
Arg Thr Tyr Lys Ala Val Asp Ser Glu Lys Asp Trp Ala Ala Ile Lys
275 280 285
Ala Lys Thr Glu Gln Asp Ile Glu Arg Ser Asn Lys Thr Val Gly Glu
290 295 300
Phe Ile Tyr Glu Ala Leu Leu Gln Asp Pro Thr Gln Lys Ile Arg Gly
305 310 315 320
Lys Leu Val Lys Thr Ile Glu Arg Lys Phe Tyr Lys Ala Glu Leu Arg
325 330 335
Glu Ile Leu Arg Lys Gln Ile Glu Leu Gln Pro Gln Leu Phe Thr Thr
340 345 350
Lys Leu Tyr Asn Ala Cys Ile Lys Glu Leu Tyr Pro Asn Asn Glu Ala
355 360 365
His Arg Asn Ser Ile Lys Asn Arg Asp Phe Leu Tyr Leu Phe Leu Asp
370 375 380
Asp Ile Ile Phe Tyr Gln Arg Pro Leu Lys Ser Gln Lys Ser Asn Ile
385 390 395 400
Ser Gly Cys His Leu Glu Gln Arg Ile Tyr Thr Lys Ile Asn Pro Val
405 410 415
Ser Gly Lys Lys Glu Glu Val Lys Gln Ala Val Lys Ala Ile Pro Lys
420 425 430
Ser His Pro Ile Phe Gln Glu Phe Arg Ile Trp Gln Trp Leu Gln Asn
435 440 445
Leu Lys Ile Tyr Asp Lys Ile Asn Thr Asp Lys Gly Glu Leu Ala Asp
450 455 460
Val Thr Asn Gln Leu Leu Pro Ser Glu Glu Ser Leu Leu Asp Leu Phe
465 470 475 480
Asp Tyr Leu Gln Thr Lys Lys Glu Leu Asp Gln Ser Gly Phe Ile Lys
485 490 495
Tyr Phe Ile Asp Lys Lys Leu Ile Asn Lys Ser Glu Lys Glu Asn Tyr
500 505 510
Arg Trp Asn Tyr Val Glu Asp Lys Lys Tyr Pro Phe Ala Glu Thr Arg
515 520 525
Ala Gln Phe Ile Ser Arg Leu Asn Lys Val Lys Asn Ile Asn Asn Ile
530 535 540
Ser Glu Phe Leu Asn Lys Lys Thr Arg Leu Gly Glu Lys Glu Ser Ser
545 550 555 560
Pro Phe Val Thr Arg Ile Glu Gln Leu Trp His Ile Ile Tyr Ser Val
565 570 575
Ser Asp Ile Asn Glu Tyr Lys Ser Ala Leu Glu Lys Phe Ala Leu Lys
580 585 590
His Asp Ile Asp Lys Glu Ser Phe Val Ala Asn Phe Ile Lys Phe Pro
595 600 605
Pro Phe Lys Ser Asp Tyr Gly Ser Tyr Ser Lys Lys Ala Leu Ser Lys
610 615 620
Leu Leu Pro Leu Met Arg Arg Gly Lys Tyr Trp Asn Glu Ser Asp Ile
625 630 635 640
Ser Asn Lys Val Lys Gln Arg Val Ser Asp Ile Met Glu Arg Val Asn
645 650 655
Ala Leu Asn Leu Lys Glu Asn Tyr Asn Ala Lys Glu Leu Ala Glu Ala
660 665 670
Leu Lys Thr Val Ser Asp Asp Asp Val Lys Lys Gln Leu Ile Lys Ser
675 680 685
Phe Val Pro Phe Lys Asp Lys Asn Pro Leu Lys Gly Leu Asn Thr Tyr
690 695 700
Gln Ala Thr Tyr Leu Val Tyr Gly Arg His Ser Glu Val Gly Asp Ile
705 710 715 720
Gln Ser Trp Lys Thr Pro Glu Asp Ile Asp Thr Tyr Leu Lys Asn Phe
725 730 735
Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr Glu
740 745 750
Thr Leu Arg Val Val Arg Asp Ile Trp Ile His Tyr Gly Lys Ser Gln
755 760 765
Leu Asn Phe Phe Asn Glu Ile His Val Glu Leu Gly Arg Glu Met Lys
770 775 780
Asn Pro Ala Asp Lys Arg Lys Gln Ile Ser Asn Arg Asn Ile Glu Asn
785 790 795 800
Glu Asn Thr Asn Asn Arg Ile Arg Glu Ile Leu Lys Asp Leu Lys Asn
805 810 815
Asp Thr Ser Ile Glu Gly Asp Ile Arg Asp Tyr Ser Pro Ser Gln Gln
820 825 830
Asp Leu Leu Lys Ile Tyr Glu Glu Gly Val Tyr Gln Asn Pro Lys Val
835 840 845
Asp Tyr Ser Lys Val Ser Glu Asp Glu Ile Thr Lys Ile Arg Arg Ser
850 855 860
Asn Ser Pro Thr Pro Lys Glu Ile Gln Arg Tyr Arg Leu Trp Leu Glu
865 870 875 880
Gln Gly Tyr Ile Ser Pro Tyr Thr Gly Lys Pro Ile Pro Leu Ser Lys
885 890 895
Leu Phe Thr His Glu Tyr Gln Ile Glu His Ile Ile Pro Gln Ser Arg
900 905 910
Tyr Phe Asp Asn Ser Leu Ser Asn Lys Ile Ile Cys Glu Ser Ala Val
915 920 925
Asn Glu Asp Lys Asp Asn Lys Thr Ala Tyr Glu Tyr Leu Lys Asn Lys
930 935 940
Ser Gly Asn Val Ile Asn Gly His Lys Leu Leu Arg Ile Glu Glu Tyr
945 950 955 960
Glu Ala His Val Asn Arg Tyr Phe Lys Asn Asn Arg Gln Lys Leu Lys
965 970 975
Asn Leu Leu Ser Glu Asp Ile Pro Glu Gly Phe Ile Asn Arg Gln Leu
980 985 990
Asn Asp Ser Arg Tyr Ile Ser Lys Leu Ile Lys Gly Leu Leu Ser Asn
995 1000 1005
Ile Val Arg Gln Glu Asn Glu Gln Glu Ala Thr Ser Lys Asn Leu
1010 1015 1020
Ile Pro Val Thr Gly Ala Val Thr Ser Lys Leu Lys Asn Asp Trp
1025 1030 1035
Gly Leu Asn Asp Lys Trp Asn Glu Leu Ile Leu Pro Arg Phe Glu
1040 1045 1050
Arg Leu Asn Gln Leu Thr Gln Thr Lys Asn Phe Thr Thr Ser Asn
1055 1060 1065
Thr Asn Gly Asn Thr Ile Pro Thr Val Pro Asp Asp Leu Leu Lys
1070 1075 1080
Gly Phe Ser Lys Lys Arg Ile Asp His Arg His His Ala Leu Asp
1085 1090 1095
Ala Leu Val Val Ala Cys Cys Thr Arg Asn His Val Gln Tyr Leu
1100 1105 1110
Asn Ala Leu Asn Ala Glu Lys Ala Asn Tyr Gly Leu Arg Lys Lys
1115 1120 1125
Leu Leu Ile Val Asn Glu Gln Gly Asp Phe Thr Lys Ile Phe Gln
1130 1135 1140
Met Pro Trp Lys Gly Phe Thr Ser Glu Ala Lys Asn Gln Leu Glu
1145 1150 1155
Lys Thr Val Ile Ser Phe Lys Gln Asn Leu Arg Val Ile Asn Lys
1160 1165 1170
Ala Asn Asn Lys Phe Trp Ser Phe Lys Asp Glu Asn Gly Asn Ile
1175 1180 1185
Asn Leu Asp Lys Asn Gly Arg Pro Val Lys Lys Leu Arg Lys Gln
1190 1195 1200
Thr Lys Gly Asp Asn Trp Ala Ile Arg Lys Ala Met His Lys Glu
1205 1210 1215
Thr Val Ser Gly Lys Ser Asn Ile Glu Thr Pro Lys Gly Lys Ile
1220 1225 1230
Ala Thr Ala Val Arg Gly Ser Leu Ala Asp Ile Lys Asn Glu Lys
1235 1240 1245
His Leu Gly Lys Ile Thr Asp Val Gln Ile Arg Glu Val Ile Leu
1250 1255 1260
Pro Asn His Leu Lys Asn Tyr Val Asp Glu Lys Gly Lys Val Lys
1265 1270 1275
Phe Asp Leu Ala Phe Asn Asp Glu Gly Ile Glu Asp Leu Asn Lys
1280 1285 1290
Asn Ile Ile Ala Leu Asn Asn Gly Lys Lys His Gln Pro Ile Arg
1295 1300 1305
Lys Val Lys Phe Phe Glu Val Gly Ser Lys Phe Ser Ile Ser Glu
1310 1315 1320
Asn Glu Asn Ser Ala Lys Ser Lys Lys Tyr Val Glu Ala Ala Lys
1325 1330 1335
Gly Thr Asn Leu Phe Phe Ala Val Tyr Trp Asp Glu Lys Lys Gln
1340 1345 1350
Lys Arg Asn Tyr Glu Thr Val Pro Leu Asn Glu Val Ile Ala His
1355 1360 1365
Gln Lys Gln Val Ala His Leu Thr Asn Asn Glu Arg Leu Pro Ile
1370 1375 1380
Gln Thr Asn Arg Lys Lys Gly Asp Phe Leu Phe Thr Leu Ser Pro
1385 1390 1395
Asn Asp Leu Val Tyr Val Pro Thr Asp Ala Glu Val Ala Asn Lys
1400 1405 1410
Gln Pro Ile Asp Phe Lys Asn Leu His Gln Asn Gln Val Asn Arg
1415 1420 1425
Ile Tyr Lys Met Val Ser Ser Ser Gly Asn Gln Cys Phe Phe Ile
1430 1435 1440
Lys Asp Lys Ile Ala Thr Ser Ile Trp Asn Lys Asn Glu Phe Ser
1445 1450 1455
Ser Leu Asn Lys Met Glu Lys Asp Ile Asp Gly Asn Met Ile Lys
1460 1465 1470
Glu Arg Cys Ile Lys Leu Asn Val Asp Arg Leu Gly Asn Ile Thr
1475 1480 1485
Lys Ala
1490
<210> 9
<211> 1511
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 9
Met Lys Lys Ile Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Val Val Asn Ala Asp Ala Ile Thr Arg Asn Asp Gly Ser Arg Tyr
20 25 30
Leu Lys Pro Asn Ser Ile Ser Ala Ala Gly Ser Arg Ile Ile Pro Met
35 40 45
Ser Ala Asp Val Leu Gly Asn Phe Glu Ser Gly Ile Thr Val Ser Gln
50 55 60
Thr Lys Asp Arg Thr Asp Lys Arg Met Ala Arg Arg Leu His Glu Arg
65 70 75 80
Ala Leu Leu Arg Arg Glu Arg Leu Leu Arg Ile Leu Ser Leu Met Asp
85 90 95
Phe Leu Pro Lys His Phe Ala Ser Lys Ile Asn Arg Tyr Gly Lys Phe
100 105 110
Thr Asp Asp Ser Glu Pro Lys Leu Ala Trp Arg Lys Asn Thr Glu Gly
115 120 125
Lys Tyr Glu Phe Ile Phe Gln Asp Ala Phe Asn Glu Met Leu Ala Glu
130 135 140
Phe Lys Asp Lys Gln Pro Glu Ile Val Lys Glu Gly Lys Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Glu Lys Ala
165 170 175
Leu Ser Lys Glu Glu Leu Ser Trp Leu Leu Leu Gln Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Glu Asp Ile Pro Gln Asp
195 200 205
Lys Lys Ile Glu Tyr Leu Ala Gln Lys Val Val Lys Val Glu Ala Thr
210 215 220
Asp Gln Lys Lys Gly Asp Asp Ile Trp Tyr Asn Val Tyr Leu Glu Asn
225 230 235 240
Gly Met Ile Tyr Arg Arg Thr Ser Lys Ala Pro Leu Asp Trp Glu Gly
245 250 255
Lys Ile Lys Glu Phe Ile Val Thr Thr Asp Leu Glu Lys Asp Gly Thr
260 265 270
Pro Lys Lys Asp Lys Glu Gly Asn Ile Lys Arg Ser Phe Arg Ala Pro
275 280 285
Gln Glu Asp Asp Trp Thr Leu Leu Lys Lys Lys Thr Glu Ala Asp Ile
290 295 300
Glu Lys Ser Thr Lys Thr Val Gly Cys Tyr Ile Tyr Asp Ser Leu Leu
305 310 315 320
Asn Asn Pro Lys Gln Lys Ile Ile Gly Lys Leu Val Arg Thr Val Glu
325 330 335
Arg Lys Phe Tyr Lys Glu Glu Leu Thr Gln Ile Leu Lys Lys Gln Val
340 345 350
Glu Leu Ile Pro Glu Leu Arg Asn Asp Asn Leu Tyr Lys Gln Cys Ile
355 360 365
Glu Glu Leu Tyr Pro Ile Asn Glu Ala His Arg Asn Thr Ile Ala Lys
370 375 380
Thr Asp Phe Ala Asn Leu Phe Ile Asn Asp Ile Leu Phe Tyr Gln Arg
385 390 395 400
Pro Leu Lys Ser Lys Lys Ser Gln Ile Asp Asn Cys Pro Tyr Glu Glu
405 410 415
His Ile Phe Ile Asp Ser Lys Thr Gly Glu Lys Lys Lys Val Pro Val
420 425 430
Lys Cys Ile Thr Lys Ser Asn Pro Leu Phe Gln Glu Phe Arg Leu Trp
435 440 445
Gln Phe Ile Gln Asn Leu Arg Ile Tyr Gln Arg Glu Lys Glu Ile Asp
450 455 460
Gly Lys Leu Ser Thr Asp Val Asp Ile Thr Ser Glu Cys Leu Lys Ser
465 470 475 480
Glu Glu Asp Tyr Val Arg Leu Phe Asp Trp Leu Asn Asp Arg Glu Ser
485 490 495
Ile Glu Gln Glu Glu Leu Leu Lys Tyr Leu Phe Asn Thr Lys Lys Ser
500 505 510
Lys Asn Lys Glu Asn Pro Tyr Arg Trp Asn Tyr Val Glu Asp Lys Val
515 520 525
Tyr Pro Cys Asn Glu Thr Arg Ala Thr Ile Leu Lys Gly Leu Ser Lys
530 535 540
Cys Gly Ile Asn Ala Ser Val Leu Ser Ser Glu Met Glu Met Ala Leu
545 550 555 560
Trp His Ile Leu Tyr Ser Val Glu Asp Lys Lys Glu Ile Glu Thr Ala
565 570 575
Leu Thr His Phe Ala Gln Lys Gln Gly Trp Asn Gly Glu Phe Ala Ile
580 585 590
Val Phe Ser Lys Leu Lys Pro Phe Lys Lys Asp Tyr Gly Ser Tyr Ser
595 600 605
Glu Lys Ala Ile Lys Lys Leu Leu Ser Leu Met Arg Met Gly Lys Tyr
610 615 620
Trp Asn Gln Asp Asn Ile Asp Lys Asn Thr Leu Asp Arg Ile Asp Lys
625 630 635 640
Ile Ile Asn Gly Glu Tyr Asp Glu Lys Ile Ser Asn Arg Val Arg Asp
645 650 655
Asn Ala Ile Asn Leu Lys Asp Ile Ser Asp Phe Arg Gly Leu Pro Val
660 665 670
Trp Leu Ala Cys Tyr Ile Val Tyr Asp Arg His Ser Glu Ala Lys Asp
675 680 685
Cys Thr Lys Trp Asn Thr Pro Glu Glu Ile Asp Ser Tyr Leu Lys Lys
690 695 700
Phe Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr
705 710 715 720
Glu Thr Leu Arg Thr Val Arg Asp Ile Trp Lys Gln Glu Gly Gln Ile
725 730 735
Asp Glu Ile His Leu Glu Leu Gly Arg Asp Leu Lys Asn Pro Ala Asp
740 745 750
Lys Arg Lys Lys Met Ser Glu Asn Ile Leu Lys Asn Glu Asn Thr Asn
755 760 765
Leu Arg Ile Lys Ala Met Leu Met Glu Phe Met Asn Pro Gly Met Gly
770 775 780
Ile Glu Asn Val Arg Pro Tyr Ser Pro Ser Gln Gln Asp Ile Leu Arg
785 790 795 800
Ile Tyr Glu Glu Asn Ala Leu Glu Asn Leu Thr Lys Asp Asp Glu Glu
805 810 815
Phe Asp Phe Ile Ser Lys Ile Ser Lys Gln Ala Gln Pro Thr Lys Ser
820 825 830
Asp Ile Val Arg Tyr Lys Cys Trp Leu Glu Gln Lys Tyr Arg Ser Pro
835 840 845
Tyr Thr Gly Lys Thr Ile Ser Leu Ser Lys Leu Phe Thr Ser Ala Tyr
850 855 860
Glu Ile Glu His Ile Ile Pro Gln Ser Arg Tyr Phe Asp Asp Ser Phe
865 870 875 880
Ser Asn Lys Val Ile Cys Glu Ala Glu Val Asn Lys Leu Lys Asp Arg
885 890 895
Gln Leu Gly His Glu Phe Ile Glu Glu His His Gly Glu Lys Val Gln
900 905 910
Leu Ser Gln Gly Glu Val Val Glu Ile Leu Ser Val Asp Ala Tyr Glu
915 920 925
Lys Phe Val Lys Glu Asn Tyr Ala Asn Asn Arg Val Lys Met Lys Lys
930 935 940
Leu Leu Met Glu Asn Ile Pro Asp Glu Phe Ile Glu Arg Gln Leu Asn
945 950 955 960
Asp Ser Arg Tyr Ile Ser Lys Val Val Lys Gly Leu Leu Ser Asn Ile
965 970 975
Val Arg Glu Lys Ile Asp Asp Glu Asn Tyr Glu Pro Glu Ala Val Ser
980 985 990
Lys Asn Leu Ile Ser Cys Asn Gly Ala Val Thr Asp Arg Leu Lys Lys
995 1000 1005
Asp Trp Gly Met Asn Asp Val Trp Asn Ser Ile Ile Leu Pro Arg
1010 1015 1020
Phe Ile Arg Met Asn Gln Ile Thr Gly Lys Asp Cys Phe Thr Thr
1025 1030 1035
Thr Asn Ala Glu Gly His Leu Ile Pro Gln Met Pro Leu Glu Leu
1040 1045 1050
Gln Lys Gly Phe Asn Lys Lys Arg Ile Asp His Arg His His Ala
1055 1060 1065
Met Asp Ala Ile Val Ile Ala Cys Thr Thr Arg Asp His Val Asn
1070 1075 1080
Leu Leu Asn Asn Glu Ala Ala His Ser Lys Phe Asn Ala Thr Arg
1085 1090 1095
Tyr Gln Leu Gln Arg Lys Leu Arg Cys Phe Glu Lys Ala Met Ile
1100 1105 1110
Asp Gly Lys Glu Arg Glu Val Ala Lys Glu Phe Leu Lys Pro Trp
1115 1120 1125
Asp Ser Phe Thr Met Asp Ser Lys Asn Ile Leu Glu Asn Ile Ile
1130 1135 1140
Val Ser Phe Lys Gln Asn Gln Arg Val Ile Asn Lys Thr Thr Asn
1145 1150 1155
Thr Phe Gln His Phe Asp Glu Asn Gly Lys Lys Thr Phe Val Lys
1160 1165 1170
Gln Gly Lys Gly Asn Ser Trp Ala Ile Arg Lys Pro Met His Lys
1175 1180 1185
Asp Thr Val Phe Gly Glu Ile Asn Leu Arg Lys Val Lys Ser Val
1190 1195 1200
Ser Leu Ser Asp Ala Ile Lys Val Pro Glu Arg Ile Leu Asn Lys
1205 1210 1215
Arg Ile Lys Glu Lys Ile Thr Glu Leu Lys Asn Asn Lys Val Asp
1220 1225 1230
Ala Lys Asn Ile Lys Lys Tyr Ile Glu Glu Tyr His Ile Gly Gly
1235 1240 1245
Tyr Gly Ile Asp Thr Ser Lys Ile Asp Val Phe Tyr Phe Thr Lys
1250 1255 1260
Glu Thr Lys Glu Arg Phe Phe Ala Thr Arg Lys Ser Leu Asp Ser
1265 1270 1275
Ser Phe Asn Gln Ala Lys Ile Glu Asp Ser Ile Ala Asp Ser Gly
1280 1285 1290
Ile Gln Lys Ile Leu Leu Ala His Leu Lys Ser Lys Asn Gly Asp
1295 1300 1305
Ala Glu Gln Ala Phe Ser Pro Asp Gly Ile Asp Glu Met Asn Lys
1310 1315 1320
Asn Ile Val Glu Leu Asn Asn Gly Lys Phe His Gln Pro Ile Leu
1325 1330 1335
Lys Val Arg Val Tyr Glu Lys Ala Asp Lys Phe Ala Val Gly Gln
1340 1345 1350
Lys Gly Asn Lys Lys Val Lys Phe Val Glu Ala Ala Lys Gly Thr
1355 1360 1365
Asn Leu Phe Phe Ala Val Phe Glu Lys Asp Gly Lys Arg Ser Tyr
1370 1375 1380
Leu Thr Ile Pro Leu Asn Val Met Ile Asp Cys Gln Lys Lys Tyr
1385 1390 1395
Gly Asn Gln Trp Lys Gln Asn Ile Glu Ser Tyr Leu Lys Glu Lys
1400 1405 1410
Asp Leu Val Glu Lys Asp Val Gln Leu Leu Phe Ile Leu Ser Pro
1415 1420 1425
Asn Asp Leu Val Tyr Leu Pro Thr Glu Asn Glu Leu Lys Lys Gly
1430 1435 1440
Ile Thr Asn Pro Asp Lys Asp Gln Ile Tyr Lys Phe Val Ser Cys
1445 1450 1455
Thr Ser Asn Glu Ala His Phe Ile Pro Ser Phe Val Ala Asn Pro
1460 1465 1470
Ile Val Gln Thr Thr Glu Leu Gly Ser Asn Asn Lys Ala Gln Arg
1475 1480 1485
Ala Trp Asn Asn Lys Met Ile Lys Glu Ile Cys Ile Pro Ile Glu
1490 1495 1500
Val Asp Arg Leu Gly Asn Ile Lys
1505 1510
<210> 10
<211> 1048
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 10
Met Val Glu Lys Ile Leu Gly Ile Asp Leu Gly Ile Ser Ser Leu Gly
1 5 10 15
Trp Ala Val Val Glu Tyr Asp Lys Asp Asn Asp Glu Asn Asn Lys Ile
20 25 30
Ile Asp Cys Gly Val Arg Leu Phe Thr Ala Ala Glu Thr Pro Lys Glu
35 40 45
Lys Glu Ser Pro Asn Lys Ala Arg Arg Asp Ala Arg Gly Leu Arg Arg
50 55 60
Val Ile Lys Arg Arg Arg Val Arg Met Asn Thr Ile Lys Asn Leu Leu
65 70 75 80
Ile Thr Tyr Lys Leu Ile Asp Lys Thr Leu Leu Asp Glu Glu Met Gly
85 90 95
Met Phe His Ser Gln Ser Asn Arg Val Asp Val Trp Lys Leu Arg His
100 105 110
Asp Ala Leu Tyr His Leu Leu Ser Gly Asp Glu Leu Ala Arg Val Leu
115 120 125
Ile His Ile Ala Lys His Arg Gly Tyr Lys Phe Leu Gly Asp Asp Glu
130 135 140
Ser Asp Glu Glu Ser Gly Lys Val Lys Lys Ala Gly Ala Glu Leu Lys
145 150 155 160
Lys Lys Phe Leu Glu Ala Gly Cys Gln Ser Val Gly Glu Trp Leu Trp
165 170 175
Lys Glu Arg Gly Leu Gln Gly Lys Lys Arg Asn Lys Ser Gly Asp Tyr
180 185 190
Glu Ile Ser Ile Pro Arg Asp Phe Leu Val Glu Glu Ile Gln Arg Ile
195 200 205
Phe Glu Thr Gln Gln Lys Phe Gly Ser Thr Phe Ala Thr Ser Glu Leu
210 215 220
Gln Lys Ala Tyr Thr Asp Ile Ala Phe Tyr Val Lys Pro Met Gln Ser
225 230 235 240
Ile Glu Asp Met Val Gly Tyr Cys Thr Phe Tyr Pro Lys Ile Lys Ser
245 250 255
Lys Asn Gln Asp Gly Glu Lys Arg Ala Pro Lys Ala Ser Pro Ser Ala
260 265 270
Glu Gln Phe Val Ile Leu Ser Lys Ile Phe Ser Thr Ile Val Ile Asp
275 280 285
Glu Asn Lys Gln Glu Lys Lys Leu Ile Glu Leu Lys Ser Ile Glu Gln
290 295 300
Leu Ile Gln Ile Ala Arg Ser Lys Glu Thr Leu Lys Tyr Lys Gln Leu
305 310 315 320
Arg Lys Glu Leu Asn Leu Ala Lys Asp Ile Ser Phe Lys Ser Ile Ser
325 330 335
Asp Glu Glu Lys Thr Trp Ile Asn Leu Val Gly Asn Ala Lys Phe Lys
340 345 350
Lys Ile Leu Gly Leu Asn Tyr Glu Thr Phe Leu Lys Asn Thr Glu Ile
355 360 365
Ser Asp Glu Ile Ala Lys Ile Leu Thr Tyr Asp Lys Thr Phe Glu Gln
370 375 380
Lys Glu Thr Lys Leu Lys Asn Leu Leu Val Asn Ile Asp Trp Ile Asp
385 390 395 400
Asn Asn His Ile Ala Glu Leu Ala Lys Leu Ser Phe Ser Gln Phe Asn
405 410 415
Gln Leu Ser Leu Lys Ala Ile Lys Ile Ile Ser Lys Ile Met Ile Glu
420 425 430
Gly Tyr Ala Arg Tyr Asp Glu Ala Val Gln Tyr Ala Phe Glu Asn Asn
435 440 445
Leu Leu Pro Lys Pro Ser His Glu Lys Ser Ile Leu Leu Pro Pro Leu
450 455 460
Lys Glu Thr Asn Ile Ala Ile Leu Asn Pro Thr Val Ile Arg Ala Phe
465 470 475 480
Ala Gln Phe Arg Gln Val Ala Asn Ala Leu Val Ser Lys Tyr Gly Ser
485 490 495
Phe Asp Lys Val His Phe Glu Leu Ala Arg Glu Val Asn Thr Lys Glu
500 505 510
Asp Arg Lys Arg Trp Glu Lys Asp Arg Asp Lys Asn Glu Lys Met His
515 520 525
Arg Gln Ile Thr Glu Lys Leu Val Glu Glu Gly Val Lys Pro Ser Tyr
530 535 540
Lys Asn Ile Leu Lys Ser Lys Leu Arg Ser Glu Gln Lys Asp Thr Cys
545 550 555 560
Pro Tyr Cys Gln Lys Asn Leu His Tyr Pro Met Ile Phe Glu Asp Gly
565 570 575
Tyr Ala Glu Ile Asp His Ile Leu Pro Leu Ser Gln Ser Gln Asp Asp
580 585 590
Ser Tyr Val Asn Lys Val Leu Val His Ser Ala Cys Asn Gln Asn Lys
595 600 605
Lys Asn Arg Thr Pro Phe Glu Trp Phe Gln Asp Glu Lys Lys Asp Trp
610 615 620
Asp Thr Phe Lys Ser Tyr Ile Leu Met Glu Ser Thr Leu Gly Glu Lys
625 630 635 640
Lys Arg Asn Tyr Leu Ile Lys Glu Asn Phe Ser Asp Pro Gln Ser Arg
645 650 655
Lys Glu Phe Ile Ser Arg Asn Leu Asn Asp Thr Arg Tyr Met Ser Lys
660 665 670
Ala Ile Lys Thr Tyr Cys Glu Asn His Trp Lys Leu Ser His Asp Asp
675 680 685
Asp Lys Leu Arg Ile Gln Val Arg Ser Gly Lys Leu Thr Ser Thr Leu
690 695 700
Arg His Gln Trp Gly Leu Asp Asn Lys Asn Arg Glu Thr His Thr His
705 710 715 720
His Ala Met Asp Ala Ile Met Ile Ala Phe Ser Thr Gln Gly Met Val
725 730 735
Lys Lys Leu Ser Asp Tyr Phe Ala Lys Lys Glu Ala Lys Val Glu Lys
740 745 750
Asp Lys Pro Val Leu Ile Thr Pro Ile Lys Gln Phe Lys Glu Ala Val
755 760 765
Glu Gln Ala Thr Thr Leu Glu Arg Gln Glu Ser Ile Gln Thr Lys Ala
770 775 780
Gly Asp Thr Ile Thr Leu Asn Arg Leu Leu Ile Ser Arg Pro Pro Arg
785 790 795 800
Ala Ser Val Thr Gly Ala Ala His Glu Gln Thr Ala Lys Pro Tyr Pro
805 810 815
Arg Ile Lys Pro Ile Lys Asn Lys Tyr Lys Arg Arg Arg Ile Pro Ile
820 825 830
Asp Glu Asp Lys Phe Glu Leu Phe Arg Asn Asp Lys Val Ala Ser Gly
835 840 845
Asn Asp Lys Asn Phe Tyr Asn Ser Ser Thr Ile Pro Arg Val Asp Ile
850 855 860
Tyr Lys Lys Asp Asp Lys Tyr His Val Val Pro Ile Tyr Leu Ser Asp
865 870 875 880
Met Thr Lys Ala Glu Val Pro Asn Lys Ser Leu Gly Thr Asn Pro Glu
885 890 895
Gly Met Asp Glu Lys Tyr Phe Cys Phe Ser Val Phe Lys Asn Asp Leu
900 905 910
Ile Glu Leu Glu Thr Lys Ala Thr Pro Lys Lys Pro Ser Lys Lys Leu
915 920 925
Leu Gly Tyr Phe Lys Gln Leu Asn Gly Ala Asn Phe Ile Leu Asn Ser
930 935 940
Ile His Asn Gly Ile Ile Asp Gly Phe Val Cys Ser Pro Ile Thr Leu
945 950 955 960
Phe Lys Gln Gln Lys Asp Met Cys Lys Lys Cys Leu Pro Glu Asp Arg
965 970 975
Ala Ile Gly Asn Cys Ser Gln Glu Thr Leu Glu Phe Trp Glu Ala Glu
980 985 990
Asn Ile Lys Val Pro Lys Lys Asp Phe Glu Cys Asp Gln Gly Ile Lys
995 1000 1005
Phe Ala Ile Ala Val Arg Lys Tyr Thr Ile Asp Pro Leu Gly Tyr
1010 1015 1020
Tyr His Glu Val Lys Gly Glu Lys Leu Leu Gly Thr Ile Pro Gln
1025 1030 1035
Gly Ala Lys Lys His Pro Lys Arg Gln Lys
1040 1045
<210> 11
<211> 1337
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 11
Met Lys Lys Asp Tyr Val Ile Gly Leu Asp Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Met Thr Glu Asp Tyr Gln Leu Val Lys Lys Lys Met
20 25 30
Pro Ile Tyr Gly Asn Thr Glu Lys Lys Lys Ile Lys Lys Asn Phe Trp
35 40 45
Gly Val Arg Leu Phe Glu Glu Gly His Thr Ala Glu Asp Arg Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Ile Ser Arg Arg Arg Asn Arg Leu Arg
65 70 75 80
Tyr Leu Gln Ala Phe Phe Glu Glu Ala Met Thr Ala Leu Asp Glu Asn
85 90 95
Phe Phe Ala Arg Leu Gln Glu Ser Phe Leu Val Pro Glu Asp Lys Lys
100 105 110
Trp His Arg His Pro Ile Phe Ala Lys Leu Glu Asp Glu Val Ala Tyr
115 120 125
His Glu Thr Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp
130 135 140
Ser Ser Glu Gln Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His
145 150 155 160
Ile Val Lys Tyr Arg Gly His Phe Leu Ile Glu Gly Lys Leu Ser Thr
165 170 175
Glu Asn Ile Ser Val Lys Glu Gln Phe Gln Gln Phe Met Ile Ile Tyr
180 185 190
Asn Gln Thr Phe Val Asn Gly Glu Ser Arg Leu Val Ser Ala Pro Leu
195 200 205
Pro Glu Ser Val Leu Ile Glu Glu Glu Leu Thr Glu Lys Ala Ser Arg
210 215 220
Thr Lys Lys Ser Glu Lys Val Leu Gln Gln Phe Pro Gln Glu Lys Ala
225 230 235 240
Asn Gly Leu Phe Gly Gln Phe Leu Lys Leu Met Val Gly Asn Lys Ala
245 250 255
Asp Phe Lys Lys Val Phe Gly Leu Glu Glu Glu Ala Lys Ile Thr Tyr
260 265 270
Ala Ser Glu Ser Tyr Glu Glu Asp Leu Glu Gly Ile Leu Ala Lys Val
275 280 285
Gly Asp Glu Tyr Ser Asp Val Phe Leu Ala Ala Lys Asn Val Tyr Asp
290 295 300
Ala Val Glu Leu Ser Thr Ile Leu Ala Asp Ser Asp Lys Lys Ser His
305 310 315 320
Ala Lys Leu Ser Ser Ser Met Ile Val Arg Phe Thr Glu His Gln Glu
325 330 335
Asp Leu Lys Asn Phe Lys Arg Phe Ile Arg Glu Asn Cys Pro Asp Glu
340 345 350
Tyr Asp Asn Leu Phe Lys Asn Glu Gln Lys Asp Gly Tyr Ala Gly Tyr
355 360 365
Ile Ala His Ala Gly Lys Val Ser Gln Leu Lys Phe Tyr Gln Tyr Val
370 375 380
Lys Lys Ile Ile Gln Asp Ile Ala Gly Ala Glu Tyr Phe Leu Glu Lys
385 390 395 400
Ile Ala Gln Glu Asn Phe Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly
405 410 415
Val Ile Pro His Gln Ile His Leu Ala Glu Leu Gln Ala Ile Ile His
420 425 430
Arg Gln Ala Ala Tyr Tyr Pro Phe Leu Lys Glu Asn Gln Lys Lys Ile
435 440 445
Glu Gln Leu Val Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ser
450 455 460
Lys Gly Asp Ala Ser Thr Phe Ala Trp Leu Lys Arg Gln Ser Glu Glu
465 470 475 480
Pro Ile Arg Pro Trp Asn Leu Gln Glu Thr Val Asp Leu Asp Gln Ser
485 490 495
Ala Thr Ala Phe Ile Glu Arg Met Thr Asn Phe Asp Thr Tyr Leu Pro
500 505 510
Ser Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Lys Phe Met
515 520 525
Val Phe Asn Glu Leu Thr Lys Ile Ser Tyr Thr Asp Asp Arg Gly Ile
530 535 540
Lys Ala Asn Phe Ser Gly Lys Glu Lys Glu Lys Ile Phe Asp Tyr Leu
545 550 555 560
Phe Lys Thr Arg Arg Lys Val Lys Lys Lys Asp Ile Ile Gln Phe Tyr
565 570 575
Arg Asn Glu Tyr Asn Thr Glu Ile Val Thr Leu Ser Gly Leu Glu Glu
580 585 590
Asp Gln Phe Asn Ala Ser Phe Ser Thr Tyr Gln Asp Leu Leu Lys Cys
595 600 605
Gly Leu Thr Arg Ala Glu Leu Asp His Pro Asp Asn Ala Glu Lys Leu
610 615 620
Glu Asp Ile Ile Lys Ile Leu Thr Ile Phe Glu Asp Arg Gln Arg Ile
625 630 635 640
Arg Thr Gln Leu Ser Thr Phe Lys Gly Gln Phe Ser Glu Glu Val Leu
645 650 655
Lys Lys Leu Glu Arg Lys His Tyr Thr Gly Trp Gly Arg Leu Ser Lys
660 665 670
Lys Leu Ile Asn Gly Ile Tyr Asp Lys Glu Ser Gly Lys Thr Ile Leu
675 680 685
Asp Tyr Leu Ile Lys Asp Asp Gly Val Ser Lys His Tyr Asn Arg Asn
690 695 700
Phe Met Gln Leu Ile Asn Asp Ser Gln Leu Ser Phe Lys Asn Ala Ile
705 710 715 720
Gln Lys Ala Gln Ser Ser Glu His Glu Glu Thr Leu Ser Glu Thr Val
725 730 735
Asn Glu Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Tyr Gln Ser
740 745 750
Leu Lys Ile Val Asp Glu Leu Val Ala Ile Met Gly Tyr Ala Pro Lys
755 760 765
Arg Ile Val Val Glu Met Ala Arg Glu Asn Gln Thr Thr Ser Thr Gly
770 775 780
Lys Arg Arg Ser Ile Gln Arg Leu Lys Ile Val Glu Lys Ala Met Ala
785 790 795 800
Glu Ile Gly Ser Asn Leu Leu Lys Glu Gln Pro Thr Thr Asn Glu Gln
805 810 815
Leu Arg Asp Thr Arg Leu Phe Leu Tyr Tyr Met Gln Asn Gly Lys Asp
820 825 830
Met Tyr Thr Gly Asp Glu Leu Ser Leu His Arg Leu Ser His Tyr Asp
835 840 845
Ile Asp His Ile Ile Pro Gln Ser Phe Met Lys Asp Asp Ser Leu Asp
850 855 860
Asn Leu Val Leu Val Gly Ser Thr Glu Asn Arg Gly Lys Ser Asp Asp
865 870 875 880
Val Pro Ser Lys Glu Val Val Lys Lys Met Lys Ala Tyr Trp Glu Lys
885 890 895
Leu Tyr Ala Ala Gly Leu Ile Ser Gln Arg Lys Phe Gln Arg Leu Thr
900 905 910
Lys Gly Glu Gln Gly Gly Leu Thr Leu Glu Asp Lys Ala His Phe Ile
915 920 925
Gln Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys Asn Val Ala Gly
930 935 940
Ile Leu Asp Gln Arg Tyr Asn Ala Lys Ser Lys Glu Lys Lys Val Gln
945 950 955 960
Ile Ile Thr Leu Lys Ala Ser Leu Thr Ser Gln Phe Arg Ser Ile Phe
965 970 975
Gly Leu Tyr Lys Val Arg Glu Val Asn Asp Tyr His His Gly Gln Asp
980 985 990
Ala Tyr Leu Asn Cys Val Val Ala Thr Thr Leu Leu Lys Val Tyr Pro
995 1000 1005
Asn Leu Ala Pro Glu Phe Val Tyr Gly Glu Tyr Pro Lys Phe Gln
1010 1015 1020
Ala Phe Lys Glu Asn Lys Ala Thr Ala Lys Ala Ile Ile Tyr Thr
1025 1030 1035
Asn Leu Leu Arg Phe Phe Thr Glu Asp Glu Pro Arg Phe Thr Lys
1040 1045 1050
Asp Gly Glu Ile Leu Trp Ser Asn Ser Tyr Leu Lys Thr Ile Lys
1055 1060 1065
Lys Glu Leu Asn Tyr His Gln Met Asn Ile Val Lys Lys Val Glu
1070 1075 1080
Val Gln Lys Gly Gly Phe Ser Lys Glu Ser Ile Lys Pro Lys Gly
1085 1090 1095
Pro Ser Asn Lys Leu Ile Pro Val Lys Asn Gly Leu Asp Pro Gln
1100 1105 1110
Lys Tyr Gly Gly Phe Asp Ser Pro Val Val Ala Tyr Thr Val Leu
1115 1120 1125
Phe Thr His Glu Lys Gly Lys Lys Pro Leu Ile Lys Gln Glu Ile
1130 1135 1140
Leu Gly Ile Thr Ile Met Glu Lys Thr Arg Phe Glu Gln Asn Pro
1145 1150 1155
Ile Leu Phe Leu Glu Glu Lys Gly Phe Leu Arg Pro Arg Val Leu
1160 1165 1170
Met Lys Leu Pro Lys Tyr Thr Leu Tyr Glu Phe Pro Glu Gly Arg
1175 1180 1185
Arg Arg Leu Leu Ala Ser Ala Lys Glu Ala Gln Lys Gly Asn Gln
1190 1195 1200
Met Val Leu Pro Glu His Leu Leu Thr Leu Leu Tyr His Ala Lys
1205 1210 1215
Gln Cys Leu Leu Pro Asn Gln Ser Glu Ser Leu Ala Tyr Val Glu
1220 1225 1230
Gln His Gln Pro Glu Phe Gln Glu Ile Leu Glu Arg Val Val Asp
1235 1240 1245
Phe Ala Glu Val His Thr Leu Ala Lys Ser Lys Val Gln Gln Ile
1250 1255 1260
Val Lys Leu Phe Glu Ala Asn Gln Thr Ala Asp Val Lys Glu Ile
1265 1270 1275
Ala Ala Ser Phe Ile Gln Leu Met Gln Phe Asn Ala Met Gly Ala
1280 1285 1290
Pro Ser Thr Phe Lys Phe Phe Gln Lys Asp Ile Glu Arg Ala Arg
1295 1300 1305
Tyr Thr Ser Ile Lys Glu Ile Phe Asp Ala Thr Ile Ile Tyr Gln
1310 1315 1320
Ser Thr Thr Gly Leu Tyr Glu Thr Arg Arg Lys Val Val Asp
1325 1330 1335
<210> 12
<211> 1087
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 12
Met Ala Leu Asn Pro Pro Leu Pro Tyr Thr Leu Gly Leu Asp Ile Gly
1 5 10 15
Met Ala Ser Val Gly Ala Ala Leu Leu Thr Glu Gln Arg Ile Leu Gly
20 25 30
Leu His Val Arg Ala Phe Asp Lys Ala Glu Thr Ala Lys Glu Gly Asp
35 40 45
Pro Leu Asn Lys Thr Arg Arg Glu Ser Arg Leu Thr Arg Arg Arg Ile
50 55 60
Arg Arg Arg Ala His Arg Leu Leu Arg Leu Ala Arg Leu Phe Lys Arg
65 70 75 80
Thr Gly Leu Ile Ala Ala Ala His Pro Glu Ala Phe Ala Leu Pro Gly
85 90 95
Ile Ser Pro Trp Asp Leu Arg Ala Asp Gly Leu Asn Arg Leu Leu Thr
100 105 110
Pro Ala Glu Trp Ala Ala Val Leu Tyr His Leu Val Lys His Arg Gly
115 120 125
Phe Gln Ser Thr Arg Lys Ser Glu Ala Lys Glu Asp Glu Lys Ala Gly
130 135 140
Gln Met Leu Ser Gly Val Ser Ala Asn Gln Gln Arg Met Lys Glu Lys
145 150 155 160
Gly Trp Arg Thr Val Gly Glu Met Ala Ala Arg Asp Glu Ala Phe Ala
165 170 175
Glu Ala Lys Arg Asn Lys Gly Gly Ala Tyr Thr His Thr Phe Ala Arg
180 185 190
Thr Asp Leu Val Ala Glu Leu Lys Leu Leu Phe Lys Gln Gln Ala Gly
195 200 205
Phe Gly Asn Pro His Val Gly Val Asp Phe Glu Asn Asp Val Glu Gln
210 215 220
Leu Leu Leu Ala Arg Arg Pro Ala Leu Ala Gly Asp Ala Leu Leu Lys
225 230 235 240
Leu Val Gly Lys Cys Pro Phe Glu Pro Thr Glu Tyr Arg Ala Pro Lys
245 250 255
Ala Ser Tyr Ser Ala Glu Arg Phe Val Trp Leu Thr Lys Leu Asn Asn
260 265 270
Leu Arg Ile Ser Glu Val Gly Glu Gln Arg Ala Leu Thr Ala Gly Glu
275 280 285
Arg Gln Ile Leu Leu Asn Gln Pro Tyr Leu Leu Ala Lys Phe Thr Tyr
290 295 300
Lys Gln Ala Arg Gln Arg Leu Ser Leu Ala Asp Thr Ala Lys Phe Thr
305 310 315 320
Val Leu Thr Tyr Arg Gly Asp Lys Asp Pro Glu Ser Thr Thr Phe Phe
325 330 335
Glu Ala Lys Ala Tyr His Glu Leu Arg Lys Ala Tyr Glu Lys Ala Gly
340 345 350
Leu Glu Ser Arg Trp Gln Arg Asp Ala Leu Asp Pro Thr Arg Leu Asp
355 360 365
Arg Leu Ala Trp Ala Leu Thr Cys Tyr Lys Thr Asp Asp Asp Ile Arg
370 375 380
Ala His Leu Ala Glu His Gly Val Glu Pro Glu Ile Thr Glu Ala Val
385 390 395 400
Leu Gly Glu Ser Phe Asp Lys Phe Val Gly Leu Ser Leu Lys Ala Leu
405 410 415
Gly Lys Ile Leu Pro Phe Met Glu Gln Gly Gln Arg Tyr Asp Glu Ala
420 425 430
Val Gln Ser Ala Gly Tyr Ala His His Ser Gln Leu Asn Arg Asp Thr
435 440 445
Thr Lys Asn Gln Tyr Leu Pro Pro Pro Asp Lys Asp Gln Ile Arg Asn
450 455 460
Pro Val Val Tyr Arg Ala Leu Asn Gln Ala Arg Lys Leu Val Asn Ala
465 470 475 480
Ile Val Arg Glu Tyr Gly Ser Pro Ala Ala Ile His Ile Glu Leu Ala
485 490 495
Arg Asp Leu Ser Lys Pro Met Asp Glu Arg Arg Lys Ile Glu Arg Glu
500 505 510
Gln Lys Glu Phe Gln Glu Arg Lys Ala Lys Asp Arg Glu Ala Phe Ile
515 520 525
Glu Gln Phe Ser Phe Asp Pro Lys Gly Leu Asp Leu Gln Lys Tyr Arg
530 535 540
Leu Tyr Arg Glu Gln Met Ser Gln Cys Ala Tyr Ser Gln Lys Ala Ile
545 550 555 560
Asp Val Thr Arg Leu Phe Glu Pro Gly Tyr Ala Glu Ile Asp His Ala
565 570 575
Leu Pro Tyr Ser Arg Ser Tyr Asp Asp Gly Gln Asn Asn Lys Val Leu
580 585 590
Val Leu Thr Ala Glu Asn Arg Asn Lys Gly Asn Arg Thr Pro Tyr Glu
595 600 605
Tyr Leu Asp Gly Ala Ser Asp Ser Pro Gln Trp Gln Arg Phe Glu Ala
610 615 620
Trp Val Leu Gln Asn Lys Ala Tyr Arg Arg Ala Lys Arg Asp Arg Leu
625 630 635 640
Leu Arg Lys His Phe Gly Glu Asp Glu Ala Glu Gly Phe Arg Glu Arg
645 650 655
Asn Leu Ile Asp Thr Arg Tyr Ile Cys Arg Ala Phe Lys Thr Met Val
660 665 670
Glu Asp His Leu Gln Trp His Ala Asp Ser Asp Ala Lys Asn Arg Cys
675 680 685
Val Val Val Ala Gly Gln Leu Thr Ser Leu Leu Arg Ala Arg Trp Gly
690 695 700
Leu Ile Lys Val Arg Glu Asn Gly Asp Leu His His Ala Leu Asp Ala
705 710 715 720
Ala Val Ile Ala Ala Ala Asn Arg Ser Leu Val Lys Arg Met Ala Asp
725 730 735
Tyr Ser Lys Arg Asn Glu Leu Ala Gln Val Arg Asp Arg Tyr Ile Asp
740 745 750
Pro Ala Thr Gly Glu Ile Leu Asp Ile Ala Ala Met Arg Gln Val Glu
755 760 765
Glu His Phe Pro Ser Pro Trp Pro His Phe Arg Ser Glu Leu Leu Ala
770 775 780
Trp Leu Ser Pro Asn Pro Ala His Gly Leu Asp Gly Leu Ala His Tyr
785 790 795 800
Pro Pro Glu Glu Leu Glu His Leu Arg Pro Met Arg Val Ser Arg Ala
805 810 815
Pro Thr Arg Arg Gly Leu Gly Ala Ala His Gln Glu Thr Ile Arg Ser
820 825 830
Val Gly Arg Glu Gly Arg Leu Leu Ala Asp Gly Gln Ser Ala Val Lys
835 840 845
Thr Pro Leu Thr Ala Ile Lys Leu Lys Asp Leu Glu Asn Ile Val Gly
850 855 860
Tyr Ser His Ser His Asn His Ala Met Ile Glu Ala Ile Arg Lys Arg
865 870 875 880
Leu Glu Thr Asn Gly Asn Asp Gly Ala Lys Ala Phe Lys Met Pro Leu
885 890 895
Phe Lys Pro Ser Ala Thr Asn Gly Tyr Asp Ala Asp Lys Ser His Val
900 905 910
Gly Glu Thr Asp Gln Arg Ala Pro Gln Ile Arg Ser Val Lys Leu Leu
915 920 925
Ala Thr Gln Lys Ser Gly Ile Pro Ile Arg Lys Gly Ile Ala Asn Asn
930 935 940
Gly Ser Met Leu Arg Val Asp Val Phe Gly Lys Gly Gly Lys Phe Tyr
945 950 955 960
Ala Val Pro Val Tyr Val Ala Asp Ala Ala Arg Ala Glu Leu Pro Tyr
965 970 975
Arg Ala Val Ala Ala Phe Lys Pro Glu Asn Glu Trp Pro Glu Met Asp
980 985 990
Glu Lys Gln Phe Met Phe Ser Leu His Pro Asn Asp Trp Val Thr Val
995 1000 1005
Lys Leu Lys Ala Glu Thr Ile Ser Gly Tyr Phe Ala Gly Met Asp
1010 1015 1020
Arg Ser Thr Gly Ala Ile Ser Val Trp Ala His Asp Arg Asn Gln
1025 1030 1035
Ser Ile Gly Lys Asp Gly Gln Trp Arg Gly Val Gly Met Lys Thr
1040 1045 1050
Ala Leu Ala Val Glu Lys Tyr His Val Asp Leu Leu Gly Asn Leu
1055 1060 1065
His Arg Val His Thr Glu Met Arg Leu Pro Leu His Gly Ser Lys
1070 1075 1080
Ala Ser Lys Asp
1085
<210> 13
<211> 1123
<212> PRT
<213> 直肠真杆菌(Eubacterium rectale)
<400> 13
Met Ser Lys Ile Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Leu Ile Asp Asp Asn Gln Asn Arg Ile Leu Gly Val Gly Ser Arg
20 25 30
Ile Phe Pro Met Gly Val Glu Asn Leu Gly Asp Gly Asp Gly Glu Val
35 40 45
Ser Lys Asn Ala Ser Arg Thr Gly Ala Arg Gly Val Arg Arg Gln Phe
50 55 60
Phe Arg Arg Arg Leu Arg Lys Lys Val Leu Leu Lys Ala Leu Ser Glu
65 70 75 80
His Asn Met Cys Pro Met Val Thr Ile Asp Phe Glu Asp Trp Lys Lys
85 90 95
Ser Lys Gln Phe Pro Ser Glu Lys Leu Ser Asn Trp Phe Ser Leu Asn
100 105 110
Pro Tyr Glu Leu Arg His Lys Ala Leu Ser Glu Lys Leu Thr Leu Glu
115 120 125
Glu Ile Gly Arg Ile Leu Tyr His Leu Ile Gln Arg Arg Gly Phe Leu
130 135 140
Ser Asn Ser Arg Lys Gly Gly Ser Asp Asp Gly Ala Ile Phe Lys Gly
145 150 155 160
Asn Pro Lys Glu Gly Lys Ile Gly Ile Thr Glu Thr Gln Glu Ser Ile
165 170 175
Gln Asp Lys Ser Leu Gly Ser Tyr Leu Phe Glu Ile Tyr Pro Lys Glu
180 185 190
Asn Gln Pro Phe Glu Gly Gly Leu Glu Arg Ile Arg Asn Arg Tyr Thr
195 200 205
Ile Arg Lys Met Tyr Val Asp Glu Phe Glu Leu Ile Trp Asn Lys Gln
210 215 220
Ser Gln Phe His Ser Ser Leu Asn Asp Asp Leu Lys Thr Leu Leu Gly
225 230 235 240
Gly Arg Lys Leu Asp Gly Tyr Lys Glu Asp Gly Ile Leu Phe His Gln
245 250 255
Arg Pro Leu Arg Ser Gln Lys His Leu Val Gly Asn Cys Ser Phe Glu
260 265 270
Pro Thr Lys Thr Lys Cys Pro Ile Ser Ala Ile Pro Phe Glu Met Phe
275 280 285
Arg Ile Trp Gln Trp Val Asn Thr Leu Glu Tyr Asn Gly Lys Lys Ile
290 295 300
Thr Gln Glu Glu Lys Glu Lys Ile Val Glu Phe Met Cys Ala Asn Glu
305 310 315 320
Lys Pro Asp Phe Lys Arg Ile Arg Lys Val Ile Gly Lys Glu Ser Ala
325 330 335
Glu Phe Lys Phe Asn Tyr Lys Asp Asp Asp Lys Ile Val Gly Thr His
340 345 350
Thr Ile Ser Asn Leu Ser Asn Lys Lys Phe Phe Gly Lys Ala Trp Phe
355 360 365
Asp Phe Ser Glu Lys Glu Gln Glu Asp Ile Trp His Val Leu Tyr Phe
370 375 380
Phe Asp Ser Lys Ser Asn Leu Lys Asp Tyr Ala Ile Lys His Trp Asn
385 390 395 400
Phe Asn Glu Ala Gln Ala Ser Asp Val Ser Lys Phe Asn Val Lys Asp
405 410 415
Gly Tyr Ser Ser Leu Ser Arg Lys Ala Ile Ser Asn Ile Leu Pro Phe
420 425 430
Leu Lys Leu Gly Phe Thr Tyr Asp Val Ser Val Val Leu Gly Gly Ile
435 440 445
Lys Asn Val Phe Gly Ser Glu Trp Glu Lys Leu Ser Glu Glu Lys Arg
450 455 460
Asn Tyr Leu Ile Asp Asn Val Glu Gly Ile Val Arg Ser Lys Ile Lys
465 470 475 480
Gly Gly Phe Ile Asp Val Ile Lys Gly Ile Leu Arg Asn Asp Tyr Ser
485 490 495
Ile Ser Asp Asn Gln Leu Arg Lys Leu Tyr His His Ser Ala Thr Ile
500 505 510
Asp Ala Val Glu Leu Leu Asp Lys Leu Pro Val Gly Lys Glu Ala Asp
515 520 525
Lys Glu Ile Gln Ala Ile Arg Asn Pro Ile Val Ile Thr Ala Leu Phe
530 535 540
Glu Leu Arg Lys Leu Val Asn Glu Leu Ile Asp Glu His Gly Lys Leu
545 550 555 560
Asp Glu Ile Lys Val Glu Met Ala Arg Asp Leu Lys Ile Ser Lys Ser
565 570 575
Gln Arg Asn Lys Ile Arg Arg Glu Gln Lys Arg Leu Glu Arg Glu Asn
580 585 590
Asp Arg Val Lys Asp Arg Leu Val Glu Asn Asn Ile Arg Ile Thr His
595 600 605
Asp Asn Ile Leu Leu Tyr Lys Leu Trp Glu Glu Cys Lys Lys Thr Cys
610 615 620
Pro Tyr Thr Gly Lys Pro Ile Ser Val Thr Gln Leu Phe Ser Gly Glu
625 630 635 640
Val Gln Ile Glu His Ile His Pro Trp Ser Arg Ser Leu Asn Asp Ser
645 650 655
Phe Ser Asn Lys Thr Leu Cys Tyr Ala Asp Glu Asn Arg Lys Lys Gly
660 665 670
Asn Leu Thr Pro Phe Glu Phe Tyr Gly Ser Asp Glu Thr Asn Trp Ser
675 680 685
Ala Ile Lys Glu Arg Ala Leu Lys Leu Phe Ser Asp Thr Lys Glu Tyr
690 695 700
Pro Asn Ala Tyr Gln Lys Phe Lys Arg Phe Val Gln Val Lys Phe Asp
705 710 715 720
Asp Asp Phe Ser Ser Arg Gln Leu Asn Asp Thr Arg Tyr Ile Ser Lys
725 730 735
Glu Ala Lys Asn Tyr Leu Ser Arg Ile Cys Lys Asn Val Ile Val Ser
740 745 750
Pro Gly Gln Ala Thr Ser Asn Leu Arg Gln Lys Trp Gly Met Asn Asn
755 760 765
Ile Leu Ser Asp Glu Asn Glu Lys Thr Arg Asp Asp His Arg His His
770 775 780
Ala Val Asp Ala Leu Val Met Ala Cys Thr Lys Val Ser Tyr Val Gln
785 790 795 800
Glu Leu Ala Lys Trp Asn Arg Tyr Asn Arg Asn Ser Glu Leu Lys Asn
805 810 815
Phe Pro Leu Pro Trp Glu Thr Phe Arg Phe Asp Ala Glu Lys Ala Val
820 825 830
Glu Lys Ile Leu Ile Ser His Lys Lys Val Ser Asn Asp Ile Thr Val
835 840 845
Arg Thr His Ile Thr Glu Lys Asn Gly Ile Lys Tyr Lys Asn Val Gly
850 855 860
Val Ala Ala Arg Gly Gln Leu His Lys Glu Thr Val Phe Gly Lys Arg
865 870 875 880
Thr Phe Asn Gly Glu Glu Ala Tyr His Val Arg Lys Ser Ile Asp Ser
885 890 895
Leu Glu Thr Ala Lys Gln Ile Glu Lys Val Val Asp Glu Thr Ile Lys
900 905 910
Gln Leu Ile Leu Lys Arg Val Asn Glu Leu Gly Gly Phe Val Lys Asp
915 920 925
Lys Val Pro Ala Asn Thr Phe Phe Ile Val Asp Glu Lys Gly Ile Lys
930 935 940
Gln Pro Gln Leu Phe Leu Pro Asn Lys Asn Gly Gln Pro Ile Pro Val
945 950 955 960
Leu Lys Val Arg Val Lys Glu Ser Val Gly Arg Ala Glu Gln Leu Lys
965 970 975
Ala Asn Val Asn Gln Trp Val Asn Pro Arg Asn Asn His His Val Leu
980 985 990
Ile Tyr Lys Asp Glu His Gly Asn Leu Lys Glu Asp Val Val Thr Phe
995 1000 1005
Trp Thr Val Val Glu Arg Lys Arg Thr Gly Gln Ser Ile Tyr Gln
1010 1015 1020
Leu Pro Ile Asn Gly Lys Glu Ile Ile Thr Ser Leu His Thr Asn
1025 1030 1035
Asp Met Phe Ile Ile Gly Leu Asn Glu Asp Glu Ile Asn Trp Glu
1040 1045 1050
Leu Ile Asp Phe Asn Leu Ile Asn His His Leu Tyr Arg Val Gln
1055 1060 1065
Lys Thr Ser Lys Lys Glu Lys Ser Phe Glu Phe Asn Phe Arg Leu
1070 1075 1080
Gly Ile Ala Ser Ser Leu Asp Asn Lys Ser Gln Glu Ile Ser Ile
1085 1090 1095
Gln Ser Phe Lys Lys Trp Ile Glu Leu Asn Pro Ile Lys Val Lys
1100 1105 1110
Ile Ser Val Ser Gly Lys Ile Gln Lys Val
1115 1120
<210> 14
<211> 1322
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 14
Met Lys Asn Thr Asn Glu Asp Tyr Tyr Leu Gly Leu Ala Ile Gly Thr
1 5 10 15
Asp Ser Val Gly Trp Ala Val Thr Asp Lys Glu Tyr Asn Ile Leu Glu
20 25 30
Phe Arg Arg Lys Pro Met Trp Gly Ile His Leu Phe Glu Gly Gly Ser
35 40 45
Thr Ala Gln Lys Thr Arg Val Tyr Arg Thr Ser Arg Arg Arg Leu Lys
50 55 60
Arg Arg Ala Glu Arg Ile Ala Leu Leu Arg Asp Ile Phe Ser Glu Glu
65 70 75 80
Ile Gly Lys Val Asp Pro Gly Phe Phe Glu Arg Leu Asp Glu Ser Asp
85 90 95
Leu His Leu Glu Asp Arg Val Thr Ser Gln Lys Asn Ser Leu Phe Asp
100 105 110
Asp Pro Glu Phe Asn Asp Lys Asp Leu His Lys Arg Phe Pro Thr Ile
115 120 125
Tyr His Leu Arg Arg His Leu Met His Ser Asn Arg Lys Glu Asp Ile
130 135 140
Arg Leu Ile Tyr Leu Ala Ala His His Ile Ile Lys Phe Arg Gly His
145 150 155 160
Phe Leu Tyr Lys Gly Ile Gly Asp Glu Glu Ile Pro Ser Phe Glu Ile
165 170 175
Val Leu Asn Ser Leu Ile Asp Asn Leu Arg Asp Glu Tyr Gly Met Glu
180 185 190
Leu Glu Val Ser Asp Arg Asp Leu Val Lys Ala Leu Leu Ser Asp Phe
195 200 205
Ser Ile Gly Ile Arg Glu Lys Ser Arg Glu Leu Ser Ser Cys Leu Asn
210 215 220
Ala Glu Ser Glu Asn Glu Lys Ala Leu Val Asp Phe Ile Ser Gly Lys
225 230 235 240
Lys Thr Asn Met Lys Lys Leu Phe Asp Asp Glu Ala Leu Asp Lys Met
245 250 255
Ser Phe Ser Leu Arg Asp Ser Gly Phe Glu Asp Gln Leu Arg Glu Asn
260 265 270
Glu Gly Val Leu Gly Pro Glu Arg Val His Thr Leu Glu Leu Ser Arg
275 280 285
Gln Ile Phe Glu Trp Ala Arg Leu Ser Ser Ile Leu Lys Asp Ser Asp
290 295 300
Ser Ile Ser Glu Ala Lys Ile Lys Asp Tyr Asp Gln His Arg Glu Asp
305 310 315 320
Leu Arg Met Leu Lys Arg Ala Val Lys Lys Tyr Ala Pro Asp Lys Tyr
325 330 335
Ser Glu Val Phe Lys Ser Lys Glu His Thr Gly Asn Tyr Cys Ser Tyr
340 345 350
Val Tyr Val Cys Gly Lys Gly Leu Pro Asp Lys Lys Cys Ser Thr Glu
355 360 365
Glu Phe Gln Lys Tyr Leu Lys Lys Ile Leu Asp Asp Ser Gly Val Arg
370 375 380
Asp Asp Glu Glu Phe Lys Thr Leu Ile Gln Arg Leu Asp Ala Gly Ile
385 390 395 400
Leu Cys Pro Lys Gln Arg Thr Gly Glu Asn Ser Val Ile Pro Tyr Ser
405 410 415
Val His Arg Lys Glu Leu Ile Gly Ile Leu Asn Asn Ala Ala Glu His
420 425 430
Tyr Pro Ser Leu Ser Arg Lys Gly Glu Asp Gly Phe Ser Ser Ile Asp
435 440 445
Lys Ile Leu Met Leu Glu Glu Phe Arg Ile Pro Tyr Tyr Val Gly Pro
450 455 460
Leu Asp Asp Arg Ser Ser Arg Ser Trp Leu Ile Arg Asn Ser Phe Glu
465 470 475 480
Ala Ile Thr Pro Trp Asn Phe Asn Glu Ile Val Asp Glu Asp Glu Thr
485 490 495
Ser Glu Arg Phe Ile Gly Asn Leu Thr Ser Met Cys Thr Tyr Leu Gly
500 505 510
Gly Glu Lys Val Leu Pro Lys Asn Ser Leu Leu Tyr Ser Arg Phe Met
515 520 525
Leu Tyr Asn Glu Ile Asn Asn Leu Arg Val Gly Gly Glu Lys Ile Pro
530 535 540
Ala Ala Leu Lys Asn Lys Met Val Ser Glu Leu Phe Ala Asn Arg Ala
545 550 555 560
Thr Ser Ser Lys Val Thr Leu Lys Glu Leu Lys Ala Phe Leu Lys Gly
565 570 575
Glu Gly Val Leu Thr Asp Ala Asp Glu Ile Ser Gly Ile Asp Asp Gly
580 585 590
Val Lys Ser Thr Leu Arg Ser Glu Ile Leu Ile Arg Lys Ile Ile Gly
595 600 605
Asp Lys Ile Ser Asp Arg Glu Met Ala Glu Glu Ile Val Arg Ile Leu
610 615 620
Thr Val Phe Gly Asp Glu Arg Arg Arg Ser Lys Ala Lys Leu Lys Lys
625 630 635 640
Glu Phe Ser Asp Lys Leu Thr Glu Lys Glu Ile Glu Lys Leu Ser Ser
645 650 655
Leu Lys Phe Asp Gly Trp Gly Arg Leu Ser Glu Lys Phe Leu Thr Gly
660 665 670
Leu Arg Gln Glu Val Asn Gly Arg Ser Met Ser Ile Ile Glu Ile Leu
675 680 685
Glu Asp Thr Asn Tyr Asn Leu Gln Glu Thr Leu Ser Lys Tyr Ser Phe
690 695 700
Asn Glu Ile Ile Asp Ser Tyr Asn Glu Val Leu Thr Ser Gly Pro Arg
705 710 715 720
Ser Ile Ser Tyr Asp Ile Leu Lys Asp Ser Tyr Leu Ser Pro Ala Val
725 730 735
Lys Arg Gly Val Trp Arg Ala Leu Ser Val Val Lys Asp Ile Leu Lys
740 745 750
Ala Val Gly Arg Pro Pro Lys Lys Ile Phe Val Glu Thr Thr Arg Glu
755 760 765
Glu Arg Glu Lys Lys Arg Thr Glu Ser Arg Lys Asp Ala Leu Met Tyr
770 775 780
Leu Tyr Lys Ser Cys Lys Glu Thr Glu Trp Glu Lys Arg Leu Asp Ser
785 790 795 800
Val Glu Glu Ser Ser Leu Arg Asn Arg Ser Leu Tyr Leu Tyr Tyr Thr
805 810 815
Gln Leu Gly Lys Cys Met Tyr Cys Gly Lys Asn Ile Asp Ile Gly Glu
820 825 830
Leu Asn Thr Asp Leu Ala Asp Arg Asp His Ile Tyr Pro Gln Ser Lys
835 840 845
Thr Lys Asp Asp Ser Ile Arg Asn Asn Leu Val Leu Val Cys Arg Gly
850 855 860
Cys Asn Gln Ala Lys Gly Asp Arg Tyr Pro Leu Pro Gln Glu Trp Val
865 870 875 880
Ser Arg Met His Ala Phe Trp Thr Met Leu Lys Asp Lys Gly Tyr Ile
885 890 895
Ser Ser Glu Lys Tyr Arg Arg Leu Thr Arg Arg Gly Glu Leu Thr Glu
900 905 910
Glu Glu Phe Gly Ala Phe Ile Asn Arg Gln Leu Val Glu Thr Ser Gln
915 920 925
Ser Ala Lys Ala Val Ile Thr Val Leu Lys Asn Ala Phe Lys Asp Ser
930 935 940
Asp Ile Val Tyr Val Lys Gly Ser Asn Val Ser Asp Phe Arg Ser Ser
945 950 955 960
Tyr Asn Phe Ile Lys Cys Arg Ser Val Asn Asp Tyr His His Ala Lys
965 970 975
Asp Ala Tyr Leu Asn Ile Val Val Gly Asn Val Leu Asp Thr Lys Phe
980 985 990
Thr Lys Asn Pro Ser Tyr Val Leu Lys Asn Arg Glu Gln Tyr Asn Ile
995 1000 1005
Gly Arg Met Tyr Asp Arg Asn Val Ser Arg Phe Gly Val Asp Ala
1010 1015 1020
Trp Val Ala Gly Asp Arg Gly Ser Ile Ala Thr Val Arg Lys Tyr
1025 1030 1035
Met Arg Arg Asn Asn Ile Leu Phe Thr Arg Tyr Ala Thr Lys Ser
1040 1045 1050
Lys Gly Ala Leu Phe Lys Glu Thr Val His Arg Lys Lys Glu Gly
1055 1060 1065
Leu Phe Glu Arg Lys Lys Gly Leu Glu Thr Glu Lys Tyr Gly Gly
1070 1075 1080
Tyr Ser Asp Ile Ser Thr Ser Tyr Leu Thr Leu Leu Glu Tyr Asp
1085 1090 1095
Lys Gly Lys Lys Arg Ile Arg Ser Leu Glu Ile Val Pro Thr Tyr
1100 1105 1110
Phe Ala Asn Thr Arg Pro Lys Glu Glu Asp Val Ile Arg Phe Phe
1115 1120 1125
Ser Glu Thr Arg Gly Leu Ala Asn Val Arg Val Val Met Pro Glu
1130 1135 1140
Val Arg Met Lys Ser Leu Phe Glu Tyr Arg Gly Phe Arg Phe His
1145 1150 1155
Val Thr Gly Ser Asn Gly Lys Gly Arg Phe Trp Ile Ser Ser Ala
1160 1165 1170
Ile Gln Leu Leu Leu Pro Glu Asn Leu Tyr Ala Tyr Cys Lys Ser
1175 1180 1185
Ile Glu Asn Asn Glu Lys Asp Ser Gln Arg Arg Ser Glu Lys Pro
1190 1195 1200
Leu Gln Asn Tyr Gly Phe Ser Ser Glu Met Asn Ile Glu Leu Phe
1205 1210 1215
Lys Cys Leu Met Asp Lys Ala Ala Lys Pro Pro Tyr Asp Val Lys
1220 1225 1230
Leu Ser Thr Leu Ser Lys Asn Leu Glu Glu Gly Phe Glu Lys Phe
1235 1240 1245
Lys Ala Leu Glu Leu Gly Pro Gln Val Lys Val Leu Gln Gln Ile
1250 1255 1260
Leu Asp Ile Tyr Ser Cys Asp Arg Lys Ser Gly Asp Leu Ser Val
1265 1270 1275
Leu Gly Ser Ala Arg Asn Ala Gly Arg Leu Asp Met Asn Gly Val
1280 1285 1290
Leu Ser Glu Ala Asp Gly Glu Gln Val Thr Met Ile Cys Gln Ser
1295 1300 1305
Pro Ser Gly Leu Phe Glu Lys Arg Val Pro Met Asn Glu Lys
1310 1315 1320
<210> 15
<211> 1322
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 15
Met Lys Asn Thr Asn Glu Asp Tyr Tyr Leu Gly Leu Asp Ile Gly Thr
1 5 10 15
Asp Ser Val Gly Trp Ala Val Thr Asp Lys Glu Tyr Asn Ile Leu Glu
20 25 30
Phe Arg Arg Lys Pro Met Trp Gly Ile His Leu Phe Glu Gly Gly Ser
35 40 45
Thr Ala Gln Lys Thr Arg Val Tyr Arg Thr Ser Arg Arg Arg Leu Lys
50 55 60
Arg Arg Ala Glu Arg Ile Ala Leu Leu Arg Asp Ile Phe Ser Glu Glu
65 70 75 80
Ile Gly Lys Val Asp Pro Gly Phe Phe Glu Arg Leu Asp Glu Ser Asp
85 90 95
Leu His Leu Glu Asp Arg Val Thr Ser Gln Lys Asn Ser Leu Phe Asp
100 105 110
Asp Pro Glu Phe Asn Asp Lys Asp Leu His Lys Arg Phe Pro Thr Ile
115 120 125
Tyr His Leu Arg Arg His Leu Met His Ser Asn Arg Lys Glu Asp Ile
130 135 140
Arg Leu Ile Tyr Leu Ala Ala His His Ile Ile Lys Phe Arg Gly His
145 150 155 160
Phe Leu Tyr Lys Gly Ile Gly Asp Glu Glu Ile Pro Ser Phe Glu Ile
165 170 175
Val Leu Asn Ser Leu Ile Asp Asn Leu Arg Asp Glu Tyr Gly Met Glu
180 185 190
Leu Glu Val Ser Asp Arg Asp Leu Val Lys Ala Leu Leu Ser Asp Phe
195 200 205
Ser Ile Gly Ile Arg Glu Lys Ser Arg Glu Leu Ser Ser Cys Leu Asn
210 215 220
Ala Glu Ser Glu Asn Glu Lys Ala Leu Val Asp Phe Ile Ser Gly Lys
225 230 235 240
Lys Thr Asn Met Lys Lys Leu Phe Asp Asp Glu Ala Leu Asp Lys Met
245 250 255
Ser Phe Ser Leu Arg Asp Ser Gly Phe Glu Asp Gln Leu Arg Glu Asn
260 265 270
Glu Gly Val Leu Gly Pro Glu Arg Val His Thr Leu Glu Leu Ser Arg
275 280 285
Gln Ile Phe Glu Trp Ala Arg Leu Ser Ser Ile Leu Lys Asp Ser Asp
290 295 300
Ser Ile Ser Glu Ala Lys Ile Lys Asp Tyr Asp Gln His Arg Glu Asp
305 310 315 320
Leu Arg Met Leu Lys Arg Ala Val Lys Lys Tyr Ala Pro Asp Lys Tyr
325 330 335
Ser Glu Val Phe Lys Ser Lys Glu His Thr Gly Asn Tyr Cys Ser Tyr
340 345 350
Val Tyr Val Cys Gly Lys Gly Leu Pro Asp Lys Lys Cys Ser Thr Glu
355 360 365
Glu Phe Gln Lys Tyr Leu Lys Lys Ile Leu Asp Asp Ser Gly Val Arg
370 375 380
Asp Asp Glu Glu Phe Lys Thr Leu Ile Gln Arg Leu Asp Ala Gly Ile
385 390 395 400
Leu Cys Pro Lys Gln Arg Thr Gly Glu Asn Ser Val Ile Pro Tyr Ser
405 410 415
Val His Arg Lys Glu Leu Ile Gly Ile Leu Asn Asn Ala Ala Glu His
420 425 430
Tyr Pro Ser Leu Ser Arg Lys Gly Glu Asp Gly Phe Ser Ser Ile Asp
435 440 445
Lys Ile Leu Met Leu Glu Glu Phe Arg Ile Pro Tyr Tyr Val Gly Pro
450 455 460
Leu Asp Asp Arg Ser Ser Arg Ser Trp Leu Ile Arg Asn Ser Phe Glu
465 470 475 480
Ala Ile Thr Pro Trp Asn Phe Asn Glu Ile Val Asp Glu Asp Glu Thr
485 490 495
Ser Glu Arg Phe Ile Gly Asn Leu Thr Ser Met Cys Thr Tyr Leu Gly
500 505 510
Gly Glu Lys Val Leu Pro Lys Asn Ser Leu Leu Tyr Ser Arg Phe Met
515 520 525
Leu Tyr Asn Glu Ile Asn Asn Leu Arg Val Gly Gly Glu Lys Ile Pro
530 535 540
Ala Ala Leu Lys Asn Lys Met Val Ser Glu Leu Phe Ala Asn Arg Ala
545 550 555 560
Thr Ser Ser Lys Val Thr Leu Lys Glu Leu Lys Ala Phe Leu Lys Gly
565 570 575
Glu Gly Val Leu Thr Asp Ala Asp Glu Ile Ser Gly Ile Asp Asp Gly
580 585 590
Val Lys Ser Thr Leu Arg Ser Glu Ile Leu Ile Arg Lys Ile Ile Gly
595 600 605
Asp Lys Ile Ser Asp Arg Glu Met Ala Glu Glu Ile Val Arg Ile Leu
610 615 620
Thr Val Phe Gly Asp Glu Arg Arg Arg Ser Lys Ala Lys Leu Lys Lys
625 630 635 640
Glu Phe Ser Asp Lys Leu Thr Glu Lys Glu Ile Glu Lys Leu Ser Ser
645 650 655
Leu Lys Phe Asp Gly Trp Gly Arg Leu Ser Glu Lys Phe Leu Thr Gly
660 665 670
Leu Arg Gln Glu Val Asn Gly Arg Ser Met Ser Ile Ile Glu Ile Leu
675 680 685
Glu Asp Thr Asn Tyr Asn Leu Gln Glu Thr Leu Ser Lys Tyr Ser Phe
690 695 700
Asn Glu Ile Ile Asp Ser Tyr Asn Glu Val Leu Thr Ser Gly Pro Arg
705 710 715 720
Ser Ile Ser Tyr Asp Ile Leu Lys Asp Ser Tyr Leu Ser Pro Ala Val
725 730 735
Lys Arg Gly Val Trp Arg Ala Leu Ser Val Val Lys Asp Ile Leu Lys
740 745 750
Ala Val Gly Arg Pro Pro Lys Lys Ile Phe Val Glu Thr Thr Arg Glu
755 760 765
Glu Arg Glu Lys Lys Arg Thr Glu Ser Arg Lys Asp Ala Leu Met Tyr
770 775 780
Leu Tyr Lys Ser Cys Lys Glu Thr Glu Trp Glu Lys Arg Leu Asp Ser
785 790 795 800
Val Glu Glu Ser Ser Leu Arg Asn Arg Ser Leu Tyr Leu Tyr Tyr Thr
805 810 815
Gln Leu Gly Lys Cys Met Tyr Cys Gly Lys Asn Ile Asp Ile Gly Glu
820 825 830
Leu Asn Thr Asp Leu Ala Asp Arg Asp Ala Ile Tyr Pro Gln Ser Lys
835 840 845
Thr Lys Asp Asp Ser Ile Arg Asn Asn Leu Val Leu Val Cys Arg Gly
850 855 860
Cys Asn Gln Ala Lys Gly Asp Arg Tyr Pro Leu Pro Gln Glu Trp Val
865 870 875 880
Ser Arg Met His Ala Phe Trp Thr Met Leu Lys Asp Lys Gly Tyr Ile
885 890 895
Ser Ser Glu Lys Tyr Arg Arg Leu Thr Arg Arg Gly Glu Leu Thr Glu
900 905 910
Glu Glu Phe Gly Ala Phe Ile Asn Arg Gln Leu Val Glu Thr Ser Gln
915 920 925
Ser Ala Lys Ala Val Ile Thr Val Leu Lys Asn Ala Phe Lys Asp Ser
930 935 940
Asp Ile Val Tyr Val Lys Gly Ser Asn Val Ser Asp Phe Arg Ser Ser
945 950 955 960
Tyr Asn Phe Ile Lys Cys Arg Ser Val Asn Asp Tyr His His Ala Lys
965 970 975
Asp Ala Tyr Leu Asn Ile Val Val Gly Asn Val Leu Asp Thr Lys Phe
980 985 990
Thr Lys Asn Pro Ser Tyr Val Leu Lys Asn Arg Glu Gln Tyr Asn Ile
995 1000 1005
Gly Arg Met Tyr Asp Arg Asn Val Ser Arg Phe Gly Val Asp Ala
1010 1015 1020
Trp Val Ala Gly Asp Arg Gly Ser Ile Ala Thr Val Arg Lys Tyr
1025 1030 1035
Met Arg Arg Asn Asn Ile Leu Phe Thr Arg Tyr Ala Thr Lys Ser
1040 1045 1050
Lys Gly Ala Leu Phe Lys Glu Thr Val His Arg Lys Lys Glu Gly
1055 1060 1065
Leu Phe Glu Arg Lys Lys Gly Leu Glu Thr Glu Lys Tyr Gly Gly
1070 1075 1080
Tyr Ser Asp Ile Ser Thr Ser Tyr Leu Thr Leu Leu Glu Tyr Asp
1085 1090 1095
Lys Gly Lys Lys Arg Ile Arg Ser Leu Glu Ile Val Pro Thr Tyr
1100 1105 1110
Phe Ala Asn Thr Arg Pro Lys Glu Glu Asp Val Ile Arg Phe Phe
1115 1120 1125
Ser Glu Thr Arg Gly Leu Ala Asn Val Arg Val Val Met Pro Glu
1130 1135 1140
Val Arg Met Lys Ser Leu Phe Glu Tyr Arg Gly Phe Arg Phe His
1145 1150 1155
Val Thr Gly Ser Asn Gly Lys Gly Arg Phe Trp Ile Ser Ser Ala
1160 1165 1170
Ile Gln Leu Leu Leu Pro Glu Asn Leu Tyr Ala Tyr Cys Lys Ser
1175 1180 1185
Ile Glu Asn Asn Glu Lys Asp Ser Gln Arg Arg Ser Glu Lys Pro
1190 1195 1200
Leu Gln Asn Tyr Gly Phe Ser Ser Glu Met Asn Ile Glu Leu Phe
1205 1210 1215
Lys Cys Leu Met Asp Lys Ala Ala Lys Pro Pro Tyr Asp Val Lys
1220 1225 1230
Leu Ser Thr Leu Ser Lys Asn Leu Glu Glu Gly Phe Glu Lys Phe
1235 1240 1245
Lys Ala Leu Glu Leu Gly Pro Gln Val Lys Val Leu Gln Gln Ile
1250 1255 1260
Leu Asp Ile Tyr Ser Cys Asp Arg Lys Ser Gly Asp Leu Ser Val
1265 1270 1275
Leu Gly Ser Ala Arg Asn Ala Gly Arg Leu Asp Met Asn Gly Val
1280 1285 1290
Leu Ser Glu Ala Asp Gly Glu Gln Val Thr Met Ile Cys Gln Ser
1295 1300 1305
Pro Ser Gly Leu Phe Glu Lys Arg Val Pro Met Asn Glu Lys
1310 1315 1320
<210> 16
<211> 1322
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 16
Met Lys Asn Thr Asn Glu Asp Tyr Tyr Leu Gly Leu Ala Ile Gly Thr
1 5 10 15
Asp Ser Val Gly Trp Ala Val Thr Asp Lys Glu Tyr Asn Ile Leu Glu
20 25 30
Phe Arg Arg Lys Pro Met Trp Gly Ile His Leu Phe Glu Gly Gly Ser
35 40 45
Thr Ala Gln Lys Thr Arg Val Tyr Arg Thr Ser Arg Arg Arg Leu Lys
50 55 60
Arg Arg Ala Glu Arg Ile Ala Leu Leu Arg Asp Ile Phe Ser Glu Glu
65 70 75 80
Ile Gly Lys Val Asp Pro Gly Phe Phe Glu Arg Leu Asp Glu Ser Asp
85 90 95
Leu His Leu Glu Asp Arg Val Thr Ser Gln Lys Asn Ser Leu Phe Asp
100 105 110
Asp Pro Glu Phe Asn Asp Lys Asp Leu His Lys Arg Phe Pro Thr Ile
115 120 125
Tyr His Leu Arg Arg His Leu Met His Ser Asn Arg Lys Glu Asp Ile
130 135 140
Arg Leu Ile Tyr Leu Ala Ala His His Ile Ile Lys Phe Arg Gly His
145 150 155 160
Phe Leu Tyr Lys Gly Ile Gly Asp Glu Glu Ile Pro Ser Phe Glu Ile
165 170 175
Val Leu Asn Ser Leu Ile Asp Asn Leu Arg Asp Glu Tyr Gly Met Glu
180 185 190
Leu Glu Val Ser Asp Arg Asp Leu Val Lys Ala Leu Leu Ser Asp Phe
195 200 205
Ser Ile Gly Ile Arg Glu Lys Ser Arg Glu Leu Ser Ser Cys Leu Asn
210 215 220
Ala Glu Ser Glu Asn Glu Lys Ala Leu Val Asp Phe Ile Ser Gly Lys
225 230 235 240
Lys Thr Asn Met Lys Lys Leu Phe Asp Asp Glu Ala Leu Asp Lys Met
245 250 255
Ser Phe Ser Leu Arg Asp Ser Gly Phe Glu Asp Gln Leu Arg Glu Asn
260 265 270
Glu Gly Val Leu Gly Pro Glu Arg Val His Thr Leu Glu Leu Ser Arg
275 280 285
Gln Ile Phe Glu Trp Ala Arg Leu Ser Ser Ile Leu Lys Asp Ser Asp
290 295 300
Ser Ile Ser Glu Ala Lys Ile Lys Asp Tyr Asp Gln His Arg Glu Asp
305 310 315 320
Leu Arg Met Leu Lys Arg Ala Val Lys Lys Tyr Ala Pro Asp Lys Tyr
325 330 335
Ser Glu Val Phe Lys Ser Lys Glu His Thr Gly Asn Tyr Cys Ser Tyr
340 345 350
Val Tyr Val Cys Gly Lys Gly Leu Pro Asp Lys Lys Cys Ser Thr Glu
355 360 365
Glu Phe Gln Lys Tyr Leu Lys Lys Ile Leu Asp Asp Ser Gly Val Arg
370 375 380
Asp Asp Glu Glu Phe Lys Thr Leu Ile Gln Arg Leu Asp Ala Gly Ile
385 390 395 400
Leu Cys Pro Lys Gln Arg Thr Gly Glu Asn Ser Val Ile Pro Tyr Ser
405 410 415
Val His Arg Lys Glu Leu Ile Gly Ile Leu Asn Asn Ala Ala Glu His
420 425 430
Tyr Pro Ser Leu Ser Arg Lys Gly Glu Asp Gly Phe Ser Ser Ile Asp
435 440 445
Lys Ile Leu Met Leu Glu Glu Phe Arg Ile Pro Tyr Tyr Val Gly Pro
450 455 460
Leu Asp Asp Arg Ser Ser Arg Ser Trp Leu Ile Arg Asn Ser Phe Glu
465 470 475 480
Ala Ile Thr Pro Trp Asn Phe Asn Glu Ile Val Asp Glu Asp Glu Thr
485 490 495
Ser Glu Arg Phe Ile Gly Asn Leu Thr Ser Met Cys Thr Tyr Leu Gly
500 505 510
Gly Glu Lys Val Leu Pro Lys Asn Ser Leu Leu Tyr Ser Arg Phe Met
515 520 525
Leu Tyr Asn Glu Ile Asn Asn Leu Arg Val Gly Gly Glu Lys Ile Pro
530 535 540
Ala Ala Leu Lys Asn Lys Met Val Ser Glu Leu Phe Ala Asn Arg Ala
545 550 555 560
Thr Ser Ser Lys Val Thr Leu Lys Glu Leu Lys Ala Phe Leu Lys Gly
565 570 575
Glu Gly Val Leu Thr Asp Ala Asp Glu Ile Ser Gly Ile Asp Asp Gly
580 585 590
Val Lys Ser Thr Leu Arg Ser Glu Ile Leu Ile Arg Lys Ile Ile Gly
595 600 605
Asp Lys Ile Ser Asp Arg Glu Met Ala Glu Glu Ile Val Arg Ile Leu
610 615 620
Thr Val Phe Gly Asp Glu Arg Arg Arg Ser Lys Ala Lys Leu Lys Lys
625 630 635 640
Glu Phe Ser Asp Lys Leu Thr Glu Lys Glu Ile Glu Lys Leu Ser Ser
645 650 655
Leu Lys Phe Asp Gly Trp Gly Arg Leu Ser Glu Lys Phe Leu Thr Gly
660 665 670
Leu Arg Gln Glu Val Asn Gly Arg Ser Met Ser Ile Ile Glu Ile Leu
675 680 685
Glu Asp Thr Asn Tyr Asn Leu Gln Glu Thr Leu Ser Lys Tyr Ser Phe
690 695 700
Asn Glu Ile Ile Asp Ser Tyr Asn Glu Val Leu Thr Ser Gly Pro Arg
705 710 715 720
Ser Ile Ser Tyr Asp Ile Leu Lys Asp Ser Tyr Leu Ser Pro Ala Val
725 730 735
Lys Arg Gly Val Trp Arg Ala Leu Ser Val Val Lys Asp Ile Leu Lys
740 745 750
Ala Val Gly Arg Pro Pro Lys Lys Ile Phe Val Glu Thr Thr Arg Glu
755 760 765
Glu Arg Glu Lys Lys Arg Thr Glu Ser Arg Lys Asp Ala Leu Met Tyr
770 775 780
Leu Tyr Lys Ser Cys Lys Glu Thr Glu Trp Glu Lys Arg Leu Asp Ser
785 790 795 800
Val Glu Glu Ser Ser Leu Arg Asn Arg Ser Leu Tyr Leu Tyr Tyr Thr
805 810 815
Gln Leu Gly Lys Cys Met Tyr Cys Gly Lys Asn Ile Asp Ile Gly Glu
820 825 830
Leu Asn Thr Asp Leu Ala Asp Arg Asp Ala Ile Tyr Pro Gln Ser Lys
835 840 845
Thr Lys Asp Asp Ser Ile Arg Asn Asn Leu Val Leu Val Cys Arg Gly
850 855 860
Cys Asn Gln Ala Lys Gly Asp Arg Tyr Pro Leu Pro Gln Glu Trp Val
865 870 875 880
Ser Arg Met His Ala Phe Trp Thr Met Leu Lys Asp Lys Gly Tyr Ile
885 890 895
Ser Ser Glu Lys Tyr Arg Arg Leu Thr Arg Arg Gly Glu Leu Thr Glu
900 905 910
Glu Glu Phe Gly Ala Phe Ile Asn Arg Gln Leu Val Glu Thr Ser Gln
915 920 925
Ser Ala Lys Ala Val Ile Thr Val Leu Lys Asn Ala Phe Lys Asp Ser
930 935 940
Asp Ile Val Tyr Val Lys Gly Ser Asn Val Ser Asp Phe Arg Ser Ser
945 950 955 960
Tyr Asn Phe Ile Lys Cys Arg Ser Val Asn Asp Tyr His His Ala Lys
965 970 975
Asp Ala Tyr Leu Asn Ile Val Val Gly Asn Val Leu Asp Thr Lys Phe
980 985 990
Thr Lys Asn Pro Ser Tyr Val Leu Lys Asn Arg Glu Gln Tyr Asn Ile
995 1000 1005
Gly Arg Met Tyr Asp Arg Asn Val Ser Arg Phe Gly Val Asp Ala
1010 1015 1020
Trp Val Ala Gly Asp Arg Gly Ser Ile Ala Thr Val Arg Lys Tyr
1025 1030 1035
Met Arg Arg Asn Asn Ile Leu Phe Thr Arg Tyr Ala Thr Lys Ser
1040 1045 1050
Lys Gly Ala Leu Phe Lys Glu Thr Val His Arg Lys Lys Glu Gly
1055 1060 1065
Leu Phe Glu Arg Lys Lys Gly Leu Glu Thr Glu Lys Tyr Gly Gly
1070 1075 1080
Tyr Ser Asp Ile Ser Thr Ser Tyr Leu Thr Leu Leu Glu Tyr Asp
1085 1090 1095
Lys Gly Lys Lys Arg Ile Arg Ser Leu Glu Ile Val Pro Thr Tyr
1100 1105 1110
Phe Ala Asn Thr Arg Pro Lys Glu Glu Asp Val Ile Arg Phe Phe
1115 1120 1125
Ser Glu Thr Arg Gly Leu Ala Asn Val Arg Val Val Met Pro Glu
1130 1135 1140
Val Arg Met Lys Ser Leu Phe Glu Tyr Arg Gly Phe Arg Phe His
1145 1150 1155
Val Thr Gly Ser Asn Gly Lys Gly Arg Phe Trp Ile Ser Ser Ala
1160 1165 1170
Ile Gln Leu Leu Leu Pro Glu Asn Leu Tyr Ala Tyr Cys Lys Ser
1175 1180 1185
Ile Glu Asn Asn Glu Lys Asp Ser Gln Arg Arg Ser Glu Lys Pro
1190 1195 1200
Leu Gln Asn Tyr Gly Phe Ser Ser Glu Met Asn Ile Glu Leu Phe
1205 1210 1215
Lys Cys Leu Met Asp Lys Ala Ala Lys Pro Pro Tyr Asp Val Lys
1220 1225 1230
Leu Ser Thr Leu Ser Lys Asn Leu Glu Glu Gly Phe Glu Lys Phe
1235 1240 1245
Lys Ala Leu Glu Leu Gly Pro Gln Val Lys Val Leu Gln Gln Ile
1250 1255 1260
Leu Asp Ile Tyr Ser Cys Asp Arg Lys Ser Gly Asp Leu Ser Val
1265 1270 1275
Leu Gly Ser Ala Arg Asn Ala Gly Arg Leu Asp Met Asn Gly Val
1280 1285 1290
Leu Ser Glu Ala Asp Gly Glu Gln Val Thr Met Ile Cys Gln Ser
1295 1300 1305
Pro Ser Gly Leu Phe Glu Lys Arg Val Pro Met Asn Glu Lys
1310 1315 1320
<210> 17
<211> 1211
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 17
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Ala Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ala Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Val Glu Ile Leu Asn
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Ile Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Asp Asn Asn Thr Asp Lys Asp Glu Lys Glu Met Glu Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Ile Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Leu Thr Asn Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Arg Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Leu Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Phe Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Ser Thr Asn Val Gln Val
610 615 620
Asp His Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Thr Leu Glu Lys Glu Leu Ile Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Thr Asp Thr Leu Lys Pro Asn Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Thr Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Asn Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210
<210> 18
<211> 1211
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 18
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Asp Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ala Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Val Glu Ile Leu Asn
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Ile Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Asp Asn Asn Thr Asp Lys Asp Glu Lys Glu Met Glu Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Ile Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Leu Thr Asn Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Arg Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Leu Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Phe Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Ser Thr Asn Val Gln Val
610 615 620
Asp Ala Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Thr Leu Glu Lys Glu Leu Ile Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Thr Asp Thr Leu Lys Pro Asn Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Thr Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Asn Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210
<210> 19
<211> 1211
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 19
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Ala Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ala Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Val Glu Ile Leu Asn
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Ile Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Asp Asn Asn Thr Asp Lys Asp Glu Lys Glu Met Glu Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Ile Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Leu Thr Asn Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Arg Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Leu Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Phe Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Ser Thr Asn Val Gln Val
610 615 620
Asp Ala Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Thr Leu Glu Lys Glu Leu Ile Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Thr Asp Thr Leu Lys Pro Asn Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Thr Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Asn Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210
<210> 20
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 20
gtttgagagt gttgtcaaat aagagtcgga ccaatc 36
<210> 21
<211> 97
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 21
gatagacaaa tgtgtctttg acaacacaag ttcaaataag gcattgccgt aatcgttctt 60
atgaaccccg cagttggcgg gaaaccttct gttgtca 97
<210> 22
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 22
gtttgagagt gttgtcaaat aagagtcgga cc 32
<210> 23
<211> 93
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 23
gacaaatgtg tctttgacaa cacaagttca aataaggcat tgccgtaatc gttcttatga 60
accccgcagt tggcgggaaa ccttctgttg tca 93
<210> 24
<211> 13
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 24
gtttgagagt gtt 13
<210> 25
<211> 67
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 25
aacacaagtt caaataaggc attgccgtaa tcgttcttat gaaccccgca gttggcggga 60
aaccttc 67
<210> 26
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 26
gttttagtcg tctgttattt attggtaagg ttat 34
<210> 27
<211> 92
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 27
ataactttac cagtgaatat cagacggcta agataaagct ataagctgtg gggtcgcgca 60
tccccaattt cgcgcacgag cgttagctcg tt 92
<210> 28
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 28
gttttagtcg tctgttattt attggt 26
<210> 29
<211> 84
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 29
accagtgaat atcagacggc taagataaag ctataagctg tggggtcgcg catccccaat 60
ttcgcgcacg agcgttagct cgtt 84
<210> 30
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 30
gttttagtcg 10
<210> 31
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 31
cggctaagat aaagctataa gctgtggggt cgcgcatccc caatttcgcg cacgagcgtt 60
agc 63
<210> 32
<211> 14
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 32
cctgtgaata gtca 14
<210> 33
<211> 78
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 33
tataaaaaat aattatagaa ccaaactaac caattatgaa aaaaatatta ggacttgact 60
taggaaccaa ctctattg 78
<210> 34
<211> 16
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 34
cctgtgaata gtcaac 16
<210> 35
<211> 80
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 35
tatataaaaa ataattatag aaccaaacta accaattatg aaaaaaatat taggacttga 60
cttaggaacc aactctattg 80
<210> 36
<211> 12
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 36
cctgtgaata gt 12
<210> 37
<211> 56
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 37
taaaaaataa ttatagaacc aaactaacca attatgaaaa aaatattagg acttga 56
<210> 38
<211> 15
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 38
gttgtgaatt gcttt 15
<210> 39
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 39
aaagcaattc acaataagga ttattccgtt gtgaaaacat ttaaggcggt gcgaaagcat 60
cgtcct 66
<210> 40
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 40
aaagcaattc acaataagga ttattccgtt gtgaaaacat ttaaggcggt gcgaaaa 57
<210> 41
<211> 12
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 41
gttgtgaatt gc 12
<210> 42
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 42
gcaattcaca ataaggatta ttccgttgtg aaaacattta aggcggtgcg aaagcatcgt 60
c 61
<210> 43
<211> 12
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 43
gttttagagc ta 12
<210> 44
<211> 61
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 44
tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcaca gagtcggtgc 60
t 61
<210> 45
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 45
gttttagagc 10
<210> 46
<211> 59
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 46
gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcacaga gtcggtgct 59
<210> 47
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 重复
<400> 47
gttttagagt catgttgttt agaatgg 27
<210> 48
<211> 84
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> TRACR
<400> 48
ccattttaaa cgaaaaactc ctctaaaacg attgcagctt atcgtaaaaa tgaaggaacc 60
tatgattaaa gaaagccgac tgca 84
<210> 49
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物
<400> 49
ttgggtaacg ccagggtttt 20
<210> 50
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 引物
<400> 50
tgtgtggaat tgtgagcgga 20
<210> 51
<211> 1490
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 51
Met Lys Lys Ile Leu Gly Leu Ala Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Leu Ile Glu His Asn Phe Asp Lys Lys Glu Gly Arg Ile Asp Asp
20 25 30
Leu Gly Val Arg Ile Ile Pro Met Ser Ala Asp Ile Leu Gly Lys Phe
35 40 45
Asp Ala Gly Gln Ser His Ser Gln Thr Ala Glu Arg Thr Gly Tyr Arg
50 55 60
Gly Val Arg Arg Leu Tyr Gln Arg Asp Asn Leu Arg Arg Glu Arg Leu
65 70 75 80
His Arg Val Leu Asn Ile Leu Asp Phe Leu Pro Glu His Tyr Ala Glu
85 90 95
His Ile Asp Phe Glu Lys Arg Leu Gly Gln Phe Lys Glu Gly Lys Glu
100 105 110
Ile Lys Leu Asn Tyr Lys Ser Asn Lys Asp Ser Lys Phe Glu Phe Ile
115 120 125
Phe Lys Ala Ser Tyr Asn Glu Met Leu Ala Ala Phe Lys Lys Tyr Gln
130 135 140
Pro Gly Leu Phe Tyr Val Lys Ala Asn Gly Thr Glu Thr Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Ser Gln Pro
165 170 175
Leu Thr Lys Gln Glu Leu Ala Trp Ile Ile Leu Asn Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Ile Asp Asp Asp Lys Asn
195 200 205
Lys Gln Phe Val Gln Leu Lys Val Lys Glu Val Ile Asp Ser Gly Glu
210 215 220
Ala Val Lys Gly Lys Lys Leu Phe Asn Val Ile Phe Glu Asn Gly Trp
225 230 235 240
Lys Tyr Asp Lys Gln Val Val Lys Thr Glu Asp Trp Ile Gly Arg Thr
245 250 255
Lys Glu Phe Ile Val Thr Thr Lys Thr Leu Lys Ser Gly Glu Ile Lys
260 265 270
Arg Thr Tyr Lys Ala Val Asp Ser Glu Lys Asp Trp Ala Ala Ile Lys
275 280 285
Ala Lys Thr Glu Gln Asp Ile Glu Arg Ser Asn Lys Thr Val Gly Glu
290 295 300
Phe Ile Tyr Glu Ala Leu Leu Gln Asp Pro Thr Gln Lys Ile Arg Gly
305 310 315 320
Lys Leu Val Lys Thr Ile Glu Arg Lys Phe Tyr Lys Ala Glu Leu Arg
325 330 335
Glu Ile Leu Arg Lys Gln Ile Glu Leu Gln Pro Gln Leu Phe Thr Thr
340 345 350
Lys Leu Tyr Asn Ala Cys Ile Lys Glu Leu Tyr Pro Asn Asn Glu Ala
355 360 365
His Arg Asn Ser Ile Lys Asn Arg Asp Phe Leu Tyr Leu Phe Leu Asp
370 375 380
Asp Ile Ile Phe Tyr Gln Arg Pro Leu Lys Ser Gln Lys Ser Asn Ile
385 390 395 400
Ser Gly Cys His Leu Glu Gln Arg Ile Tyr Thr Lys Ile Asn Pro Val
405 410 415
Ser Gly Lys Lys Glu Glu Val Lys Gln Ala Val Lys Ala Ile Pro Lys
420 425 430
Ser His Pro Ile Phe Gln Glu Phe Arg Ile Trp Gln Trp Leu Gln Asn
435 440 445
Leu Lys Ile Tyr Asp Lys Ile Asn Thr Asp Lys Gly Glu Leu Ala Asp
450 455 460
Val Thr Asn Gln Leu Leu Pro Ser Glu Glu Ser Leu Leu Asp Leu Phe
465 470 475 480
Asp Tyr Leu Gln Thr Lys Lys Glu Leu Asp Gln Ser Gly Phe Ile Lys
485 490 495
Tyr Phe Ile Asp Lys Lys Leu Ile Asn Lys Ser Glu Lys Glu Asn Tyr
500 505 510
Arg Trp Asn Tyr Val Glu Asp Lys Lys Tyr Pro Phe Ala Glu Thr Arg
515 520 525
Ala Gln Phe Ile Ser Arg Leu Asn Lys Val Lys Asn Ile Asn Asn Ile
530 535 540
Ser Glu Phe Leu Asn Lys Lys Thr Arg Leu Gly Glu Lys Glu Ser Ser
545 550 555 560
Pro Phe Val Thr Arg Ile Glu Gln Leu Trp His Ile Ile Tyr Ser Val
565 570 575
Ser Asp Ile Asn Glu Tyr Lys Ser Ala Leu Glu Lys Phe Ala Leu Lys
580 585 590
His Asp Ile Asp Lys Glu Ser Phe Val Ala Asn Phe Ile Lys Phe Pro
595 600 605
Pro Phe Lys Ser Asp Tyr Gly Ser Tyr Ser Lys Lys Ala Leu Ser Lys
610 615 620
Leu Leu Pro Leu Met Arg Arg Gly Lys Tyr Trp Asn Glu Ser Asp Ile
625 630 635 640
Ser Asn Lys Val Lys Gln Arg Val Ser Asp Ile Met Glu Arg Val Asn
645 650 655
Ala Leu Asn Leu Lys Glu Asn Tyr Asn Ala Lys Glu Leu Ala Glu Ala
660 665 670
Leu Lys Thr Val Ser Asp Asp Asp Val Lys Lys Gln Leu Ile Lys Ser
675 680 685
Phe Val Pro Phe Lys Asp Lys Asn Pro Leu Lys Gly Leu Asn Thr Tyr
690 695 700
Gln Ala Thr Tyr Leu Val Tyr Gly Arg His Ser Glu Val Gly Asp Ile
705 710 715 720
Gln Ser Trp Lys Thr Pro Glu Asp Ile Asp Thr Tyr Leu Lys Asn Phe
725 730 735
Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr Glu
740 745 750
Thr Leu Arg Val Val Arg Asp Ile Trp Ile His Tyr Gly Lys Ser Gln
755 760 765
Leu Asn Phe Phe Asn Glu Ile His Val Glu Leu Gly Arg Glu Met Lys
770 775 780
Asn Pro Ala Asp Lys Arg Lys Gln Ile Ser Asn Arg Asn Ile Glu Asn
785 790 795 800
Glu Asn Thr Asn Asn Arg Ile Arg Glu Ile Leu Lys Asp Leu Lys Asn
805 810 815
Asp Thr Ser Ile Glu Gly Asp Ile Arg Asp Tyr Ser Pro Ser Gln Gln
820 825 830
Asp Leu Leu Lys Ile Tyr Glu Glu Gly Val Tyr Gln Asn Pro Lys Val
835 840 845
Asp Tyr Ser Lys Val Ser Glu Asp Glu Ile Thr Lys Ile Arg Arg Ser
850 855 860
Asn Ser Pro Thr Pro Lys Glu Ile Gln Arg Tyr Arg Leu Trp Leu Glu
865 870 875 880
Gln Gly Tyr Ile Ser Pro Tyr Thr Gly Lys Pro Ile Pro Leu Ser Lys
885 890 895
Leu Phe Thr His Glu Tyr Gln Ile Glu His Ile Ile Pro Gln Ser Arg
900 905 910
Tyr Phe Asp Asn Ser Leu Ser Asn Lys Ile Ile Cys Glu Ser Ala Val
915 920 925
Asn Glu Asp Lys Asp Asn Lys Thr Ala Tyr Glu Tyr Leu Lys Asn Lys
930 935 940
Ser Gly Asn Val Ile Asn Gly His Lys Leu Leu Arg Ile Glu Glu Tyr
945 950 955 960
Glu Ala His Val Asn Arg Tyr Phe Lys Asn Asn Arg Gln Lys Leu Lys
965 970 975
Asn Leu Leu Ser Glu Asp Ile Pro Glu Gly Phe Ile Asn Arg Gln Leu
980 985 990
Asn Asp Ser Arg Tyr Ile Ser Lys Leu Ile Lys Gly Leu Leu Ser Asn
995 1000 1005
Ile Val Arg Gln Glu Asn Glu Gln Glu Ala Thr Ser Lys Asn Leu
1010 1015 1020
Ile Pro Val Thr Gly Ala Val Thr Ser Lys Leu Lys Asn Asp Trp
1025 1030 1035
Gly Leu Asn Asp Lys Trp Asn Glu Leu Ile Leu Pro Arg Phe Glu
1040 1045 1050
Arg Leu Asn Gln Leu Thr Gln Thr Lys Asn Phe Thr Thr Ser Asn
1055 1060 1065
Thr Asn Gly Asn Thr Ile Pro Thr Val Pro Asp Asp Leu Leu Lys
1070 1075 1080
Gly Phe Ser Lys Lys Arg Ile Asp His Arg His His Ala Leu Asp
1085 1090 1095
Ala Leu Val Val Ala Cys Cys Thr Arg Asn His Val Gln Tyr Leu
1100 1105 1110
Asn Ala Leu Asn Ala Glu Lys Ala Asn Tyr Gly Leu Arg Lys Lys
1115 1120 1125
Leu Leu Ile Val Asn Glu Gln Gly Asp Phe Thr Lys Ile Phe Gln
1130 1135 1140
Met Pro Trp Lys Gly Phe Thr Ser Glu Ala Lys Asn Gln Leu Glu
1145 1150 1155
Lys Thr Val Ile Ser Phe Lys Gln Asn Leu Arg Val Ile Asn Lys
1160 1165 1170
Ala Asn Asn Lys Phe Trp Ser Phe Lys Asp Glu Asn Gly Asn Ile
1175 1180 1185
Asn Leu Asp Lys Asn Gly Arg Pro Val Lys Lys Leu Arg Lys Gln
1190 1195 1200
Thr Lys Gly Asp Asn Trp Ala Ile Arg Lys Ala Met His Lys Glu
1205 1210 1215
Thr Val Ser Gly Lys Ser Asn Ile Glu Thr Pro Lys Gly Lys Ile
1220 1225 1230
Ala Thr Ala Val Arg Gly Ser Leu Ala Asp Ile Lys Asn Glu Lys
1235 1240 1245
His Leu Gly Lys Ile Thr Asp Val Gln Ile Arg Glu Val Ile Leu
1250 1255 1260
Pro Asn His Leu Lys Asn Tyr Val Asp Glu Lys Gly Lys Val Lys
1265 1270 1275
Phe Asp Leu Ala Phe Asn Asp Glu Gly Ile Glu Asp Leu Asn Lys
1280 1285 1290
Asn Ile Ile Ala Leu Asn Asn Gly Lys Lys His Gln Pro Ile Arg
1295 1300 1305
Lys Val Lys Phe Phe Glu Val Gly Ser Lys Phe Ser Ile Ser Glu
1310 1315 1320
Asn Glu Asn Ser Ala Lys Ser Lys Lys Tyr Val Glu Ala Ala Lys
1325 1330 1335
Gly Thr Asn Leu Phe Phe Ala Val Tyr Trp Asp Glu Lys Lys Gln
1340 1345 1350
Lys Arg Asn Tyr Glu Thr Val Pro Leu Asn Glu Val Ile Ala His
1355 1360 1365
Gln Lys Gln Val Ala His Leu Thr Asn Asn Glu Arg Leu Pro Ile
1370 1375 1380
Gln Thr Asn Arg Lys Lys Gly Asp Phe Leu Phe Thr Leu Ser Pro
1385 1390 1395
Asn Asp Leu Val Tyr Val Pro Thr Asp Ala Glu Val Ala Asn Lys
1400 1405 1410
Gln Pro Ile Asp Phe Lys Asn Leu His Gln Asn Gln Val Asn Arg
1415 1420 1425
Ile Tyr Lys Met Val Ser Ser Ser Gly Asn Gln Cys Phe Phe Ile
1430 1435 1440
Lys Asp Lys Ile Ala Thr Ser Ile Trp Asn Lys Asn Glu Phe Ser
1445 1450 1455
Ser Leu Asn Lys Met Glu Lys Asp Ile Asp Gly Asn Met Ile Lys
1460 1465 1470
Glu Arg Cys Ile Lys Leu Asn Val Asp Arg Leu Gly Asn Ile Thr
1475 1480 1485
Lys Ala
1490
<210> 52
<211> 1490
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 52
Met Lys Lys Ile Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Leu Ile Glu His Asn Phe Asp Lys Lys Glu Gly Arg Ile Asp Asp
20 25 30
Leu Gly Val Arg Ile Ile Pro Met Ser Ala Asp Ile Leu Gly Lys Phe
35 40 45
Asp Ala Gly Gln Ser His Ser Gln Thr Ala Glu Arg Thr Gly Tyr Arg
50 55 60
Gly Val Arg Arg Leu Tyr Gln Arg Asp Asn Leu Arg Arg Glu Arg Leu
65 70 75 80
His Arg Val Leu Asn Ile Leu Asp Phe Leu Pro Glu His Tyr Ala Glu
85 90 95
His Ile Asp Phe Glu Lys Arg Leu Gly Gln Phe Lys Glu Gly Lys Glu
100 105 110
Ile Lys Leu Asn Tyr Lys Ser Asn Lys Asp Ser Lys Phe Glu Phe Ile
115 120 125
Phe Lys Ala Ser Tyr Asn Glu Met Leu Ala Ala Phe Lys Lys Tyr Gln
130 135 140
Pro Gly Leu Phe Tyr Val Lys Ala Asn Gly Thr Glu Thr Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Ser Gln Pro
165 170 175
Leu Thr Lys Gln Glu Leu Ala Trp Ile Ile Leu Asn Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Ile Asp Asp Asp Lys Asn
195 200 205
Lys Gln Phe Val Gln Leu Lys Val Lys Glu Val Ile Asp Ser Gly Glu
210 215 220
Ala Val Lys Gly Lys Lys Leu Phe Asn Val Ile Phe Glu Asn Gly Trp
225 230 235 240
Lys Tyr Asp Lys Gln Val Val Lys Thr Glu Asp Trp Ile Gly Arg Thr
245 250 255
Lys Glu Phe Ile Val Thr Thr Lys Thr Leu Lys Ser Gly Glu Ile Lys
260 265 270
Arg Thr Tyr Lys Ala Val Asp Ser Glu Lys Asp Trp Ala Ala Ile Lys
275 280 285
Ala Lys Thr Glu Gln Asp Ile Glu Arg Ser Asn Lys Thr Val Gly Glu
290 295 300
Phe Ile Tyr Glu Ala Leu Leu Gln Asp Pro Thr Gln Lys Ile Arg Gly
305 310 315 320
Lys Leu Val Lys Thr Ile Glu Arg Lys Phe Tyr Lys Ala Glu Leu Arg
325 330 335
Glu Ile Leu Arg Lys Gln Ile Glu Leu Gln Pro Gln Leu Phe Thr Thr
340 345 350
Lys Leu Tyr Asn Ala Cys Ile Lys Glu Leu Tyr Pro Asn Asn Glu Ala
355 360 365
His Arg Asn Ser Ile Lys Asn Arg Asp Phe Leu Tyr Leu Phe Leu Asp
370 375 380
Asp Ile Ile Phe Tyr Gln Arg Pro Leu Lys Ser Gln Lys Ser Asn Ile
385 390 395 400
Ser Gly Cys His Leu Glu Gln Arg Ile Tyr Thr Lys Ile Asn Pro Val
405 410 415
Ser Gly Lys Lys Glu Glu Val Lys Gln Ala Val Lys Ala Ile Pro Lys
420 425 430
Ser His Pro Ile Phe Gln Glu Phe Arg Ile Trp Gln Trp Leu Gln Asn
435 440 445
Leu Lys Ile Tyr Asp Lys Ile Asn Thr Asp Lys Gly Glu Leu Ala Asp
450 455 460
Val Thr Asn Gln Leu Leu Pro Ser Glu Glu Ser Leu Leu Asp Leu Phe
465 470 475 480
Asp Tyr Leu Gln Thr Lys Lys Glu Leu Asp Gln Ser Gly Phe Ile Lys
485 490 495
Tyr Phe Ile Asp Lys Lys Leu Ile Asn Lys Ser Glu Lys Glu Asn Tyr
500 505 510
Arg Trp Asn Tyr Val Glu Asp Lys Lys Tyr Pro Phe Ala Glu Thr Arg
515 520 525
Ala Gln Phe Ile Ser Arg Leu Asn Lys Val Lys Asn Ile Asn Asn Ile
530 535 540
Ser Glu Phe Leu Asn Lys Lys Thr Arg Leu Gly Glu Lys Glu Ser Ser
545 550 555 560
Pro Phe Val Thr Arg Ile Glu Gln Leu Trp His Ile Ile Tyr Ser Val
565 570 575
Ser Asp Ile Asn Glu Tyr Lys Ser Ala Leu Glu Lys Phe Ala Leu Lys
580 585 590
His Asp Ile Asp Lys Glu Ser Phe Val Ala Asn Phe Ile Lys Phe Pro
595 600 605
Pro Phe Lys Ser Asp Tyr Gly Ser Tyr Ser Lys Lys Ala Leu Ser Lys
610 615 620
Leu Leu Pro Leu Met Arg Arg Gly Lys Tyr Trp Asn Glu Ser Asp Ile
625 630 635 640
Ser Asn Lys Val Lys Gln Arg Val Ser Asp Ile Met Glu Arg Val Asn
645 650 655
Ala Leu Asn Leu Lys Glu Asn Tyr Asn Ala Lys Glu Leu Ala Glu Ala
660 665 670
Leu Lys Thr Val Ser Asp Asp Asp Val Lys Lys Gln Leu Ile Lys Ser
675 680 685
Phe Val Pro Phe Lys Asp Lys Asn Pro Leu Lys Gly Leu Asn Thr Tyr
690 695 700
Gln Ala Thr Tyr Leu Val Tyr Gly Arg His Ser Glu Val Gly Asp Ile
705 710 715 720
Gln Ser Trp Lys Thr Pro Glu Asp Ile Asp Thr Tyr Leu Lys Asn Phe
725 730 735
Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr Glu
740 745 750
Thr Leu Arg Val Val Arg Asp Ile Trp Ile His Tyr Gly Lys Ser Gln
755 760 765
Leu Asn Phe Phe Asn Glu Ile His Val Glu Leu Gly Arg Glu Met Lys
770 775 780
Asn Pro Ala Asp Lys Arg Lys Gln Ile Ser Asn Arg Asn Ile Glu Asn
785 790 795 800
Glu Asn Thr Asn Asn Arg Ile Arg Glu Ile Leu Lys Asp Leu Lys Asn
805 810 815
Asp Thr Ser Ile Glu Gly Asp Ile Arg Asp Tyr Ser Pro Ser Gln Gln
820 825 830
Asp Leu Leu Lys Ile Tyr Glu Glu Gly Val Tyr Gln Asn Pro Lys Val
835 840 845
Asp Tyr Ser Lys Val Ser Glu Asp Glu Ile Thr Lys Ile Arg Arg Ser
850 855 860
Asn Ser Pro Thr Pro Lys Glu Ile Gln Arg Tyr Arg Leu Trp Leu Glu
865 870 875 880
Gln Gly Tyr Ile Ser Pro Tyr Thr Gly Lys Pro Ile Pro Leu Ser Lys
885 890 895
Leu Phe Thr His Glu Tyr Gln Ile Glu Ala Ile Ile Pro Gln Ser Arg
900 905 910
Tyr Phe Asp Asn Ser Leu Ser Asn Lys Ile Ile Cys Glu Ser Ala Val
915 920 925
Asn Glu Asp Lys Asp Asn Lys Thr Ala Tyr Glu Tyr Leu Lys Asn Lys
930 935 940
Ser Gly Asn Val Ile Asn Gly His Lys Leu Leu Arg Ile Glu Glu Tyr
945 950 955 960
Glu Ala His Val Asn Arg Tyr Phe Lys Asn Asn Arg Gln Lys Leu Lys
965 970 975
Asn Leu Leu Ser Glu Asp Ile Pro Glu Gly Phe Ile Asn Arg Gln Leu
980 985 990
Asn Asp Ser Arg Tyr Ile Ser Lys Leu Ile Lys Gly Leu Leu Ser Asn
995 1000 1005
Ile Val Arg Gln Glu Asn Glu Gln Glu Ala Thr Ser Lys Asn Leu
1010 1015 1020
Ile Pro Val Thr Gly Ala Val Thr Ser Lys Leu Lys Asn Asp Trp
1025 1030 1035
Gly Leu Asn Asp Lys Trp Asn Glu Leu Ile Leu Pro Arg Phe Glu
1040 1045 1050
Arg Leu Asn Gln Leu Thr Gln Thr Lys Asn Phe Thr Thr Ser Asn
1055 1060 1065
Thr Asn Gly Asn Thr Ile Pro Thr Val Pro Asp Asp Leu Leu Lys
1070 1075 1080
Gly Phe Ser Lys Lys Arg Ile Asp His Arg His His Ala Leu Asp
1085 1090 1095
Ala Leu Val Val Ala Cys Cys Thr Arg Asn His Val Gln Tyr Leu
1100 1105 1110
Asn Ala Leu Asn Ala Glu Lys Ala Asn Tyr Gly Leu Arg Lys Lys
1115 1120 1125
Leu Leu Ile Val Asn Glu Gln Gly Asp Phe Thr Lys Ile Phe Gln
1130 1135 1140
Met Pro Trp Lys Gly Phe Thr Ser Glu Ala Lys Asn Gln Leu Glu
1145 1150 1155
Lys Thr Val Ile Ser Phe Lys Gln Asn Leu Arg Val Ile Asn Lys
1160 1165 1170
Ala Asn Asn Lys Phe Trp Ser Phe Lys Asp Glu Asn Gly Asn Ile
1175 1180 1185
Asn Leu Asp Lys Asn Gly Arg Pro Val Lys Lys Leu Arg Lys Gln
1190 1195 1200
Thr Lys Gly Asp Asn Trp Ala Ile Arg Lys Ala Met His Lys Glu
1205 1210 1215
Thr Val Ser Gly Lys Ser Asn Ile Glu Thr Pro Lys Gly Lys Ile
1220 1225 1230
Ala Thr Ala Val Arg Gly Ser Leu Ala Asp Ile Lys Asn Glu Lys
1235 1240 1245
His Leu Gly Lys Ile Thr Asp Val Gln Ile Arg Glu Val Ile Leu
1250 1255 1260
Pro Asn His Leu Lys Asn Tyr Val Asp Glu Lys Gly Lys Val Lys
1265 1270 1275
Phe Asp Leu Ala Phe Asn Asp Glu Gly Ile Glu Asp Leu Asn Lys
1280 1285 1290
Asn Ile Ile Ala Leu Asn Asn Gly Lys Lys His Gln Pro Ile Arg
1295 1300 1305
Lys Val Lys Phe Phe Glu Val Gly Ser Lys Phe Ser Ile Ser Glu
1310 1315 1320
Asn Glu Asn Ser Ala Lys Ser Lys Lys Tyr Val Glu Ala Ala Lys
1325 1330 1335
Gly Thr Asn Leu Phe Phe Ala Val Tyr Trp Asp Glu Lys Lys Gln
1340 1345 1350
Lys Arg Asn Tyr Glu Thr Val Pro Leu Asn Glu Val Ile Ala His
1355 1360 1365
Gln Lys Gln Val Ala His Leu Thr Asn Asn Glu Arg Leu Pro Ile
1370 1375 1380
Gln Thr Asn Arg Lys Lys Gly Asp Phe Leu Phe Thr Leu Ser Pro
1385 1390 1395
Asn Asp Leu Val Tyr Val Pro Thr Asp Ala Glu Val Ala Asn Lys
1400 1405 1410
Gln Pro Ile Asp Phe Lys Asn Leu His Gln Asn Gln Val Asn Arg
1415 1420 1425
Ile Tyr Lys Met Val Ser Ser Ser Gly Asn Gln Cys Phe Phe Ile
1430 1435 1440
Lys Asp Lys Ile Ala Thr Ser Ile Trp Asn Lys Asn Glu Phe Ser
1445 1450 1455
Ser Leu Asn Lys Met Glu Lys Asp Ile Asp Gly Asn Met Ile Lys
1460 1465 1470
Glu Arg Cys Ile Lys Leu Asn Val Asp Arg Leu Gly Asn Ile Thr
1475 1480 1485
Lys Ala
1490
<210> 53
<211> 1490
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 53
Met Lys Lys Ile Leu Gly Leu Ala Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Leu Ile Glu His Asn Phe Asp Lys Lys Glu Gly Arg Ile Asp Asp
20 25 30
Leu Gly Val Arg Ile Ile Pro Met Ser Ala Asp Ile Leu Gly Lys Phe
35 40 45
Asp Ala Gly Gln Ser His Ser Gln Thr Ala Glu Arg Thr Gly Tyr Arg
50 55 60
Gly Val Arg Arg Leu Tyr Gln Arg Asp Asn Leu Arg Arg Glu Arg Leu
65 70 75 80
His Arg Val Leu Asn Ile Leu Asp Phe Leu Pro Glu His Tyr Ala Glu
85 90 95
His Ile Asp Phe Glu Lys Arg Leu Gly Gln Phe Lys Glu Gly Lys Glu
100 105 110
Ile Lys Leu Asn Tyr Lys Ser Asn Lys Asp Ser Lys Phe Glu Phe Ile
115 120 125
Phe Lys Ala Ser Tyr Asn Glu Met Leu Ala Ala Phe Lys Lys Tyr Gln
130 135 140
Pro Gly Leu Phe Tyr Val Lys Ala Asn Gly Thr Glu Thr Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Ser Gln Pro
165 170 175
Leu Thr Lys Gln Glu Leu Ala Trp Ile Ile Leu Asn Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Ile Asp Asp Asp Lys Asn
195 200 205
Lys Gln Phe Val Gln Leu Lys Val Lys Glu Val Ile Asp Ser Gly Glu
210 215 220
Ala Val Lys Gly Lys Lys Leu Phe Asn Val Ile Phe Glu Asn Gly Trp
225 230 235 240
Lys Tyr Asp Lys Gln Val Val Lys Thr Glu Asp Trp Ile Gly Arg Thr
245 250 255
Lys Glu Phe Ile Val Thr Thr Lys Thr Leu Lys Ser Gly Glu Ile Lys
260 265 270
Arg Thr Tyr Lys Ala Val Asp Ser Glu Lys Asp Trp Ala Ala Ile Lys
275 280 285
Ala Lys Thr Glu Gln Asp Ile Glu Arg Ser Asn Lys Thr Val Gly Glu
290 295 300
Phe Ile Tyr Glu Ala Leu Leu Gln Asp Pro Thr Gln Lys Ile Arg Gly
305 310 315 320
Lys Leu Val Lys Thr Ile Glu Arg Lys Phe Tyr Lys Ala Glu Leu Arg
325 330 335
Glu Ile Leu Arg Lys Gln Ile Glu Leu Gln Pro Gln Leu Phe Thr Thr
340 345 350
Lys Leu Tyr Asn Ala Cys Ile Lys Glu Leu Tyr Pro Asn Asn Glu Ala
355 360 365
His Arg Asn Ser Ile Lys Asn Arg Asp Phe Leu Tyr Leu Phe Leu Asp
370 375 380
Asp Ile Ile Phe Tyr Gln Arg Pro Leu Lys Ser Gln Lys Ser Asn Ile
385 390 395 400
Ser Gly Cys His Leu Glu Gln Arg Ile Tyr Thr Lys Ile Asn Pro Val
405 410 415
Ser Gly Lys Lys Glu Glu Val Lys Gln Ala Val Lys Ala Ile Pro Lys
420 425 430
Ser His Pro Ile Phe Gln Glu Phe Arg Ile Trp Gln Trp Leu Gln Asn
435 440 445
Leu Lys Ile Tyr Asp Lys Ile Asn Thr Asp Lys Gly Glu Leu Ala Asp
450 455 460
Val Thr Asn Gln Leu Leu Pro Ser Glu Glu Ser Leu Leu Asp Leu Phe
465 470 475 480
Asp Tyr Leu Gln Thr Lys Lys Glu Leu Asp Gln Ser Gly Phe Ile Lys
485 490 495
Tyr Phe Ile Asp Lys Lys Leu Ile Asn Lys Ser Glu Lys Glu Asn Tyr
500 505 510
Arg Trp Asn Tyr Val Glu Asp Lys Lys Tyr Pro Phe Ala Glu Thr Arg
515 520 525
Ala Gln Phe Ile Ser Arg Leu Asn Lys Val Lys Asn Ile Asn Asn Ile
530 535 540
Ser Glu Phe Leu Asn Lys Lys Thr Arg Leu Gly Glu Lys Glu Ser Ser
545 550 555 560
Pro Phe Val Thr Arg Ile Glu Gln Leu Trp His Ile Ile Tyr Ser Val
565 570 575
Ser Asp Ile Asn Glu Tyr Lys Ser Ala Leu Glu Lys Phe Ala Leu Lys
580 585 590
His Asp Ile Asp Lys Glu Ser Phe Val Ala Asn Phe Ile Lys Phe Pro
595 600 605
Pro Phe Lys Ser Asp Tyr Gly Ser Tyr Ser Lys Lys Ala Leu Ser Lys
610 615 620
Leu Leu Pro Leu Met Arg Arg Gly Lys Tyr Trp Asn Glu Ser Asp Ile
625 630 635 640
Ser Asn Lys Val Lys Gln Arg Val Ser Asp Ile Met Glu Arg Val Asn
645 650 655
Ala Leu Asn Leu Lys Glu Asn Tyr Asn Ala Lys Glu Leu Ala Glu Ala
660 665 670
Leu Lys Thr Val Ser Asp Asp Asp Val Lys Lys Gln Leu Ile Lys Ser
675 680 685
Phe Val Pro Phe Lys Asp Lys Asn Pro Leu Lys Gly Leu Asn Thr Tyr
690 695 700
Gln Ala Thr Tyr Leu Val Tyr Gly Arg His Ser Glu Val Gly Asp Ile
705 710 715 720
Gln Ser Trp Lys Thr Pro Glu Asp Ile Asp Thr Tyr Leu Lys Asn Phe
725 730 735
Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr Glu
740 745 750
Thr Leu Arg Val Val Arg Asp Ile Trp Ile His Tyr Gly Lys Ser Gln
755 760 765
Leu Asn Phe Phe Asn Glu Ile His Val Glu Leu Gly Arg Glu Met Lys
770 775 780
Asn Pro Ala Asp Lys Arg Lys Gln Ile Ser Asn Arg Asn Ile Glu Asn
785 790 795 800
Glu Asn Thr Asn Asn Arg Ile Arg Glu Ile Leu Lys Asp Leu Lys Asn
805 810 815
Asp Thr Ser Ile Glu Gly Asp Ile Arg Asp Tyr Ser Pro Ser Gln Gln
820 825 830
Asp Leu Leu Lys Ile Tyr Glu Glu Gly Val Tyr Gln Asn Pro Lys Val
835 840 845
Asp Tyr Ser Lys Val Ser Glu Asp Glu Ile Thr Lys Ile Arg Arg Ser
850 855 860
Asn Ser Pro Thr Pro Lys Glu Ile Gln Arg Tyr Arg Leu Trp Leu Glu
865 870 875 880
Gln Gly Tyr Ile Ser Pro Tyr Thr Gly Lys Pro Ile Pro Leu Ser Lys
885 890 895
Leu Phe Thr His Glu Tyr Gln Ile Glu Ala Ile Ile Pro Gln Ser Arg
900 905 910
Tyr Phe Asp Asn Ser Leu Ser Asn Lys Ile Ile Cys Glu Ser Ala Val
915 920 925
Asn Glu Asp Lys Asp Asn Lys Thr Ala Tyr Glu Tyr Leu Lys Asn Lys
930 935 940
Ser Gly Asn Val Ile Asn Gly His Lys Leu Leu Arg Ile Glu Glu Tyr
945 950 955 960
Glu Ala His Val Asn Arg Tyr Phe Lys Asn Asn Arg Gln Lys Leu Lys
965 970 975
Asn Leu Leu Ser Glu Asp Ile Pro Glu Gly Phe Ile Asn Arg Gln Leu
980 985 990
Asn Asp Ser Arg Tyr Ile Ser Lys Leu Ile Lys Gly Leu Leu Ser Asn
995 1000 1005
Ile Val Arg Gln Glu Asn Glu Gln Glu Ala Thr Ser Lys Asn Leu
1010 1015 1020
Ile Pro Val Thr Gly Ala Val Thr Ser Lys Leu Lys Asn Asp Trp
1025 1030 1035
Gly Leu Asn Asp Lys Trp Asn Glu Leu Ile Leu Pro Arg Phe Glu
1040 1045 1050
Arg Leu Asn Gln Leu Thr Gln Thr Lys Asn Phe Thr Thr Ser Asn
1055 1060 1065
Thr Asn Gly Asn Thr Ile Pro Thr Val Pro Asp Asp Leu Leu Lys
1070 1075 1080
Gly Phe Ser Lys Lys Arg Ile Asp His Arg His His Ala Leu Asp
1085 1090 1095
Ala Leu Val Val Ala Cys Cys Thr Arg Asn His Val Gln Tyr Leu
1100 1105 1110
Asn Ala Leu Asn Ala Glu Lys Ala Asn Tyr Gly Leu Arg Lys Lys
1115 1120 1125
Leu Leu Ile Val Asn Glu Gln Gly Asp Phe Thr Lys Ile Phe Gln
1130 1135 1140
Met Pro Trp Lys Gly Phe Thr Ser Glu Ala Lys Asn Gln Leu Glu
1145 1150 1155
Lys Thr Val Ile Ser Phe Lys Gln Asn Leu Arg Val Ile Asn Lys
1160 1165 1170
Ala Asn Asn Lys Phe Trp Ser Phe Lys Asp Glu Asn Gly Asn Ile
1175 1180 1185
Asn Leu Asp Lys Asn Gly Arg Pro Val Lys Lys Leu Arg Lys Gln
1190 1195 1200
Thr Lys Gly Asp Asn Trp Ala Ile Arg Lys Ala Met His Lys Glu
1205 1210 1215
Thr Val Ser Gly Lys Ser Asn Ile Glu Thr Pro Lys Gly Lys Ile
1220 1225 1230
Ala Thr Ala Val Arg Gly Ser Leu Ala Asp Ile Lys Asn Glu Lys
1235 1240 1245
His Leu Gly Lys Ile Thr Asp Val Gln Ile Arg Glu Val Ile Leu
1250 1255 1260
Pro Asn His Leu Lys Asn Tyr Val Asp Glu Lys Gly Lys Val Lys
1265 1270 1275
Phe Asp Leu Ala Phe Asn Asp Glu Gly Ile Glu Asp Leu Asn Lys
1280 1285 1290
Asn Ile Ile Ala Leu Asn Asn Gly Lys Lys His Gln Pro Ile Arg
1295 1300 1305
Lys Val Lys Phe Phe Glu Val Gly Ser Lys Phe Ser Ile Ser Glu
1310 1315 1320
Asn Glu Asn Ser Ala Lys Ser Lys Lys Tyr Val Glu Ala Ala Lys
1325 1330 1335
Gly Thr Asn Leu Phe Phe Ala Val Tyr Trp Asp Glu Lys Lys Gln
1340 1345 1350
Lys Arg Asn Tyr Glu Thr Val Pro Leu Asn Glu Val Ile Ala His
1355 1360 1365
Gln Lys Gln Val Ala His Leu Thr Asn Asn Glu Arg Leu Pro Ile
1370 1375 1380
Gln Thr Asn Arg Lys Lys Gly Asp Phe Leu Phe Thr Leu Ser Pro
1385 1390 1395
Asn Asp Leu Val Tyr Val Pro Thr Asp Ala Glu Val Ala Asn Lys
1400 1405 1410
Gln Pro Ile Asp Phe Lys Asn Leu His Gln Asn Gln Val Asn Arg
1415 1420 1425
Ile Tyr Lys Met Val Ser Ser Ser Gly Asn Gln Cys Phe Phe Ile
1430 1435 1440
Lys Asp Lys Ile Ala Thr Ser Ile Trp Asn Lys Asn Glu Phe Ser
1445 1450 1455
Ser Leu Asn Lys Met Glu Lys Asp Ile Asp Gly Asn Met Ile Lys
1460 1465 1470
Glu Arg Cys Ile Lys Leu Asn Val Asp Arg Leu Gly Asn Ile Thr
1475 1480 1485
Lys Ala
1490
<210> 54
<211> 1511
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 54
Met Lys Lys Ile Leu Gly Leu Ala Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Val Val Asn Ala Asp Ala Ile Thr Arg Asn Asp Gly Ser Arg Tyr
20 25 30
Leu Lys Pro Asn Ser Ile Ser Ala Ala Gly Ser Arg Ile Ile Pro Met
35 40 45
Ser Ala Asp Val Leu Gly Asn Phe Glu Ser Gly Ile Thr Val Ser Gln
50 55 60
Thr Lys Asp Arg Thr Asp Lys Arg Met Ala Arg Arg Leu His Glu Arg
65 70 75 80
Ala Leu Leu Arg Arg Glu Arg Leu Leu Arg Ile Leu Ser Leu Met Asp
85 90 95
Phe Leu Pro Lys His Phe Ala Ser Lys Ile Asn Arg Tyr Gly Lys Phe
100 105 110
Thr Asp Asp Ser Glu Pro Lys Leu Ala Trp Arg Lys Asn Thr Glu Gly
115 120 125
Lys Tyr Glu Phe Ile Phe Gln Asp Ala Phe Asn Glu Met Leu Ala Glu
130 135 140
Phe Lys Asp Lys Gln Pro Glu Ile Val Lys Glu Gly Lys Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Glu Lys Ala
165 170 175
Leu Ser Lys Glu Glu Leu Ser Trp Leu Leu Leu Gln Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Glu Asp Ile Pro Gln Asp
195 200 205
Lys Lys Ile Glu Tyr Leu Ala Gln Lys Val Val Lys Val Glu Ala Thr
210 215 220
Asp Gln Lys Lys Gly Asp Asp Ile Trp Tyr Asn Val Tyr Leu Glu Asn
225 230 235 240
Gly Met Ile Tyr Arg Arg Thr Ser Lys Ala Pro Leu Asp Trp Glu Gly
245 250 255
Lys Ile Lys Glu Phe Ile Val Thr Thr Asp Leu Glu Lys Asp Gly Thr
260 265 270
Pro Lys Lys Asp Lys Glu Gly Asn Ile Lys Arg Ser Phe Arg Ala Pro
275 280 285
Gln Glu Asp Asp Trp Thr Leu Leu Lys Lys Lys Thr Glu Ala Asp Ile
290 295 300
Glu Lys Ser Thr Lys Thr Val Gly Cys Tyr Ile Tyr Asp Ser Leu Leu
305 310 315 320
Asn Asn Pro Lys Gln Lys Ile Ile Gly Lys Leu Val Arg Thr Val Glu
325 330 335
Arg Lys Phe Tyr Lys Glu Glu Leu Thr Gln Ile Leu Lys Lys Gln Val
340 345 350
Glu Leu Ile Pro Glu Leu Arg Asn Asp Asn Leu Tyr Lys Gln Cys Ile
355 360 365
Glu Glu Leu Tyr Pro Ile Asn Glu Ala His Arg Asn Thr Ile Ala Lys
370 375 380
Thr Asp Phe Ala Asn Leu Phe Ile Asn Asp Ile Leu Phe Tyr Gln Arg
385 390 395 400
Pro Leu Lys Ser Lys Lys Ser Gln Ile Asp Asn Cys Pro Tyr Glu Glu
405 410 415
His Ile Phe Ile Asp Ser Lys Thr Gly Glu Lys Lys Lys Val Pro Val
420 425 430
Lys Cys Ile Thr Lys Ser Asn Pro Leu Phe Gln Glu Phe Arg Leu Trp
435 440 445
Gln Phe Ile Gln Asn Leu Arg Ile Tyr Gln Arg Glu Lys Glu Ile Asp
450 455 460
Gly Lys Leu Ser Thr Asp Val Asp Ile Thr Ser Glu Cys Leu Lys Ser
465 470 475 480
Glu Glu Asp Tyr Val Arg Leu Phe Asp Trp Leu Asn Asp Arg Glu Ser
485 490 495
Ile Glu Gln Glu Glu Leu Leu Lys Tyr Leu Phe Asn Thr Lys Lys Ser
500 505 510
Lys Asn Lys Glu Asn Pro Tyr Arg Trp Asn Tyr Val Glu Asp Lys Val
515 520 525
Tyr Pro Cys Asn Glu Thr Arg Ala Thr Ile Leu Lys Gly Leu Ser Lys
530 535 540
Cys Gly Ile Asn Ala Ser Val Leu Ser Ser Glu Met Glu Met Ala Leu
545 550 555 560
Trp His Ile Leu Tyr Ser Val Glu Asp Lys Lys Glu Ile Glu Thr Ala
565 570 575
Leu Thr His Phe Ala Gln Lys Gln Gly Trp Asn Gly Glu Phe Ala Ile
580 585 590
Val Phe Ser Lys Leu Lys Pro Phe Lys Lys Asp Tyr Gly Ser Tyr Ser
595 600 605
Glu Lys Ala Ile Lys Lys Leu Leu Ser Leu Met Arg Met Gly Lys Tyr
610 615 620
Trp Asn Gln Asp Asn Ile Asp Lys Asn Thr Leu Asp Arg Ile Asp Lys
625 630 635 640
Ile Ile Asn Gly Glu Tyr Asp Glu Lys Ile Ser Asn Arg Val Arg Asp
645 650 655
Asn Ala Ile Asn Leu Lys Asp Ile Ser Asp Phe Arg Gly Leu Pro Val
660 665 670
Trp Leu Ala Cys Tyr Ile Val Tyr Asp Arg His Ser Glu Ala Lys Asp
675 680 685
Cys Thr Lys Trp Asn Thr Pro Glu Glu Ile Asp Ser Tyr Leu Lys Lys
690 695 700
Phe Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr
705 710 715 720
Glu Thr Leu Arg Thr Val Arg Asp Ile Trp Lys Gln Glu Gly Gln Ile
725 730 735
Asp Glu Ile His Leu Glu Leu Gly Arg Asp Leu Lys Asn Pro Ala Asp
740 745 750
Lys Arg Lys Lys Met Ser Glu Asn Ile Leu Lys Asn Glu Asn Thr Asn
755 760 765
Leu Arg Ile Lys Ala Met Leu Met Glu Phe Met Asn Pro Gly Met Gly
770 775 780
Ile Glu Asn Val Arg Pro Tyr Ser Pro Ser Gln Gln Asp Ile Leu Arg
785 790 795 800
Ile Tyr Glu Glu Asn Ala Leu Glu Asn Leu Thr Lys Asp Asp Glu Glu
805 810 815
Phe Asp Phe Ile Ser Lys Ile Ser Lys Gln Ala Gln Pro Thr Lys Ser
820 825 830
Asp Ile Val Arg Tyr Lys Cys Trp Leu Glu Gln Lys Tyr Arg Ser Pro
835 840 845
Tyr Thr Gly Lys Thr Ile Ser Leu Ser Lys Leu Phe Thr Ser Ala Tyr
850 855 860
Glu Ile Glu His Ile Ile Pro Gln Ser Arg Tyr Phe Asp Asp Ser Phe
865 870 875 880
Ser Asn Lys Val Ile Cys Glu Ala Glu Val Asn Lys Leu Lys Asp Arg
885 890 895
Gln Leu Gly His Glu Phe Ile Glu Glu His His Gly Glu Lys Val Gln
900 905 910
Leu Ser Gln Gly Glu Val Val Glu Ile Leu Ser Val Asp Ala Tyr Glu
915 920 925
Lys Phe Val Lys Glu Asn Tyr Ala Asn Asn Arg Val Lys Met Lys Lys
930 935 940
Leu Leu Met Glu Asn Ile Pro Asp Glu Phe Ile Glu Arg Gln Leu Asn
945 950 955 960
Asp Ser Arg Tyr Ile Ser Lys Val Val Lys Gly Leu Leu Ser Asn Ile
965 970 975
Val Arg Glu Lys Ile Asp Asp Glu Asn Tyr Glu Pro Glu Ala Val Ser
980 985 990
Lys Asn Leu Ile Ser Cys Asn Gly Ala Val Thr Asp Arg Leu Lys Lys
995 1000 1005
Asp Trp Gly Met Asn Asp Val Trp Asn Ser Ile Ile Leu Pro Arg
1010 1015 1020
Phe Ile Arg Met Asn Gln Ile Thr Gly Lys Asp Cys Phe Thr Thr
1025 1030 1035
Thr Asn Ala Glu Gly His Leu Ile Pro Gln Met Pro Leu Glu Leu
1040 1045 1050
Gln Lys Gly Phe Asn Lys Lys Arg Ile Asp His Arg His His Ala
1055 1060 1065
Met Asp Ala Ile Val Ile Ala Cys Thr Thr Arg Asp His Val Asn
1070 1075 1080
Leu Leu Asn Asn Glu Ala Ala His Ser Lys Phe Asn Ala Thr Arg
1085 1090 1095
Tyr Gln Leu Gln Arg Lys Leu Arg Cys Phe Glu Lys Ala Met Ile
1100 1105 1110
Asp Gly Lys Glu Arg Glu Val Ala Lys Glu Phe Leu Lys Pro Trp
1115 1120 1125
Asp Ser Phe Thr Met Asp Ser Lys Asn Ile Leu Glu Asn Ile Ile
1130 1135 1140
Val Ser Phe Lys Gln Asn Gln Arg Val Ile Asn Lys Thr Thr Asn
1145 1150 1155
Thr Phe Gln His Phe Asp Glu Asn Gly Lys Lys Thr Phe Val Lys
1160 1165 1170
Gln Gly Lys Gly Asn Ser Trp Ala Ile Arg Lys Pro Met His Lys
1175 1180 1185
Asp Thr Val Phe Gly Glu Ile Asn Leu Arg Lys Val Lys Ser Val
1190 1195 1200
Ser Leu Ser Asp Ala Ile Lys Val Pro Glu Arg Ile Leu Asn Lys
1205 1210 1215
Arg Ile Lys Glu Lys Ile Thr Glu Leu Lys Asn Asn Lys Val Asp
1220 1225 1230
Ala Lys Asn Ile Lys Lys Tyr Ile Glu Glu Tyr His Ile Gly Gly
1235 1240 1245
Tyr Gly Ile Asp Thr Ser Lys Ile Asp Val Phe Tyr Phe Thr Lys
1250 1255 1260
Glu Thr Lys Glu Arg Phe Phe Ala Thr Arg Lys Ser Leu Asp Ser
1265 1270 1275
Ser Phe Asn Gln Ala Lys Ile Glu Asp Ser Ile Ala Asp Ser Gly
1280 1285 1290
Ile Gln Lys Ile Leu Leu Ala His Leu Lys Ser Lys Asn Gly Asp
1295 1300 1305
Ala Glu Gln Ala Phe Ser Pro Asp Gly Ile Asp Glu Met Asn Lys
1310 1315 1320
Asn Ile Val Glu Leu Asn Asn Gly Lys Phe His Gln Pro Ile Leu
1325 1330 1335
Lys Val Arg Val Tyr Glu Lys Ala Asp Lys Phe Ala Val Gly Gln
1340 1345 1350
Lys Gly Asn Lys Lys Val Lys Phe Val Glu Ala Ala Lys Gly Thr
1355 1360 1365
Asn Leu Phe Phe Ala Val Phe Glu Lys Asp Gly Lys Arg Ser Tyr
1370 1375 1380
Leu Thr Ile Pro Leu Asn Val Met Ile Asp Cys Gln Lys Lys Tyr
1385 1390 1395
Gly Asn Gln Trp Lys Gln Asn Ile Glu Ser Tyr Leu Lys Glu Lys
1400 1405 1410
Asp Leu Val Glu Lys Asp Val Gln Leu Leu Phe Ile Leu Ser Pro
1415 1420 1425
Asn Asp Leu Val Tyr Leu Pro Thr Glu Asn Glu Leu Lys Lys Gly
1430 1435 1440
Ile Thr Asn Pro Asp Lys Asp Gln Ile Tyr Lys Phe Val Ser Cys
1445 1450 1455
Thr Ser Asn Glu Ala His Phe Ile Pro Ser Phe Val Ala Asn Pro
1460 1465 1470
Ile Val Gln Thr Thr Glu Leu Gly Ser Asn Asn Lys Ala Gln Arg
1475 1480 1485
Ala Trp Asn Asn Lys Met Ile Lys Glu Ile Cys Ile Pro Ile Glu
1490 1495 1500
Val Asp Arg Leu Gly Asn Ile Lys
1505 1510
<210> 55
<211> 1511
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 55
Met Lys Lys Ile Leu Gly Leu Asp Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Val Val Asn Ala Asp Ala Ile Thr Arg Asn Asp Gly Ser Arg Tyr
20 25 30
Leu Lys Pro Asn Ser Ile Ser Ala Ala Gly Ser Arg Ile Ile Pro Met
35 40 45
Ser Ala Asp Val Leu Gly Asn Phe Glu Ser Gly Ile Thr Val Ser Gln
50 55 60
Thr Lys Asp Arg Thr Asp Lys Arg Met Ala Arg Arg Leu His Glu Arg
65 70 75 80
Ala Leu Leu Arg Arg Glu Arg Leu Leu Arg Ile Leu Ser Leu Met Asp
85 90 95
Phe Leu Pro Lys His Phe Ala Ser Lys Ile Asn Arg Tyr Gly Lys Phe
100 105 110
Thr Asp Asp Ser Glu Pro Lys Leu Ala Trp Arg Lys Asn Thr Glu Gly
115 120 125
Lys Tyr Glu Phe Ile Phe Gln Asp Ala Phe Asn Glu Met Leu Ala Glu
130 135 140
Phe Lys Asp Lys Gln Pro Glu Ile Val Lys Glu Gly Lys Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Glu Lys Ala
165 170 175
Leu Ser Lys Glu Glu Leu Ser Trp Leu Leu Leu Gln Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Glu Asp Ile Pro Gln Asp
195 200 205
Lys Lys Ile Glu Tyr Leu Ala Gln Lys Val Val Lys Val Glu Ala Thr
210 215 220
Asp Gln Lys Lys Gly Asp Asp Ile Trp Tyr Asn Val Tyr Leu Glu Asn
225 230 235 240
Gly Met Ile Tyr Arg Arg Thr Ser Lys Ala Pro Leu Asp Trp Glu Gly
245 250 255
Lys Ile Lys Glu Phe Ile Val Thr Thr Asp Leu Glu Lys Asp Gly Thr
260 265 270
Pro Lys Lys Asp Lys Glu Gly Asn Ile Lys Arg Ser Phe Arg Ala Pro
275 280 285
Gln Glu Asp Asp Trp Thr Leu Leu Lys Lys Lys Thr Glu Ala Asp Ile
290 295 300
Glu Lys Ser Thr Lys Thr Val Gly Cys Tyr Ile Tyr Asp Ser Leu Leu
305 310 315 320
Asn Asn Pro Lys Gln Lys Ile Ile Gly Lys Leu Val Arg Thr Val Glu
325 330 335
Arg Lys Phe Tyr Lys Glu Glu Leu Thr Gln Ile Leu Lys Lys Gln Val
340 345 350
Glu Leu Ile Pro Glu Leu Arg Asn Asp Asn Leu Tyr Lys Gln Cys Ile
355 360 365
Glu Glu Leu Tyr Pro Ile Asn Glu Ala His Arg Asn Thr Ile Ala Lys
370 375 380
Thr Asp Phe Ala Asn Leu Phe Ile Asn Asp Ile Leu Phe Tyr Gln Arg
385 390 395 400
Pro Leu Lys Ser Lys Lys Ser Gln Ile Asp Asn Cys Pro Tyr Glu Glu
405 410 415
His Ile Phe Ile Asp Ser Lys Thr Gly Glu Lys Lys Lys Val Pro Val
420 425 430
Lys Cys Ile Thr Lys Ser Asn Pro Leu Phe Gln Glu Phe Arg Leu Trp
435 440 445
Gln Phe Ile Gln Asn Leu Arg Ile Tyr Gln Arg Glu Lys Glu Ile Asp
450 455 460
Gly Lys Leu Ser Thr Asp Val Asp Ile Thr Ser Glu Cys Leu Lys Ser
465 470 475 480
Glu Glu Asp Tyr Val Arg Leu Phe Asp Trp Leu Asn Asp Arg Glu Ser
485 490 495
Ile Glu Gln Glu Glu Leu Leu Lys Tyr Leu Phe Asn Thr Lys Lys Ser
500 505 510
Lys Asn Lys Glu Asn Pro Tyr Arg Trp Asn Tyr Val Glu Asp Lys Val
515 520 525
Tyr Pro Cys Asn Glu Thr Arg Ala Thr Ile Leu Lys Gly Leu Ser Lys
530 535 540
Cys Gly Ile Asn Ala Ser Val Leu Ser Ser Glu Met Glu Met Ala Leu
545 550 555 560
Trp His Ile Leu Tyr Ser Val Glu Asp Lys Lys Glu Ile Glu Thr Ala
565 570 575
Leu Thr His Phe Ala Gln Lys Gln Gly Trp Asn Gly Glu Phe Ala Ile
580 585 590
Val Phe Ser Lys Leu Lys Pro Phe Lys Lys Asp Tyr Gly Ser Tyr Ser
595 600 605
Glu Lys Ala Ile Lys Lys Leu Leu Ser Leu Met Arg Met Gly Lys Tyr
610 615 620
Trp Asn Gln Asp Asn Ile Asp Lys Asn Thr Leu Asp Arg Ile Asp Lys
625 630 635 640
Ile Ile Asn Gly Glu Tyr Asp Glu Lys Ile Ser Asn Arg Val Arg Asp
645 650 655
Asn Ala Ile Asn Leu Lys Asp Ile Ser Asp Phe Arg Gly Leu Pro Val
660 665 670
Trp Leu Ala Cys Tyr Ile Val Tyr Asp Arg His Ser Glu Ala Lys Asp
675 680 685
Cys Thr Lys Trp Asn Thr Pro Glu Glu Ile Asp Ser Tyr Leu Lys Lys
690 695 700
Phe Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr
705 710 715 720
Glu Thr Leu Arg Thr Val Arg Asp Ile Trp Lys Gln Glu Gly Gln Ile
725 730 735
Asp Glu Ile His Leu Glu Leu Gly Arg Asp Leu Lys Asn Pro Ala Asp
740 745 750
Lys Arg Lys Lys Met Ser Glu Asn Ile Leu Lys Asn Glu Asn Thr Asn
755 760 765
Leu Arg Ile Lys Ala Met Leu Met Glu Phe Met Asn Pro Gly Met Gly
770 775 780
Ile Glu Asn Val Arg Pro Tyr Ser Pro Ser Gln Gln Asp Ile Leu Arg
785 790 795 800
Ile Tyr Glu Glu Asn Ala Leu Glu Asn Leu Thr Lys Asp Asp Glu Glu
805 810 815
Phe Asp Phe Ile Ser Lys Ile Ser Lys Gln Ala Gln Pro Thr Lys Ser
820 825 830
Asp Ile Val Arg Tyr Lys Cys Trp Leu Glu Gln Lys Tyr Arg Ser Pro
835 840 845
Tyr Thr Gly Lys Thr Ile Ser Leu Ser Lys Leu Phe Thr Ser Ala Tyr
850 855 860
Glu Ile Glu Ala Ile Ile Pro Gln Ser Arg Tyr Phe Asp Asp Ser Phe
865 870 875 880
Ser Asn Lys Val Ile Cys Glu Ala Glu Val Asn Lys Leu Lys Asp Arg
885 890 895
Gln Leu Gly His Glu Phe Ile Glu Glu His His Gly Glu Lys Val Gln
900 905 910
Leu Ser Gln Gly Glu Val Val Glu Ile Leu Ser Val Asp Ala Tyr Glu
915 920 925
Lys Phe Val Lys Glu Asn Tyr Ala Asn Asn Arg Val Lys Met Lys Lys
930 935 940
Leu Leu Met Glu Asn Ile Pro Asp Glu Phe Ile Glu Arg Gln Leu Asn
945 950 955 960
Asp Ser Arg Tyr Ile Ser Lys Val Val Lys Gly Leu Leu Ser Asn Ile
965 970 975
Val Arg Glu Lys Ile Asp Asp Glu Asn Tyr Glu Pro Glu Ala Val Ser
980 985 990
Lys Asn Leu Ile Ser Cys Asn Gly Ala Val Thr Asp Arg Leu Lys Lys
995 1000 1005
Asp Trp Gly Met Asn Asp Val Trp Asn Ser Ile Ile Leu Pro Arg
1010 1015 1020
Phe Ile Arg Met Asn Gln Ile Thr Gly Lys Asp Cys Phe Thr Thr
1025 1030 1035
Thr Asn Ala Glu Gly His Leu Ile Pro Gln Met Pro Leu Glu Leu
1040 1045 1050
Gln Lys Gly Phe Asn Lys Lys Arg Ile Asp His Arg His His Ala
1055 1060 1065
Met Asp Ala Ile Val Ile Ala Cys Thr Thr Arg Asp His Val Asn
1070 1075 1080
Leu Leu Asn Asn Glu Ala Ala His Ser Lys Phe Asn Ala Thr Arg
1085 1090 1095
Tyr Gln Leu Gln Arg Lys Leu Arg Cys Phe Glu Lys Ala Met Ile
1100 1105 1110
Asp Gly Lys Glu Arg Glu Val Ala Lys Glu Phe Leu Lys Pro Trp
1115 1120 1125
Asp Ser Phe Thr Met Asp Ser Lys Asn Ile Leu Glu Asn Ile Ile
1130 1135 1140
Val Ser Phe Lys Gln Asn Gln Arg Val Ile Asn Lys Thr Thr Asn
1145 1150 1155
Thr Phe Gln His Phe Asp Glu Asn Gly Lys Lys Thr Phe Val Lys
1160 1165 1170
Gln Gly Lys Gly Asn Ser Trp Ala Ile Arg Lys Pro Met His Lys
1175 1180 1185
Asp Thr Val Phe Gly Glu Ile Asn Leu Arg Lys Val Lys Ser Val
1190 1195 1200
Ser Leu Ser Asp Ala Ile Lys Val Pro Glu Arg Ile Leu Asn Lys
1205 1210 1215
Arg Ile Lys Glu Lys Ile Thr Glu Leu Lys Asn Asn Lys Val Asp
1220 1225 1230
Ala Lys Asn Ile Lys Lys Tyr Ile Glu Glu Tyr His Ile Gly Gly
1235 1240 1245
Tyr Gly Ile Asp Thr Ser Lys Ile Asp Val Phe Tyr Phe Thr Lys
1250 1255 1260
Glu Thr Lys Glu Arg Phe Phe Ala Thr Arg Lys Ser Leu Asp Ser
1265 1270 1275
Ser Phe Asn Gln Ala Lys Ile Glu Asp Ser Ile Ala Asp Ser Gly
1280 1285 1290
Ile Gln Lys Ile Leu Leu Ala His Leu Lys Ser Lys Asn Gly Asp
1295 1300 1305
Ala Glu Gln Ala Phe Ser Pro Asp Gly Ile Asp Glu Met Asn Lys
1310 1315 1320
Asn Ile Val Glu Leu Asn Asn Gly Lys Phe His Gln Pro Ile Leu
1325 1330 1335
Lys Val Arg Val Tyr Glu Lys Ala Asp Lys Phe Ala Val Gly Gln
1340 1345 1350
Lys Gly Asn Lys Lys Val Lys Phe Val Glu Ala Ala Lys Gly Thr
1355 1360 1365
Asn Leu Phe Phe Ala Val Phe Glu Lys Asp Gly Lys Arg Ser Tyr
1370 1375 1380
Leu Thr Ile Pro Leu Asn Val Met Ile Asp Cys Gln Lys Lys Tyr
1385 1390 1395
Gly Asn Gln Trp Lys Gln Asn Ile Glu Ser Tyr Leu Lys Glu Lys
1400 1405 1410
Asp Leu Val Glu Lys Asp Val Gln Leu Leu Phe Ile Leu Ser Pro
1415 1420 1425
Asn Asp Leu Val Tyr Leu Pro Thr Glu Asn Glu Leu Lys Lys Gly
1430 1435 1440
Ile Thr Asn Pro Asp Lys Asp Gln Ile Tyr Lys Phe Val Ser Cys
1445 1450 1455
Thr Ser Asn Glu Ala His Phe Ile Pro Ser Phe Val Ala Asn Pro
1460 1465 1470
Ile Val Gln Thr Thr Glu Leu Gly Ser Asn Asn Lys Ala Gln Arg
1475 1480 1485
Ala Trp Asn Asn Lys Met Ile Lys Glu Ile Cys Ile Pro Ile Glu
1490 1495 1500
Val Asp Arg Leu Gly Asn Ile Lys
1505 1510
<210> 56
<211> 1511
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 56
Met Lys Lys Ile Leu Gly Leu Ala Leu Gly Thr Asn Ser Ile Gly Trp
1 5 10 15
Ala Val Val Asn Ala Asp Ala Ile Thr Arg Asn Asp Gly Ser Arg Tyr
20 25 30
Leu Lys Pro Asn Ser Ile Ser Ala Ala Gly Ser Arg Ile Ile Pro Met
35 40 45
Ser Ala Asp Val Leu Gly Asn Phe Glu Ser Gly Ile Thr Val Ser Gln
50 55 60
Thr Lys Asp Arg Thr Asp Lys Arg Met Ala Arg Arg Leu His Glu Arg
65 70 75 80
Ala Leu Leu Arg Arg Glu Arg Leu Leu Arg Ile Leu Ser Leu Met Asp
85 90 95
Phe Leu Pro Lys His Phe Ala Ser Lys Ile Asn Arg Tyr Gly Lys Phe
100 105 110
Thr Asp Asp Ser Glu Pro Lys Leu Ala Trp Arg Lys Asn Thr Glu Gly
115 120 125
Lys Tyr Glu Phe Ile Phe Gln Asp Ala Phe Asn Glu Met Leu Ala Glu
130 135 140
Phe Lys Asp Lys Gln Pro Glu Ile Val Lys Glu Gly Lys Lys Ile Pro
145 150 155 160
Tyr Asp Trp Thr Ile Tyr Tyr Leu Arg Lys Lys Ala Leu Glu Lys Ala
165 170 175
Leu Ser Lys Glu Glu Leu Ser Trp Leu Leu Leu Gln Phe Asn Gln Lys
180 185 190
Arg Gly Tyr Tyr Gln Leu Arg Gly Glu Glu Glu Asp Ile Pro Gln Asp
195 200 205
Lys Lys Ile Glu Tyr Leu Ala Gln Lys Val Val Lys Val Glu Ala Thr
210 215 220
Asp Gln Lys Lys Gly Asp Asp Ile Trp Tyr Asn Val Tyr Leu Glu Asn
225 230 235 240
Gly Met Ile Tyr Arg Arg Thr Ser Lys Ala Pro Leu Asp Trp Glu Gly
245 250 255
Lys Ile Lys Glu Phe Ile Val Thr Thr Asp Leu Glu Lys Asp Gly Thr
260 265 270
Pro Lys Lys Asp Lys Glu Gly Asn Ile Lys Arg Ser Phe Arg Ala Pro
275 280 285
Gln Glu Asp Asp Trp Thr Leu Leu Lys Lys Lys Thr Glu Ala Asp Ile
290 295 300
Glu Lys Ser Thr Lys Thr Val Gly Cys Tyr Ile Tyr Asp Ser Leu Leu
305 310 315 320
Asn Asn Pro Lys Gln Lys Ile Ile Gly Lys Leu Val Arg Thr Val Glu
325 330 335
Arg Lys Phe Tyr Lys Glu Glu Leu Thr Gln Ile Leu Lys Lys Gln Val
340 345 350
Glu Leu Ile Pro Glu Leu Arg Asn Asp Asn Leu Tyr Lys Gln Cys Ile
355 360 365
Glu Glu Leu Tyr Pro Ile Asn Glu Ala His Arg Asn Thr Ile Ala Lys
370 375 380
Thr Asp Phe Ala Asn Leu Phe Ile Asn Asp Ile Leu Phe Tyr Gln Arg
385 390 395 400
Pro Leu Lys Ser Lys Lys Ser Gln Ile Asp Asn Cys Pro Tyr Glu Glu
405 410 415
His Ile Phe Ile Asp Ser Lys Thr Gly Glu Lys Lys Lys Val Pro Val
420 425 430
Lys Cys Ile Thr Lys Ser Asn Pro Leu Phe Gln Glu Phe Arg Leu Trp
435 440 445
Gln Phe Ile Gln Asn Leu Arg Ile Tyr Gln Arg Glu Lys Glu Ile Asp
450 455 460
Gly Lys Leu Ser Thr Asp Val Asp Ile Thr Ser Glu Cys Leu Lys Ser
465 470 475 480
Glu Glu Asp Tyr Val Arg Leu Phe Asp Trp Leu Asn Asp Arg Glu Ser
485 490 495
Ile Glu Gln Glu Glu Leu Leu Lys Tyr Leu Phe Asn Thr Lys Lys Ser
500 505 510
Lys Asn Lys Glu Asn Pro Tyr Arg Trp Asn Tyr Val Glu Asp Lys Val
515 520 525
Tyr Pro Cys Asn Glu Thr Arg Ala Thr Ile Leu Lys Gly Leu Ser Lys
530 535 540
Cys Gly Ile Asn Ala Ser Val Leu Ser Ser Glu Met Glu Met Ala Leu
545 550 555 560
Trp His Ile Leu Tyr Ser Val Glu Asp Lys Lys Glu Ile Glu Thr Ala
565 570 575
Leu Thr His Phe Ala Gln Lys Gln Gly Trp Asn Gly Glu Phe Ala Ile
580 585 590
Val Phe Ser Lys Leu Lys Pro Phe Lys Lys Asp Tyr Gly Ser Tyr Ser
595 600 605
Glu Lys Ala Ile Lys Lys Leu Leu Ser Leu Met Arg Met Gly Lys Tyr
610 615 620
Trp Asn Gln Asp Asn Ile Asp Lys Asn Thr Leu Asp Arg Ile Asp Lys
625 630 635 640
Ile Ile Asn Gly Glu Tyr Asp Glu Lys Ile Ser Asn Arg Val Arg Asp
645 650 655
Asn Ala Ile Asn Leu Lys Asp Ile Ser Asp Phe Arg Gly Leu Pro Val
660 665 670
Trp Leu Ala Cys Tyr Ile Val Tyr Asp Arg His Ser Glu Ala Lys Asp
675 680 685
Cys Thr Lys Trp Asn Thr Pro Glu Glu Ile Asp Ser Tyr Leu Lys Lys
690 695 700
Phe Lys Gln His Ser Leu Arg Asn Pro Ile Val Glu Gln Val Val Thr
705 710 715 720
Glu Thr Leu Arg Thr Val Arg Asp Ile Trp Lys Gln Glu Gly Gln Ile
725 730 735
Asp Glu Ile His Leu Glu Leu Gly Arg Asp Leu Lys Asn Pro Ala Asp
740 745 750
Lys Arg Lys Lys Met Ser Glu Asn Ile Leu Lys Asn Glu Asn Thr Asn
755 760 765
Leu Arg Ile Lys Ala Met Leu Met Glu Phe Met Asn Pro Gly Met Gly
770 775 780
Ile Glu Asn Val Arg Pro Tyr Ser Pro Ser Gln Gln Asp Ile Leu Arg
785 790 795 800
Ile Tyr Glu Glu Asn Ala Leu Glu Asn Leu Thr Lys Asp Asp Glu Glu
805 810 815
Phe Asp Phe Ile Ser Lys Ile Ser Lys Gln Ala Gln Pro Thr Lys Ser
820 825 830
Asp Ile Val Arg Tyr Lys Cys Trp Leu Glu Gln Lys Tyr Arg Ser Pro
835 840 845
Tyr Thr Gly Lys Thr Ile Ser Leu Ser Lys Leu Phe Thr Ser Ala Tyr
850 855 860
Glu Ile Glu Ala Ile Ile Pro Gln Ser Arg Tyr Phe Asp Asp Ser Phe
865 870 875 880
Ser Asn Lys Val Ile Cys Glu Ala Glu Val Asn Lys Leu Lys Asp Arg
885 890 895
Gln Leu Gly His Glu Phe Ile Glu Glu His His Gly Glu Lys Val Gln
900 905 910
Leu Ser Gln Gly Glu Val Val Glu Ile Leu Ser Val Asp Ala Tyr Glu
915 920 925
Lys Phe Val Lys Glu Asn Tyr Ala Asn Asn Arg Val Lys Met Lys Lys
930 935 940
Leu Leu Met Glu Asn Ile Pro Asp Glu Phe Ile Glu Arg Gln Leu Asn
945 950 955 960
Asp Ser Arg Tyr Ile Ser Lys Val Val Lys Gly Leu Leu Ser Asn Ile
965 970 975
Val Arg Glu Lys Ile Asp Asp Glu Asn Tyr Glu Pro Glu Ala Val Ser
980 985 990
Lys Asn Leu Ile Ser Cys Asn Gly Ala Val Thr Asp Arg Leu Lys Lys
995 1000 1005
Asp Trp Gly Met Asn Asp Val Trp Asn Ser Ile Ile Leu Pro Arg
1010 1015 1020
Phe Ile Arg Met Asn Gln Ile Thr Gly Lys Asp Cys Phe Thr Thr
1025 1030 1035
Thr Asn Ala Glu Gly His Leu Ile Pro Gln Met Pro Leu Glu Leu
1040 1045 1050
Gln Lys Gly Phe Asn Lys Lys Arg Ile Asp His Arg His His Ala
1055 1060 1065
Met Asp Ala Ile Val Ile Ala Cys Thr Thr Arg Asp His Val Asn
1070 1075 1080
Leu Leu Asn Asn Glu Ala Ala His Ser Lys Phe Asn Ala Thr Arg
1085 1090 1095
Tyr Gln Leu Gln Arg Lys Leu Arg Cys Phe Glu Lys Ala Met Ile
1100 1105 1110
Asp Gly Lys Glu Arg Glu Val Ala Lys Glu Phe Leu Lys Pro Trp
1115 1120 1125
Asp Ser Phe Thr Met Asp Ser Lys Asn Ile Leu Glu Asn Ile Ile
1130 1135 1140
Val Ser Phe Lys Gln Asn Gln Arg Val Ile Asn Lys Thr Thr Asn
1145 1150 1155
Thr Phe Gln His Phe Asp Glu Asn Gly Lys Lys Thr Phe Val Lys
1160 1165 1170
Gln Gly Lys Gly Asn Ser Trp Ala Ile Arg Lys Pro Met His Lys
1175 1180 1185
Asp Thr Val Phe Gly Glu Ile Asn Leu Arg Lys Val Lys Ser Val
1190 1195 1200
Ser Leu Ser Asp Ala Ile Lys Val Pro Glu Arg Ile Leu Asn Lys
1205 1210 1215
Arg Ile Lys Glu Lys Ile Thr Glu Leu Lys Asn Asn Lys Val Asp
1220 1225 1230
Ala Lys Asn Ile Lys Lys Tyr Ile Glu Glu Tyr His Ile Gly Gly
1235 1240 1245
Tyr Gly Ile Asp Thr Ser Lys Ile Asp Val Phe Tyr Phe Thr Lys
1250 1255 1260
Glu Thr Lys Glu Arg Phe Phe Ala Thr Arg Lys Ser Leu Asp Ser
1265 1270 1275
Ser Phe Asn Gln Ala Lys Ile Glu Asp Ser Ile Ala Asp Ser Gly
1280 1285 1290
Ile Gln Lys Ile Leu Leu Ala His Leu Lys Ser Lys Asn Gly Asp
1295 1300 1305
Ala Glu Gln Ala Phe Ser Pro Asp Gly Ile Asp Glu Met Asn Lys
1310 1315 1320
Asn Ile Val Glu Leu Asn Asn Gly Lys Phe His Gln Pro Ile Leu
1325 1330 1335
Lys Val Arg Val Tyr Glu Lys Ala Asp Lys Phe Ala Val Gly Gln
1340 1345 1350
Lys Gly Asn Lys Lys Val Lys Phe Val Glu Ala Ala Lys Gly Thr
1355 1360 1365
Asn Leu Phe Phe Ala Val Phe Glu Lys Asp Gly Lys Arg Ser Tyr
1370 1375 1380
Leu Thr Ile Pro Leu Asn Val Met Ile Asp Cys Gln Lys Lys Tyr
1385 1390 1395
Gly Asn Gln Trp Lys Gln Asn Ile Glu Ser Tyr Leu Lys Glu Lys
1400 1405 1410
Asp Leu Val Glu Lys Asp Val Gln Leu Leu Phe Ile Leu Ser Pro
1415 1420 1425
Asn Asp Leu Val Tyr Leu Pro Thr Glu Asn Glu Leu Lys Lys Gly
1430 1435 1440
Ile Thr Asn Pro Asp Lys Asp Gln Ile Tyr Lys Phe Val Ser Cys
1445 1450 1455
Thr Ser Asn Glu Ala His Phe Ile Pro Ser Phe Val Ala Asn Pro
1460 1465 1470
Ile Val Gln Thr Thr Glu Leu Gly Ser Asn Asn Lys Ala Gln Arg
1475 1480 1485
Ala Trp Asn Asn Lys Met Ile Lys Glu Ile Cys Ile Pro Ile Glu
1490 1495 1500
Val Asp Arg Leu Gly Asn Ile Lys
1505 1510
<210> 57
<211> 1337
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 57
Met Lys Lys Asp Tyr Val Ile Gly Leu Ala Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Met Thr Glu Asp Tyr Gln Leu Val Lys Lys Lys Met
20 25 30
Pro Ile Tyr Gly Asn Thr Glu Lys Lys Lys Ile Lys Lys Asn Phe Trp
35 40 45
Gly Val Arg Leu Phe Glu Glu Gly His Thr Ala Glu Asp Arg Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Ile Ser Arg Arg Arg Asn Arg Leu Arg
65 70 75 80
Tyr Leu Gln Ala Phe Phe Glu Glu Ala Met Thr Ala Leu Asp Glu Asn
85 90 95
Phe Phe Ala Arg Leu Gln Glu Ser Phe Leu Val Pro Glu Asp Lys Lys
100 105 110
Trp His Arg His Pro Ile Phe Ala Lys Leu Glu Asp Glu Val Ala Tyr
115 120 125
His Glu Thr Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp
130 135 140
Ser Ser Glu Gln Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His
145 150 155 160
Ile Val Lys Tyr Arg Gly His Phe Leu Ile Glu Gly Lys Leu Ser Thr
165 170 175
Glu Asn Ile Ser Val Lys Glu Gln Phe Gln Gln Phe Met Ile Ile Tyr
180 185 190
Asn Gln Thr Phe Val Asn Gly Glu Ser Arg Leu Val Ser Ala Pro Leu
195 200 205
Pro Glu Ser Val Leu Ile Glu Glu Glu Leu Thr Glu Lys Ala Ser Arg
210 215 220
Thr Lys Lys Ser Glu Lys Val Leu Gln Gln Phe Pro Gln Glu Lys Ala
225 230 235 240
Asn Gly Leu Phe Gly Gln Phe Leu Lys Leu Met Val Gly Asn Lys Ala
245 250 255
Asp Phe Lys Lys Val Phe Gly Leu Glu Glu Glu Ala Lys Ile Thr Tyr
260 265 270
Ala Ser Glu Ser Tyr Glu Glu Asp Leu Glu Gly Ile Leu Ala Lys Val
275 280 285
Gly Asp Glu Tyr Ser Asp Val Phe Leu Ala Ala Lys Asn Val Tyr Asp
290 295 300
Ala Val Glu Leu Ser Thr Ile Leu Ala Asp Ser Asp Lys Lys Ser His
305 310 315 320
Ala Lys Leu Ser Ser Ser Met Ile Val Arg Phe Thr Glu His Gln Glu
325 330 335
Asp Leu Lys Asn Phe Lys Arg Phe Ile Arg Glu Asn Cys Pro Asp Glu
340 345 350
Tyr Asp Asn Leu Phe Lys Asn Glu Gln Lys Asp Gly Tyr Ala Gly Tyr
355 360 365
Ile Ala His Ala Gly Lys Val Ser Gln Leu Lys Phe Tyr Gln Tyr Val
370 375 380
Lys Lys Ile Ile Gln Asp Ile Ala Gly Ala Glu Tyr Phe Leu Glu Lys
385 390 395 400
Ile Ala Gln Glu Asn Phe Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly
405 410 415
Val Ile Pro His Gln Ile His Leu Ala Glu Leu Gln Ala Ile Ile His
420 425 430
Arg Gln Ala Ala Tyr Tyr Pro Phe Leu Lys Glu Asn Gln Lys Lys Ile
435 440 445
Glu Gln Leu Val Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ser
450 455 460
Lys Gly Asp Ala Ser Thr Phe Ala Trp Leu Lys Arg Gln Ser Glu Glu
465 470 475 480
Pro Ile Arg Pro Trp Asn Leu Gln Glu Thr Val Asp Leu Asp Gln Ser
485 490 495
Ala Thr Ala Phe Ile Glu Arg Met Thr Asn Phe Asp Thr Tyr Leu Pro
500 505 510
Ser Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Lys Phe Met
515 520 525
Val Phe Asn Glu Leu Thr Lys Ile Ser Tyr Thr Asp Asp Arg Gly Ile
530 535 540
Lys Ala Asn Phe Ser Gly Lys Glu Lys Glu Lys Ile Phe Asp Tyr Leu
545 550 555 560
Phe Lys Thr Arg Arg Lys Val Lys Lys Lys Asp Ile Ile Gln Phe Tyr
565 570 575
Arg Asn Glu Tyr Asn Thr Glu Ile Val Thr Leu Ser Gly Leu Glu Glu
580 585 590
Asp Gln Phe Asn Ala Ser Phe Ser Thr Tyr Gln Asp Leu Leu Lys Cys
595 600 605
Gly Leu Thr Arg Ala Glu Leu Asp His Pro Asp Asn Ala Glu Lys Leu
610 615 620
Glu Asp Ile Ile Lys Ile Leu Thr Ile Phe Glu Asp Arg Gln Arg Ile
625 630 635 640
Arg Thr Gln Leu Ser Thr Phe Lys Gly Gln Phe Ser Glu Glu Val Leu
645 650 655
Lys Lys Leu Glu Arg Lys His Tyr Thr Gly Trp Gly Arg Leu Ser Lys
660 665 670
Lys Leu Ile Asn Gly Ile Tyr Asp Lys Glu Ser Gly Lys Thr Ile Leu
675 680 685
Asp Tyr Leu Ile Lys Asp Asp Gly Val Ser Lys His Tyr Asn Arg Asn
690 695 700
Phe Met Gln Leu Ile Asn Asp Ser Gln Leu Ser Phe Lys Asn Ala Ile
705 710 715 720
Gln Lys Ala Gln Ser Ser Glu His Glu Glu Thr Leu Ser Glu Thr Val
725 730 735
Asn Glu Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Tyr Gln Ser
740 745 750
Leu Lys Ile Val Asp Glu Leu Val Ala Ile Met Gly Tyr Ala Pro Lys
755 760 765
Arg Ile Val Val Glu Met Ala Arg Glu Asn Gln Thr Thr Ser Thr Gly
770 775 780
Lys Arg Arg Ser Ile Gln Arg Leu Lys Ile Val Glu Lys Ala Met Ala
785 790 795 800
Glu Ile Gly Ser Asn Leu Leu Lys Glu Gln Pro Thr Thr Asn Glu Gln
805 810 815
Leu Arg Asp Thr Arg Leu Phe Leu Tyr Tyr Met Gln Asn Gly Lys Asp
820 825 830
Met Tyr Thr Gly Asp Glu Leu Ser Leu His Arg Leu Ser His Tyr Asp
835 840 845
Ile Asp His Ile Ile Pro Gln Ser Phe Met Lys Asp Asp Ser Leu Asp
850 855 860
Asn Leu Val Leu Val Gly Ser Thr Glu Asn Arg Gly Lys Ser Asp Asp
865 870 875 880
Val Pro Ser Lys Glu Val Val Lys Lys Met Lys Ala Tyr Trp Glu Lys
885 890 895
Leu Tyr Ala Ala Gly Leu Ile Ser Gln Arg Lys Phe Gln Arg Leu Thr
900 905 910
Lys Gly Glu Gln Gly Gly Leu Thr Leu Glu Asp Lys Ala His Phe Ile
915 920 925
Gln Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys Asn Val Ala Gly
930 935 940
Ile Leu Asp Gln Arg Tyr Asn Ala Lys Ser Lys Glu Lys Lys Val Gln
945 950 955 960
Ile Ile Thr Leu Lys Ala Ser Leu Thr Ser Gln Phe Arg Ser Ile Phe
965 970 975
Gly Leu Tyr Lys Val Arg Glu Val Asn Asp Tyr His His Gly Gln Asp
980 985 990
Ala Tyr Leu Asn Cys Val Val Ala Thr Thr Leu Leu Lys Val Tyr Pro
995 1000 1005
Asn Leu Ala Pro Glu Phe Val Tyr Gly Glu Tyr Pro Lys Phe Gln
1010 1015 1020
Ala Phe Lys Glu Asn Lys Ala Thr Ala Lys Ala Ile Ile Tyr Thr
1025 1030 1035
Asn Leu Leu Arg Phe Phe Thr Glu Asp Glu Pro Arg Phe Thr Lys
1040 1045 1050
Asp Gly Glu Ile Leu Trp Ser Asn Ser Tyr Leu Lys Thr Ile Lys
1055 1060 1065
Lys Glu Leu Asn Tyr His Gln Met Asn Ile Val Lys Lys Val Glu
1070 1075 1080
Val Gln Lys Gly Gly Phe Ser Lys Glu Ser Ile Lys Pro Lys Gly
1085 1090 1095
Pro Ser Asn Lys Leu Ile Pro Val Lys Asn Gly Leu Asp Pro Gln
1100 1105 1110
Lys Tyr Gly Gly Phe Asp Ser Pro Val Val Ala Tyr Thr Val Leu
1115 1120 1125
Phe Thr His Glu Lys Gly Lys Lys Pro Leu Ile Lys Gln Glu Ile
1130 1135 1140
Leu Gly Ile Thr Ile Met Glu Lys Thr Arg Phe Glu Gln Asn Pro
1145 1150 1155
Ile Leu Phe Leu Glu Glu Lys Gly Phe Leu Arg Pro Arg Val Leu
1160 1165 1170
Met Lys Leu Pro Lys Tyr Thr Leu Tyr Glu Phe Pro Glu Gly Arg
1175 1180 1185
Arg Arg Leu Leu Ala Ser Ala Lys Glu Ala Gln Lys Gly Asn Gln
1190 1195 1200
Met Val Leu Pro Glu His Leu Leu Thr Leu Leu Tyr His Ala Lys
1205 1210 1215
Gln Cys Leu Leu Pro Asn Gln Ser Glu Ser Leu Ala Tyr Val Glu
1220 1225 1230
Gln His Gln Pro Glu Phe Gln Glu Ile Leu Glu Arg Val Val Asp
1235 1240 1245
Phe Ala Glu Val His Thr Leu Ala Lys Ser Lys Val Gln Gln Ile
1250 1255 1260
Val Lys Leu Phe Glu Ala Asn Gln Thr Ala Asp Val Lys Glu Ile
1265 1270 1275
Ala Ala Ser Phe Ile Gln Leu Met Gln Phe Asn Ala Met Gly Ala
1280 1285 1290
Pro Ser Thr Phe Lys Phe Phe Gln Lys Asp Ile Glu Arg Ala Arg
1295 1300 1305
Tyr Thr Ser Ile Lys Glu Ile Phe Asp Ala Thr Ile Ile Tyr Gln
1310 1315 1320
Ser Thr Thr Gly Leu Tyr Glu Thr Arg Arg Lys Val Val Asp
1325 1330 1335
<210> 58
<211> 1337
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 58
Met Lys Lys Asp Tyr Val Ile Gly Leu Asp Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Met Thr Glu Asp Tyr Gln Leu Val Lys Lys Lys Met
20 25 30
Pro Ile Tyr Gly Asn Thr Glu Lys Lys Lys Ile Lys Lys Asn Phe Trp
35 40 45
Gly Val Arg Leu Phe Glu Glu Gly His Thr Ala Glu Asp Arg Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Ile Ser Arg Arg Arg Asn Arg Leu Arg
65 70 75 80
Tyr Leu Gln Ala Phe Phe Glu Glu Ala Met Thr Ala Leu Asp Glu Asn
85 90 95
Phe Phe Ala Arg Leu Gln Glu Ser Phe Leu Val Pro Glu Asp Lys Lys
100 105 110
Trp His Arg His Pro Ile Phe Ala Lys Leu Glu Asp Glu Val Ala Tyr
115 120 125
His Glu Thr Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp
130 135 140
Ser Ser Glu Gln Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His
145 150 155 160
Ile Val Lys Tyr Arg Gly His Phe Leu Ile Glu Gly Lys Leu Ser Thr
165 170 175
Glu Asn Ile Ser Val Lys Glu Gln Phe Gln Gln Phe Met Ile Ile Tyr
180 185 190
Asn Gln Thr Phe Val Asn Gly Glu Ser Arg Leu Val Ser Ala Pro Leu
195 200 205
Pro Glu Ser Val Leu Ile Glu Glu Glu Leu Thr Glu Lys Ala Ser Arg
210 215 220
Thr Lys Lys Ser Glu Lys Val Leu Gln Gln Phe Pro Gln Glu Lys Ala
225 230 235 240
Asn Gly Leu Phe Gly Gln Phe Leu Lys Leu Met Val Gly Asn Lys Ala
245 250 255
Asp Phe Lys Lys Val Phe Gly Leu Glu Glu Glu Ala Lys Ile Thr Tyr
260 265 270
Ala Ser Glu Ser Tyr Glu Glu Asp Leu Glu Gly Ile Leu Ala Lys Val
275 280 285
Gly Asp Glu Tyr Ser Asp Val Phe Leu Ala Ala Lys Asn Val Tyr Asp
290 295 300
Ala Val Glu Leu Ser Thr Ile Leu Ala Asp Ser Asp Lys Lys Ser His
305 310 315 320
Ala Lys Leu Ser Ser Ser Met Ile Val Arg Phe Thr Glu His Gln Glu
325 330 335
Asp Leu Lys Asn Phe Lys Arg Phe Ile Arg Glu Asn Cys Pro Asp Glu
340 345 350
Tyr Asp Asn Leu Phe Lys Asn Glu Gln Lys Asp Gly Tyr Ala Gly Tyr
355 360 365
Ile Ala His Ala Gly Lys Val Ser Gln Leu Lys Phe Tyr Gln Tyr Val
370 375 380
Lys Lys Ile Ile Gln Asp Ile Ala Gly Ala Glu Tyr Phe Leu Glu Lys
385 390 395 400
Ile Ala Gln Glu Asn Phe Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly
405 410 415
Val Ile Pro His Gln Ile His Leu Ala Glu Leu Gln Ala Ile Ile His
420 425 430
Arg Gln Ala Ala Tyr Tyr Pro Phe Leu Lys Glu Asn Gln Lys Lys Ile
435 440 445
Glu Gln Leu Val Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ser
450 455 460
Lys Gly Asp Ala Ser Thr Phe Ala Trp Leu Lys Arg Gln Ser Glu Glu
465 470 475 480
Pro Ile Arg Pro Trp Asn Leu Gln Glu Thr Val Asp Leu Asp Gln Ser
485 490 495
Ala Thr Ala Phe Ile Glu Arg Met Thr Asn Phe Asp Thr Tyr Leu Pro
500 505 510
Ser Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Lys Phe Met
515 520 525
Val Phe Asn Glu Leu Thr Lys Ile Ser Tyr Thr Asp Asp Arg Gly Ile
530 535 540
Lys Ala Asn Phe Ser Gly Lys Glu Lys Glu Lys Ile Phe Asp Tyr Leu
545 550 555 560
Phe Lys Thr Arg Arg Lys Val Lys Lys Lys Asp Ile Ile Gln Phe Tyr
565 570 575
Arg Asn Glu Tyr Asn Thr Glu Ile Val Thr Leu Ser Gly Leu Glu Glu
580 585 590
Asp Gln Phe Asn Ala Ser Phe Ser Thr Tyr Gln Asp Leu Leu Lys Cys
595 600 605
Gly Leu Thr Arg Ala Glu Leu Asp His Pro Asp Asn Ala Glu Lys Leu
610 615 620
Glu Asp Ile Ile Lys Ile Leu Thr Ile Phe Glu Asp Arg Gln Arg Ile
625 630 635 640
Arg Thr Gln Leu Ser Thr Phe Lys Gly Gln Phe Ser Glu Glu Val Leu
645 650 655
Lys Lys Leu Glu Arg Lys His Tyr Thr Gly Trp Gly Arg Leu Ser Lys
660 665 670
Lys Leu Ile Asn Gly Ile Tyr Asp Lys Glu Ser Gly Lys Thr Ile Leu
675 680 685
Asp Tyr Leu Ile Lys Asp Asp Gly Val Ser Lys His Tyr Asn Arg Asn
690 695 700
Phe Met Gln Leu Ile Asn Asp Ser Gln Leu Ser Phe Lys Asn Ala Ile
705 710 715 720
Gln Lys Ala Gln Ser Ser Glu His Glu Glu Thr Leu Ser Glu Thr Val
725 730 735
Asn Glu Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Tyr Gln Ser
740 745 750
Leu Lys Ile Val Asp Glu Leu Val Ala Ile Met Gly Tyr Ala Pro Lys
755 760 765
Arg Ile Val Val Glu Met Ala Arg Glu Asn Gln Thr Thr Ser Thr Gly
770 775 780
Lys Arg Arg Ser Ile Gln Arg Leu Lys Ile Val Glu Lys Ala Met Ala
785 790 795 800
Glu Ile Gly Ser Asn Leu Leu Lys Glu Gln Pro Thr Thr Asn Glu Gln
805 810 815
Leu Arg Asp Thr Arg Leu Phe Leu Tyr Tyr Met Gln Asn Gly Lys Asp
820 825 830
Met Tyr Thr Gly Asp Glu Leu Ser Leu His Arg Leu Ser Ala Tyr Asp
835 840 845
Ile Asp His Ile Ile Pro Gln Ser Phe Met Lys Asp Asp Ser Leu Asp
850 855 860
Asn Leu Val Leu Val Gly Ser Thr Glu Asn Arg Gly Lys Ser Asp Asp
865 870 875 880
Val Pro Ser Lys Glu Val Val Lys Lys Met Lys Ala Tyr Trp Glu Lys
885 890 895
Leu Tyr Ala Ala Gly Leu Ile Ser Gln Arg Lys Phe Gln Arg Leu Thr
900 905 910
Lys Gly Glu Gln Gly Gly Leu Thr Leu Glu Asp Lys Ala His Phe Ile
915 920 925
Gln Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys Asn Val Ala Gly
930 935 940
Ile Leu Asp Gln Arg Tyr Asn Ala Lys Ser Lys Glu Lys Lys Val Gln
945 950 955 960
Ile Ile Thr Leu Lys Ala Ser Leu Thr Ser Gln Phe Arg Ser Ile Phe
965 970 975
Gly Leu Tyr Lys Val Arg Glu Val Asn Asp Tyr His His Gly Gln Asp
980 985 990
Ala Tyr Leu Asn Cys Val Val Ala Thr Thr Leu Leu Lys Val Tyr Pro
995 1000 1005
Asn Leu Ala Pro Glu Phe Val Tyr Gly Glu Tyr Pro Lys Phe Gln
1010 1015 1020
Ala Phe Lys Glu Asn Lys Ala Thr Ala Lys Ala Ile Ile Tyr Thr
1025 1030 1035
Asn Leu Leu Arg Phe Phe Thr Glu Asp Glu Pro Arg Phe Thr Lys
1040 1045 1050
Asp Gly Glu Ile Leu Trp Ser Asn Ser Tyr Leu Lys Thr Ile Lys
1055 1060 1065
Lys Glu Leu Asn Tyr His Gln Met Asn Ile Val Lys Lys Val Glu
1070 1075 1080
Val Gln Lys Gly Gly Phe Ser Lys Glu Ser Ile Lys Pro Lys Gly
1085 1090 1095
Pro Ser Asn Lys Leu Ile Pro Val Lys Asn Gly Leu Asp Pro Gln
1100 1105 1110
Lys Tyr Gly Gly Phe Asp Ser Pro Val Val Ala Tyr Thr Val Leu
1115 1120 1125
Phe Thr His Glu Lys Gly Lys Lys Pro Leu Ile Lys Gln Glu Ile
1130 1135 1140
Leu Gly Ile Thr Ile Met Glu Lys Thr Arg Phe Glu Gln Asn Pro
1145 1150 1155
Ile Leu Phe Leu Glu Glu Lys Gly Phe Leu Arg Pro Arg Val Leu
1160 1165 1170
Met Lys Leu Pro Lys Tyr Thr Leu Tyr Glu Phe Pro Glu Gly Arg
1175 1180 1185
Arg Arg Leu Leu Ala Ser Ala Lys Glu Ala Gln Lys Gly Asn Gln
1190 1195 1200
Met Val Leu Pro Glu His Leu Leu Thr Leu Leu Tyr His Ala Lys
1205 1210 1215
Gln Cys Leu Leu Pro Asn Gln Ser Glu Ser Leu Ala Tyr Val Glu
1220 1225 1230
Gln His Gln Pro Glu Phe Gln Glu Ile Leu Glu Arg Val Val Asp
1235 1240 1245
Phe Ala Glu Val His Thr Leu Ala Lys Ser Lys Val Gln Gln Ile
1250 1255 1260
Val Lys Leu Phe Glu Ala Asn Gln Thr Ala Asp Val Lys Glu Ile
1265 1270 1275
Ala Ala Ser Phe Ile Gln Leu Met Gln Phe Asn Ala Met Gly Ala
1280 1285 1290
Pro Ser Thr Phe Lys Phe Phe Gln Lys Asp Ile Glu Arg Ala Arg
1295 1300 1305
Tyr Thr Ser Ile Lys Glu Ile Phe Asp Ala Thr Ile Ile Tyr Gln
1310 1315 1320
Ser Thr Thr Gly Leu Tyr Glu Thr Arg Arg Lys Val Val Asp
1325 1330 1335
<210> 59
<211> 1337
<212> PRT
<213> 人工序列(Artificial Sequence)
<220>
<223> 人工突变的序列
<400> 59
Met Lys Lys Asp Tyr Val Ile Gly Leu Ala Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Met Thr Glu Asp Tyr Gln Leu Val Lys Lys Lys Met
20 25 30
Pro Ile Tyr Gly Asn Thr Glu Lys Lys Lys Ile Lys Lys Asn Phe Trp
35 40 45
Gly Val Arg Leu Phe Glu Glu Gly His Thr Ala Glu Asp Arg Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Ile Ser Arg Arg Arg Asn Arg Leu Arg
65 70 75 80
Tyr Leu Gln Ala Phe Phe Glu Glu Ala Met Thr Ala Leu Asp Glu Asn
85 90 95
Phe Phe Ala Arg Leu Gln Glu Ser Phe Leu Val Pro Glu Asp Lys Lys
100 105 110
Trp His Arg His Pro Ile Phe Ala Lys Leu Glu Asp Glu Val Ala Tyr
115 120 125
His Glu Thr Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp
130 135 140
Ser Ser Glu Gln Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His
145 150 155 160
Ile Val Lys Tyr Arg Gly His Phe Leu Ile Glu Gly Lys Leu Ser Thr
165 170 175
Glu Asn Ile Ser Val Lys Glu Gln Phe Gln Gln Phe Met Ile Ile Tyr
180 185 190
Asn Gln Thr Phe Val Asn Gly Glu Ser Arg Leu Val Ser Ala Pro Leu
195 200 205
Pro Glu Ser Val Leu Ile Glu Glu Glu Leu Thr Glu Lys Ala Ser Arg
210 215 220
Thr Lys Lys Ser Glu Lys Val Leu Gln Gln Phe Pro Gln Glu Lys Ala
225 230 235 240
Asn Gly Leu Phe Gly Gln Phe Leu Lys Leu Met Val Gly Asn Lys Ala
245 250 255
Asp Phe Lys Lys Val Phe Gly Leu Glu Glu Glu Ala Lys Ile Thr Tyr
260 265 270
Ala Ser Glu Ser Tyr Glu Glu Asp Leu Glu Gly Ile Leu Ala Lys Val
275 280 285
Gly Asp Glu Tyr Ser Asp Val Phe Leu Ala Ala Lys Asn Val Tyr Asp
290 295 300
Ala Val Glu Leu Ser Thr Ile Leu Ala Asp Ser Asp Lys Lys Ser His
305 310 315 320
Ala Lys Leu Ser Ser Ser Met Ile Val Arg Phe Thr Glu His Gln Glu
325 330 335
Asp Leu Lys Asn Phe Lys Arg Phe Ile Arg Glu Asn Cys Pro Asp Glu
340 345 350
Tyr Asp Asn Leu Phe Lys Asn Glu Gln Lys Asp Gly Tyr Ala Gly Tyr
355 360 365
Ile Ala His Ala Gly Lys Val Ser Gln Leu Lys Phe Tyr Gln Tyr Val
370 375 380
Lys Lys Ile Ile Gln Asp Ile Ala Gly Ala Glu Tyr Phe Leu Glu Lys
385 390 395 400
Ile Ala Gln Glu Asn Phe Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly
405 410 415
Val Ile Pro His Gln Ile His Leu Ala Glu Leu Gln Ala Ile Ile His
420 425 430
Arg Gln Ala Ala Tyr Tyr Pro Phe Leu Lys Glu Asn Gln Lys Lys Ile
435 440 445
Glu Gln Leu Val Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ser
450 455 460
Lys Gly Asp Ala Ser Thr Phe Ala Trp Leu Lys Arg Gln Ser Glu Glu
465 470 475 480
Pro Ile Arg Pro Trp Asn Leu Gln Glu Thr Val Asp Leu Asp Gln Ser
485 490 495
Ala Thr Ala Phe Ile Glu Arg Met Thr Asn Phe Asp Thr Tyr Leu Pro
500 505 510
Ser Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Lys Phe Met
515 520 525
Val Phe Asn Glu Leu Thr Lys Ile Ser Tyr Thr Asp Asp Arg Gly Ile
530 535 540
Lys Ala Asn Phe Ser Gly Lys Glu Lys Glu Lys Ile Phe Asp Tyr Leu
545 550 555 560
Phe Lys Thr Arg Arg Lys Val Lys Lys Lys Asp Ile Ile Gln Phe Tyr
565 570 575
Arg Asn Glu Tyr Asn Thr Glu Ile Val Thr Leu Ser Gly Leu Glu Glu
580 585 590
Asp Gln Phe Asn Ala Ser Phe Ser Thr Tyr Gln Asp Leu Leu Lys Cys
595 600 605
Gly Leu Thr Arg Ala Glu Leu Asp His Pro Asp Asn Ala Glu Lys Leu
610 615 620
Glu Asp Ile Ile Lys Ile Leu Thr Ile Phe Glu Asp Arg Gln Arg Ile
625 630 635 640
Arg Thr Gln Leu Ser Thr Phe Lys Gly Gln Phe Ser Glu Glu Val Leu
645 650 655
Lys Lys Leu Glu Arg Lys His Tyr Thr Gly Trp Gly Arg Leu Ser Lys
660 665 670
Lys Leu Ile Asn Gly Ile Tyr Asp Lys Glu Ser Gly Lys Thr Ile Leu
675 680 685
Asp Tyr Leu Ile Lys Asp Asp Gly Val Ser Lys His Tyr Asn Arg Asn
690 695 700
Phe Met Gln Leu Ile Asn Asp Ser Gln Leu Ser Phe Lys Asn Ala Ile
705 710 715 720
Gln Lys Ala Gln Ser Ser Glu His Glu Glu Thr Leu Ser Glu Thr Val
725 730 735
Asn Glu Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Tyr Gln Ser
740 745 750
Leu Lys Ile Val Asp Glu Leu Val Ala Ile Met Gly Tyr Ala Pro Lys
755 760 765
Arg Ile Val Val Glu Met Ala Arg Glu Asn Gln Thr Thr Ser Thr Gly
770 775 780
Lys Arg Arg Ser Ile Gln Arg Leu Lys Ile Val Glu Lys Ala Met Ala
785 790 795 800
Glu Ile Gly Ser Asn Leu Leu Lys Glu Gln Pro Thr Thr Asn Glu Gln
805 810 815
Leu Arg Asp Thr Arg Leu Phe Leu Tyr Tyr Met Gln Asn Gly Lys Asp
820 825 830
Met Tyr Thr Gly Asp Glu Leu Ser Leu His Arg Leu Ser Ala Tyr Asp
835 840 845
Ile Asp His Ile Ile Pro Gln Ser Phe Met Lys Asp Asp Ser Leu Asp
850 855 860
Asn Leu Val Leu Val Gly Ser Thr Glu Asn Arg Gly Lys Ser Asp Asp
865 870 875 880
Val Pro Ser Lys Glu Val Val Lys Lys Met Lys Ala Tyr Trp Glu Lys
885 890 895
Leu Tyr Ala Ala Gly Leu Ile Ser Gln Arg Lys Phe Gln Arg Leu Thr
900 905 910
Lys Gly Glu Gln Gly Gly Leu Thr Leu Glu Asp Lys Ala His Phe Ile
915 920 925
Gln Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys Asn Val Ala Gly
930 935 940
Ile Leu Asp Gln Arg Tyr Asn Ala Lys Ser Lys Glu Lys Lys Val Gln
945 950 955 960
Ile Ile Thr Leu Lys Ala Ser Leu Thr Ser Gln Phe Arg Ser Ile Phe
965 970 975
Gly Leu Tyr Lys Val Arg Glu Val Asn Asp Tyr His His Gly Gln Asp
980 985 990
Ala Tyr Leu Asn Cys Val Val Ala Thr Thr Leu Leu Lys Val Tyr Pro
995 1000 1005
Asn Leu Ala Pro Glu Phe Val Tyr Gly Glu Tyr Pro Lys Phe Gln
1010 1015 1020
Ala Phe Lys Glu Asn Lys Ala Thr Ala Lys Ala Ile Ile Tyr Thr
1025 1030 1035
Asn Leu Leu Arg Phe Phe Thr Glu Asp Glu Pro Arg Phe Thr Lys
1040 1045 1050
Asp Gly Glu Ile Leu Trp Ser Asn Ser Tyr Leu Lys Thr Ile Lys
1055 1060 1065
Lys Glu Leu Asn Tyr His Gln Met Asn Ile Val Lys Lys Val Glu
1070 1075 1080
Val Gln Lys Gly Gly Phe Ser Lys Glu Ser Ile Lys Pro Lys Gly
1085 1090 1095
Pro Ser Asn Lys Leu Ile Pro Val Lys Asn Gly Leu Asp Pro Gln
1100 1105 1110
Lys Tyr Gly Gly Phe Asp Ser Pro Val Val Ala Tyr Thr Val Leu
1115 1120 1125
Phe Thr His Glu Lys Gly Lys Lys Pro Leu Ile Lys Gln Glu Ile
1130 1135 1140
Leu Gly Ile Thr Ile Met Glu Lys Thr Arg Phe Glu Gln Asn Pro
1145 1150 1155
Ile Leu Phe Leu Glu Glu Lys Gly Phe Leu Arg Pro Arg Val Leu
1160 1165 1170
Met Lys Leu Pro Lys Tyr Thr Leu Tyr Glu Phe Pro Glu Gly Arg
1175 1180 1185
Arg Arg Leu Leu Ala Ser Ala Lys Glu Ala Gln Lys Gly Asn Gln
1190 1195 1200
Met Val Leu Pro Glu His Leu Leu Thr Leu Leu Tyr His Ala Lys
1205 1210 1215
Gln Cys Leu Leu Pro Asn Gln Ser Glu Ser Leu Ala Tyr Val Glu
1220 1225 1230
Gln His Gln Pro Glu Phe Gln Glu Ile Leu Glu Arg Val Val Asp
1235 1240 1245
Phe Ala Glu Val His Thr Leu Ala Lys Ser Lys Val Gln Gln Ile
1250 1255 1260
Val Lys Leu Phe Glu Ala Asn Gln Thr Ala Asp Val Lys Glu Ile
1265 1270 1275
Ala Ala Ser Phe Ile Gln Leu Met Gln Phe Asn Ala Met Gly Ala
1280 1285 1290
Pro Ser Thr Phe Lys Phe Phe Gln Lys Asp Ile Glu Arg Ala Arg
1295 1300 1305
Tyr Thr Ser Ile Lys Glu Ile Phe Asp Ala Thr Ile Ile Tyr Gln
1310 1315 1320
Ser Thr Thr Gly Leu Tyr Glu Thr Arg Arg Lys Val Val Asp
1325 1330 1335
<210> 60
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 60
gtagcggctg aagcactgca 20
<210> 61
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 61
cgccgtaggt 10
<210> 62
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 62
ggcgtgcagt gcttcagccg cta 23
<210> 63
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 63
ccccgaccac 10
<210> 64
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 64
gccgctaccc cgaccacatg aag 23
<210> 65
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 65
cagcacgact 10
<210> 66
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 66
ggccagggca cgggcagttt gcc 23
<210> 67
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 67
ggtggtgcag 10
<210> 68
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 68
gtggtgccca tcctggtcga gct 23
<210> 69
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 69
ggacggcgac 10
<210> 70
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 70
ggcatcgccc tcgccctcgc cgg 23
<210> 71
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 71
acacgctgaa 10
<210> 72
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 72
gccctcgccg gacacgctga act 23
<210> 73
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 73
tgtggccgtt 10
<210> 74
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 74
ggtggtcacc aaagtgggcc agg 23
<210> 75
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 75
gcacgggcag 10
<210> 76
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 76
gtgtccggcg agggcgaggg cga 23
<210> 77
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 77
tgccacctac 10
<210> 78
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 78
gtgagcaagg gcgaggagct gtt 23
<210> 79
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 79
caccggggtg 10
<210> 80
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 80
gtggtcacca aagtgggcca ggg 23
<210> 81
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 81
cacgggcagt 10
<210> 82
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 82
gcgagggcga gggcgatgcc acc 23
<210> 83
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 83
tacggcaagc 10
<210> 84
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 84
gtgctgcttc atgtggtcgg ggt 23
<210> 85
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 85
agcggctgaa 10
<210> 86
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 86
gctgaacttg tggccgttta cgt 23
<210> 87
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 87
cgccgtccag 10
<210> 88
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 88
gctgaagcac tgcacgccgt agg 23
<210> 89
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 89
tcagggtggt 10
<210> 90
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 90
gtcaccaaag tgggccaggg ca 22
<210> 91
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 91
cgggcagttt 10
<210> 92
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 92
gtaaacggcc acaagttcag cgt 23
<210> 93
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 93
gtccggcgag 10
<210> 94
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 94
gtggcatcgc cctcgccctc gcc 23
<210> 95
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 95
ggacacgctg 10
<210> 96
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 96
ggggtagcgg ctgaagcact gca 23
<210> 97
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 97
cgccgtaggt 10
<210> 98
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 98
ggcacgggca gtttgccggt ggt 23
<210> 99
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 99
gcagatgaac 10
<210> 100
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 100
ggggtggtgc ccatcctggt cga 23
<210> 101
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 101
gctggacggc 10
<210> 102
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 102
gcgtgcagtg cttcagccgc tac 23
<210> 103
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 103
cccgaccaca 10
<210> 104
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 104
gtcaccaaag tgggccaggg cac 23
<210> 105
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 105
gggcagtttg 10
<210> 106
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 106
gacggcgacg taaacggcca caa 23
<210> 107
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 107
gttcagcgtg 10
<210> 108
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 108
gcgtgcagtg cttcagccgc ta 22
<210> 109
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 109
ccccgaccac 10
<210> 110
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 110
gccacaagtt cagcgtgtcc ggc 23
<210> 111
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 111
gagggcgagg 10
<210> 112
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 112
gggtggtgcc catcctggtc gag 23
<210> 113
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 113
ctggacggcg 10
<210> 114
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 114
gtcagggtgg tcaccaaagt ggg 23
<210> 115
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 115
ccagggcacg 10
<210> 116
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 116
gtgcccatcc tggtcgagct gga 23
<210> 117
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 117
cggcgacgta 10
<210> 118
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 118
gccgtaggtg gcatcgccct cgc 23
<210> 119
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 119
cctcgccgga 10
<210> 120
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 120
ggcgacgtaa acggccacaa gtt 23
<210> 121
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 121
cagcgtgtcc 10
<210> 122
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 122
gagctggacg gcgacgtaaa cgg 23
<210> 123
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 123
ccacaagttc 10
<210> 124
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 124
gtggtgccca tcctggtcga 20
<210> 125
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 125
gctggacggc 10
<210> 126
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 126
ggccacaagt tcagcgtgtc cgg 23
<210> 127
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 127
cgagggcgag 10
<210> 128
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 128
ggtggtcacc aaagtgggcc agg 23
<210> 129
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 129
gcacgggcag 10
<210> 130
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 130
gtccggcgag ggcgagggcg atg 23
<210> 131
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 131
ccacctacgg 10
<210> 132
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 132
gtgcccatcc tggtcgagct gga 23
<210> 133
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 133
cggcgacgta 10
<210> 134
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 134
gtgtccggcg agggcgaggg cga 23
<210> 135
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 135
tgccacctac 10
<210> 136
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 136
gggtcagctt gccgtaggtg gca 23
<210> 137
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 137
tcgccctcgc 10
<210> 138
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 间隔区
<400> 138
gccgctaccc cgaccacatg aag 23
<210> 139
<211> 10
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> PAM
<400> 139
cagcacgact 10
<210> 140
<211> 3633
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 人源化的MAD 2007 V1
<400> 140
atggaaaatt ataggcaaaa gcacagattc gtcctcgcca cagatctggg aatcggaagc 60
aacggctggg ctatcatcga cctcgacgcc catagagtcg aggatctcgg cgtgcagatc 120
tttgagtccg gagaggaggg agccaagaaa gccagcgcta gagccagcca gcaaagaagg 180
ctcaagagga gcgcccacag actgaataga aggaagaaac aaaggaagga agccctcatc 240
aaattcctcc aagagatcga gtttcccgat ctcgtcgaga ttctcaactc cttcaagaag 300
cagaaaaacc ctaacgacat tctgtctctg agagtcaagg gactcgacaa caagctgtcc 360
cctctggagc tgttcagcat tctgatctac atgagcaaca acagaggata caaagacttc 420
tacgacaatg acattaacga caataatacc gacaaagacg aaaaggagat ggaaaaggcc 480
aagtccacca ttgagaagct cttcgcttcc aatagctaca gaaccgtggg agagatgatt 540
gccacagatc ctacctttat tgtggacaaa tccggcagca agaaagtcat caagtatcat 600
aacaaaaagg gataccagta tctgattcct agaaagctgc tggagaacga gatgtccctc 660
attctgcaca agcaagagga gttctacgat tgcctcagca tcgacaacat cacaattatt 720
ctggacaaga tttttttcca gagaaacttt gaggatggac ccggccccaa gaacaagagg 780
gatgattata aaaataattc caaaggaaat cagttttata ccggattcaa cgagatgatt 840
ggactgtgcc ccttctatcc taacgagaaa aagggcacca agaactctct gatctacgac 900
gagtactacc tcattaatac actctcccag ttctttttca ccgactccaa cggcgtgatt 960
atgagcttca gcaagtctct gctgcatgac ctcatgctgt acttttttga ccataagggc 1020
gaactgacca acaaggagct gagcagcttt ctcctcaagc atggactgga gctgaacagc 1080
aaagagaaat ccaataaaaa gtatagactg aattatatga aacaactcac agatagcacc 1140
atctttgaga ccgagatgat cgccagcttc agagaggaga tcgagacctc cagctatagg 1200
agcgtgaact ctctgagcaa taagatcggc aactgcatcg gccagttcat tacccctctc 1260
aagagaaagg aggagctgac caacatcctc atcgacacca attaccccaa ggagctggct 1320
tccaagctgg ccgacagcat taaggtgatc aaaagccaga gcgtcgccaa tatttccaac 1380
aaatatatgc tcgaagccat tcacgctttt gagtccggca agaagtacgg cgacttccaa 1440
gccgaattca acgagacaag agagctggag gaccaccatt ttatgaagaa taacaagctg 1500
atcgcctttc aagacagcga tctgatcaga aaccccgtgg tctatagaac cattaaccaa 1560
tctagaaaaa tcattaacgc cgccatcaat aaatataata ttgtcagaat caacatcgag 1620
gtcgccagcg atgtgaacaa gtccttcgaa caaagagata acgacaagaa gtaccagaac 1680
gacaactatg agaaaaatct gcagctggag tccgagctca ccgactatat caataaggaa 1740
aatctccacg tcaacgtgaa cagcaaaatg atggagagat acaaactgta cctcagccaa 1800
aataagcact gcatctatac aaacacccct ctgacaatga tggacgtgat ctatagcacc 1860
aacgtgcaag tggatcacat cattccccag tccaagattc tggatgacac cctcaacaat 1920
aaggtcctcg tgctcagaga cgccaactcc atcaaaaaca atagactccc tctggaggcc 1980
ttcgatgaga tgcagatcaa tgtggatacc aactacacaa aaaaggacta tctcaccgag 2040
tgcctccatc tgctgaagaa caaaaccaat cccatcagca aaaaaaagta tcagtacctc 2100
accctcaaga agctggacga tgagacaatt gagggcttta ttagcagaaa tattaacgac 2160
acaaggtaca ttaccagata cattgccaat tatctgaaga ccgctttcaa agagagcgac 2220
aagacaaaga atattgatgt cgtgacaatt aaaggcgccg tgacctctag atttagaaag 2280
aggtggctga ccacatatga cgagtatgga taccacccca ccatctattc tctggaggac 2340
aagggaagga acctctacta ctaccaccac gctatcgacg ccatcattct cgccaacatt 2400
gacaagaggt acattacact ggccaacgcc tacgatacca ttaggctgat taagatcgat 2460
agaaatctgt ccaaggagca gaagcaaagg gacatcgaca cagtcatcaa aaatacagtc 2520
aaaagcatga gcaagtacca tggcttctcc gaggactaca ttagaagcct catgagcaag 2580
aaccacattc ccgccatctg caagaacctc tccgatgaag tgcagatcag aatccccctc 2640
aagttcaaca ccgactatga caacctcggc tacagattca ccgatgacca gtatcactat 2700
aaaaagctgt acattgcctt caaggaggcc cagaatgccc tcaaagaaaa ggaaaccctc 2760
gaaaaggaac tgatcgagag gtttaataac gaggctcaaa tcctcaacgc taatatcatt 2820
ctgacctaca ccggcttcga gtccaacaat gaactgatcg acatcaagaa ggccaagaag 2880
gtgaccgaca cactgaagcc taatctcaag aattacatca aggccattga tatcctcacc 2940
caagaggaat ataccaagag atgtctggag tactataatg actccgagtt cgccacccag 3000
ctcaagattc cttacgtgaa tttcaaaatt aataaaaggt ttaggggcaa gatccaaggc 3060
agcgaaaacg ccgtctctct gagagaggtg ctgaagaaga caaagctgaa cagcttcgaa 3120
gagtttgagt cctatctcaa gagcgaagac ggcattaagt ccccctacta catcaaatat 3180
acaaagaaca cactgggcaa ggagtcctac accatctacg aggctaactc ctactactgt 3240
gccgagatct acaccgatag ccagaataag ccccagctga gaggaattag atacgtcgac 3300
gtgagaaaag aggacggcaa actcgtgctg ctgaagcccc tcccctccac atgcaagcat 3360
atcacctatc tcttccataa tgaatatatt gctatttata aagacagcaa ttataagagg 3420
ctcaagaata acggcttcgg agcttataga agcatcaaca acgtgaacgt gaataagatt 3480
attattagac tgtttgccaa tcagaacctc aacgacaacg acgtggtcat caccagcagc 3540
atcttcatca agaagtactc cctcgatgtc ttcggccaca tcaatggcga gatcaagtgc 3600
ggcgaccagt ctctgttcac cattaagaaa aga 3633
<210> 141
<211> 3633
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<223> 人源化的MAD2007 V2
<400> 141
atggagaact accgccagaa gcaccggttt gtgctggcca ccgatctggg catcggcagc 60
aatggctggg ccatcatcga cctggatgca cacagggtgg aggacctggg agtgcagatc 120
ttcgagtctg gcgaggaggg cgccaagaag gccagcgcca gggcctccca gcagcggaga 180
ctgaagagaa gcgcccacag gctgaacagg cgcaagaagc agagaaagga ggccctgatc 240
aagtttctgc aggagatcga gttccccgac ctggtggaga tcctgaactc ttttaagaag 300
cagaagaacc ctaatgatat cctgagcctg agggtgaagg gcctggacaa taagctgagc 360
cccctggagc tgttttccat cctgatctac atgtctaaca atcgcggcta taaggatttc 420
tacgacaacg atatcaatga caacaatacc gacaaggatg agaaggagat ggagaaggcc 480
aagtccacca tcgagaagct gtttgccagc aactcctaca ggacagtggg cgagatgatc 540
gccaccgacc caacattcat cgtggataag tctggcagca agaaagtgat caagtatcac 600
aacaagaagg gctatcagta cctgatcccc cgcaagctgc tggagaatga gatgtctctg 660
atcctgcaca agcaggagga gttttacgac tgcctgagca tcgataatat caccatcatc 720
ctggataaga tcttctttca gcggaatttc gaggacggcc ccggccctaa gaacaagaga 780
gacgattaca agaacaattc taagggcaac cagttttata ccggcttcaa tgagatgatc 840
ggcctgtgcc ctttctaccc aaacgagaag aagggcacaa agaatagcct gatctatgat 900
gagtactatc tgatcaatac cctgagccag ttctttttca cagactccaa cggcgtgatc 960
atgtcctttt ctaagagcct gctgcacgac ctgatgctgt actttttcga tcacaagggc 1020
gagctgacca acaaggagct gagctccttc ctgctgaagc acggcctgga gctgaattct 1080
aaggagaaga gcaacaagaa gtatcggctg aattacatga agcagctgac cgactccaca 1140
atctttgaga ccgagatgat cgcctctttc agggaggaga tcgagacatc tagctaccgc 1200
agcgtgaact ccctgtctaa taagatcggc aactgcatcg gccagtttat cacccctctg 1260
aagagaaagg aggagctgac caacatcctg atcgatacaa attatccaaa ggagctggcc 1320
agcaagctgg ccgacagcat caaagtgatc aagagccagt ccgtggccaa catcagcaat 1380
aagtacatgc tggaggccat ccacgccttc gagtccggca agaagtatgg cgattttcag 1440
gccgagttca acgagaccag ggagctggag gaccaccact ttatgaaaaa caataagctg 1500
atcgccttcc aggactccga tctgatcaga aatcctgtgg tgtacaggac aatcaaccag 1560
tctcgcaaga tcatcaatgc cgccatcaac aagtataata tcgtgcggat caacatcgag 1620
gtggcctctg acgtgaataa gagcttcgag cagagagaca acgataagaa gtaccagaac 1680
gataattatg agaagaacct gcagctggag agcgagctga ccgactacat caacaaggag 1740
aatctgcacg tgaacgtgaa tagcaagatg atggagcggt acaagctgta cctgtcccag 1800
aacaagcact gcatctacac caatacaccc ctgaccatga tggatgtgat ctattccaca 1860
aacgtgcagg tggaccacat catccctcag tctaagatcc tggacgatac actgaacaat 1920
aaggtgctgg tgctgcggga tgccaattcc atcaagaaca atagactgcc tctggaggcc 1980
tttgacgaga tgcagatcaa tgtggatacc aactacacaa agaaggacta tctgaccgag 2040
tgtctgcacc tgctgaagaa caagacaaat ccaatcagca agaagaagta tcagtacctg 2100
accctgaaga agctggacga tgagacaatc gagggcttca tctccagaaa catcaatgac 2160
acccgctata tcacacggta catcgccaac tatctgaaga ccgcctttaa ggagtccgac 2220
aagacaaaga atatcgatgt ggtgaccatc aagggcgccg tgacatctcg gttcagaaag 2280
aggtggctga ccacatacga tgagtatggc taccacccaa ccatctatag cctggaggac 2340
aagggcagga acctgtacta ttaccaccac gccatcgatg ccatcatcct ggccaatatc 2400
gacaagcggt acatcaccct ggccaacgcc tatgatacaa tccggctgat caagatcgac 2460
agaaacctgt ccaaggagca gaagcagcgg gacatcgata ccgtgatcaa gaatacagtg 2520
aagtctatga gcaagtacca cggcttttcc gaggattata tcagatccct gatgtctaag 2580
aatcacatcc cagccatctg caagaacctg tctgacgagg tgcagatcag gatccccctg 2640
aagtttaata ccgactacga taacctgggc tatcgcttca cagacgatca gtatcactac 2700
aagaagctgt acatcgcctt caaggaggcc cagaacgccc tgaaggagaa ggagaccctg 2760
gagaaggagc tgatcgagcg gtttaacaat gaggcccaga tcctgaacgc caatatcatc 2820
ctgacctata caggcttcga gagcaacaat gagctgatcg atatcaagaa ggccaagaag 2880
gtgaccgaca cactgaagcc aaacctgaag aattacatca aggccatcga tatcctgacc 2940
caggaggagt atacaaagcg gtgtctggag tattacaacg actccgagtt cgccacccag 3000
ctgaagatcc cctacgtgaa cttcaagatc aacaagcggt tccggggcaa gatccagggc 3060
tccgagaacg ccgtgtctct gagagaggtg ctgaagaaga caaagctgaa ttcttttgag 3120
gagttcgagt cctacctgaa gtctgaggat ggcatcaaga gcccctatta catcaagtat 3180
accaagaata cactgggcaa ggagtcttat accatctacg aggccaacag ctattactgc 3240
gccgagatct acacagactc ccagaataag cctcagctga gaggcatcag atatgtggac 3300
gtgagaaagg aggatggcaa gctggtgctg ctgaagccac tgccctctac ctgtaagcac 3360
atcacatacc tgtttcacaa tgagtacatc gccatctata aggacagcaa ctacaagagg 3420
ctgaagaaca atggctttgg cgcctatcgc tccatcaaca atgtgaacgt gaataagatc 3480
atcatcaggc tgttcgccaa ccagaatctg aacgacaatg acgtggtcat cacctcctct 3540
atctttatca agaagtacag cctggacgtg ttcggccaca tcaacggcga gatcaagtgt 3600
ggcgaccaga gcctgttcac catcaagaag cgg 3633
<210> 142
<211> 1211
<212> PRT
<213> 阿扎夏普氏菌(Sharpea azabuensis)
<400> 142
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Asp Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ala Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Val Glu Ile Leu Asn
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Ile Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Asp Asn Asn Thr Asp Asn Asp Glu Lys Glu Met Gln Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Ile Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Ile Thr Asn Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Lys Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Ser Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Ile Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Gly Thr Asn Val Gln Val
610 615 620
Asp His Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Ile Leu Glu Lys Glu Leu Thr Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Ile Asp Thr Leu Lys Pro Asp Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Glu Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Lys Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210
<210> 143
<211> 1211
<212> PRT
<213> 阿扎夏普氏菌(Sharpea azabuensis)
<400> 143
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Asp Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ser Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Asn Asn Ile Leu Asp
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Val Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Glu Asp Lys Lys Asp Ser Asp Glu Lys Glu Met Gln Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Ile Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Leu Thr Asn Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Arg Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Leu Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Phe Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Gly Thr Asn Val Gln Val
610 615 620
Asp His Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Ile Leu Glu Lys Glu Leu Thr Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Ile Asp Thr Leu Lys Pro Asp Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Glu Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Lys Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210
<210> 144
<211> 1211
<212> PRT
<213> 阿扎夏普氏菌(Sharpea azabuensis)
<400> 144
Met Glu Asn Tyr Arg Gln Lys His Arg Phe Val Leu Ala Thr Asp Leu
1 5 10 15
Gly Ile Gly Ser Asn Gly Trp Ala Ile Ile Asp Leu Asp Ala His Arg
20 25 30
Val Glu Asp Leu Gly Val Gln Ile Phe Glu Ser Gly Glu Glu Gly Ala
35 40 45
Lys Lys Ala Ser Ala Arg Ala Ser Gln Gln Arg Arg Leu Lys Arg Ser
50 55 60
Ala His Arg Leu Asn Arg Arg Lys Lys Gln Arg Lys Glu Ala Leu Ile
65 70 75 80
Lys Phe Leu Gln Glu Ile Glu Phe Pro Asp Leu Val Glu Ile Leu Asn
85 90 95
Ser Phe Lys Lys Gln Lys Asn Pro Asn Asp Ile Leu Ser Leu Arg Val
100 105 110
Lys Gly Leu Asp Asn Lys Leu Ser Pro Leu Glu Leu Phe Ser Val Leu
115 120 125
Ile Tyr Met Ser Asn Asn Arg Gly Tyr Lys Asp Phe Tyr Asp Asn Asp
130 135 140
Ile Asn Glu Asp Lys Lys Asp Ser Asp Glu Lys Glu Met Gln Lys Ala
145 150 155 160
Lys Ser Thr Ile Glu Lys Leu Phe Ala Ser Asn Ser Tyr Arg Thr Val
165 170 175
Gly Glu Met Ile Ala Thr Asp Pro Thr Phe Ile Val Asp Lys Ser Gly
180 185 190
Ser Lys Lys Val Ile Lys Tyr His Asn Lys Lys Gly Tyr Gln Tyr Leu
195 200 205
Ile Pro Arg Lys Leu Leu Glu Asn Glu Met Ser Leu Ile Leu His Lys
210 215 220
Gln Glu Glu Phe Tyr Asp Cys Leu Ser Ile Asp Asn Val Thr Ile Ile
225 230 235 240
Leu Asp Lys Ile Phe Phe Gln Arg Asn Phe Glu Asp Gly Pro Gly Pro
245 250 255
Lys Asn Lys Arg Asp Asp Tyr Lys Asn Asn Ser Lys Gly Asn Gln Phe
260 265 270
Tyr Thr Gly Phe Asn Glu Met Ile Gly Leu Cys Pro Phe Tyr Pro Asn
275 280 285
Glu Lys Lys Gly Thr Lys Asn Ser Leu Ile Tyr Asp Glu Tyr Tyr Leu
290 295 300
Ile Asn Thr Leu Ser Gln Phe Phe Phe Thr Asp Ser Asn Gly Val Ile
305 310 315 320
Met Ser Phe Ser Lys Ser Leu Leu His Asp Leu Met Leu Tyr Phe Phe
325 330 335
Asp His Lys Gly Glu Leu Thr Tyr Lys Glu Leu Ser Ser Phe Leu Leu
340 345 350
Lys His Gly Leu Glu Leu Asn Ser Lys Glu Lys Ser Asn Lys Lys Tyr
355 360 365
Arg Leu Asn Tyr Met Lys Gln Leu Thr Asp Ser Thr Ile Phe Glu Thr
370 375 380
Glu Met Ile Ala Ser Phe Arg Glu Glu Ile Glu Thr Ser Ser Tyr Arg
385 390 395 400
Ser Val Asn Ser Leu Ser Asn Lys Ile Gly Asn Cys Ile Gly Gln Phe
405 410 415
Ile Thr Pro Leu Lys Arg Lys Glu Glu Leu Thr Asn Ile Leu Ile Asp
420 425 430
Thr Asn Tyr Pro Lys Glu Leu Ala Ser Lys Leu Ala Asp Ser Ile Lys
435 440 445
Val Ile Lys Ser Gln Ser Val Ala Asn Ile Ser Asn Lys Tyr Met Leu
450 455 460
Glu Ala Ile His Ala Phe Glu Ser Gly Lys Lys Tyr Gly Asp Phe Gln
465 470 475 480
Ala Glu Phe Asn Glu Thr Arg Glu Leu Glu Asp His His Phe Met Lys
485 490 495
Asn Asn Lys Leu Ile Ala Phe Gln Asp Ser Asp Leu Ile Arg Asn Pro
500 505 510
Val Val Tyr Arg Thr Ile Asn Gln Ser Arg Lys Ile Ile Asn Ala Ala
515 520 525
Ile Asn Lys Tyr Asn Ile Val Arg Ile Asn Ile Glu Val Ala Ser Asp
530 535 540
Val Asn Lys Ser Phe Glu Gln Arg Asp Asn Asp Lys Lys Tyr Gln Asn
545 550 555 560
Asp Asn Tyr Glu Lys Asn Leu Gln Leu Glu Ser Glu Leu Thr Asp Tyr
565 570 575
Ile Asn Lys Glu Asn Leu His Val Asn Val Asn Ser Lys Met Met Glu
580 585 590
Arg Tyr Lys Leu Tyr Leu Ser Gln Asn Lys His Cys Ile Tyr Thr Asn
595 600 605
Thr Pro Leu Thr Met Met Asp Val Ile Tyr Ser Thr Asn Val Gln Val
610 615 620
Asp His Ile Ile Pro Gln Ser Lys Ile Leu Asp Asp Thr Leu Asn Asn
625 630 635 640
Lys Val Leu Val Leu Arg Asp Ala Asn Ser Ile Lys Asn Asn Arg Leu
645 650 655
Pro Leu Glu Ala Phe Asp Glu Met Gln Ile Asn Val Asp Thr Asn Tyr
660 665 670
Thr Lys Lys Asp Tyr Leu Thr Glu Cys Leu His Leu Leu Lys Asn Lys
675 680 685
Thr Asn Pro Ile Ser Lys Lys Lys Tyr Gln Tyr Leu Thr Leu Lys Lys
690 695 700
Leu Asp Asp Glu Thr Ile Glu Gly Phe Ile Ser Arg Asn Ile Asn Asp
705 710 715 720
Thr Arg Tyr Ile Thr Arg Tyr Ile Ala Asn Tyr Leu Lys Thr Ala Phe
725 730 735
Lys Glu Ser Asp Lys Thr Lys Asn Ile Asp Val Val Thr Ile Lys Gly
740 745 750
Ala Val Thr Ser Arg Phe Arg Lys Arg Trp Leu Thr Thr Tyr Asp Glu
755 760 765
Tyr Gly Tyr His Pro Thr Ile Tyr Ser Leu Glu Asp Lys Gly Arg Asn
770 775 780
Leu Tyr Tyr Tyr His His Ala Ile Asp Ala Ile Ile Leu Ala Asn Ile
785 790 795 800
Asp Lys Arg Tyr Ile Thr Leu Ala Asn Ala Tyr Asp Thr Ile Arg Leu
805 810 815
Ile Lys Ile Asp Arg Asn Leu Ser Lys Glu Gln Lys Gln Arg Asp Ile
820 825 830
Asp Thr Val Ile Lys Asn Thr Val Lys Ser Met Ser Lys Tyr His Gly
835 840 845
Phe Ser Glu Asp Tyr Ile Arg Ser Leu Met Ser Lys Asn His Ile Pro
850 855 860
Ala Ile Cys Lys Asn Leu Ser Asp Glu Val Gln Ile Arg Ile Pro Leu
865 870 875 880
Lys Phe Asn Thr Asp Tyr Asp Asn Leu Gly Tyr Arg Phe Thr Asp Asp
885 890 895
Gln Tyr His Tyr Lys Lys Leu Tyr Ile Ala Phe Lys Glu Ala Gln Asn
900 905 910
Ala Leu Lys Glu Lys Glu Thr Leu Glu Lys Glu Leu Ile Glu Arg Phe
915 920 925
Asn Asn Glu Ala Gln Ile Leu Asn Ala Asn Ile Ile Leu Thr Tyr Thr
930 935 940
Gly Phe Glu Ser Asn Asn Glu Leu Ile Asp Ile Lys Lys Ala Lys Lys
945 950 955 960
Val Thr Asp Thr Leu Lys Pro Asn Leu Lys Asn Tyr Ile Lys Ala Ile
965 970 975
Asp Ile Leu Thr Gln Glu Glu Tyr Thr Lys Arg Cys Leu Glu Tyr Tyr
980 985 990
Asn Asp Ser Glu Phe Ala Thr Gln Leu Lys Ile Pro Tyr Val Asn Phe
995 1000 1005
Lys Ile Asn Lys Arg Phe Arg Gly Lys Ile Gln Gly Ser Glu Asn
1010 1015 1020
Ala Val Ser Leu Arg Glu Val Leu Lys Lys Thr Lys Leu Asn Ser
1025 1030 1035
Phe Glu Glu Phe Glu Ser Tyr Leu Lys Ser Glu Asp Gly Ile Lys
1040 1045 1050
Ser Pro Tyr Tyr Ile Lys Tyr Thr Lys Asn Thr Leu Gly Lys Glu
1055 1060 1065
Ser Tyr Thr Ile Tyr Glu Ala Asn Ser Tyr Tyr Cys Ala Glu Ile
1070 1075 1080
Tyr Thr Asp Ser Gln Asn Lys Pro Gln Leu Arg Gly Ile Arg Tyr
1085 1090 1095
Val Asp Val Arg Lys Glu Asp Gly Lys Leu Val Leu Leu Lys Pro
1100 1105 1110
Leu Pro Ser Thr Cys Lys His Ile Thr Tyr Leu Phe His Asn Glu
1115 1120 1125
Tyr Ile Ala Ile Tyr Lys Asp Ser Asn Tyr Lys Arg Leu Lys Asn
1130 1135 1140
Asn Gly Phe Gly Ala Tyr Arg Ser Ile Asn Asn Val Asn Val Asn
1145 1150 1155
Lys Ile Ile Ile Arg Leu Phe Ala Asn Gln Asn Leu Asn Asp Asn
1160 1165 1170
Asp Val Val Ile Thr Ser Ser Ile Phe Ile Lys Lys Tyr Ser Leu
1175 1180 1185
Asp Val Phe Gly His Ile Asn Gly Glu Ile Lys Cys Gly Asp Gln
1190 1195 1200
Ser Leu Phe Thr Ile Lys Lys Arg
1205 1210

Claims (6)

1.一种核酸酶系统，所述核酸酶系统被配置为执行核酸引导的核酸酶编辑，其中所述核酸酶系统选自包括SEQ ID No.7(MAD2007核酸酶)、SEQ ID No.26(gRNA重复序列)和SEQID No.27(gRNA tracr序列)的核酸酶系统；包括SEQ ID No.7(MAD2007核酸酶)、SEQ IDNo.28(gRNA重复序列)和SEQ ID No.29(gRNA tracr序列)的核酸酶系统；以及包括SEQ IDNo.7(MAD2007核酸酶)、SEQ ID No.30(gRNA重复序列)和SEQ ID No.31(gRNA tracr序列)的核酸酶系统。
2.根据权利要求1所述的核酸酶系统，所述核酸酶系统包括SEQ ID No.7(MAD2007核酸酶)、SEQ ID No.26(gRNA重复序列)和SEQ ID No.27(gRNA tracr序列)。
3.根据权利要求1所述的核酸酶系统，所述核酸酶系统包括SEQ ID No.7(MAD2007核酸酶)、SEQ ID No.28(gRNA重复序列)和SEQ ID No.29(gRNA tracr序列)。
4.根据权利要求1所述的核酸酶系统，所述核酸酶系统包括SEQ ID No.7(MAD2007核酸酶)、SEQ ID No.30(gRNA重复序列)和SEQ ID No.31(gRNA tracr序列)。
5.一种切口酶，所述切口酶具有选自SEQ ID No.17和SEQ ID No.18的氨基酸序列。
6.一种失活核酸酶，所述失活核酸酶具有选自SEQ ID No.19的氨基酸序列。
CN202080086100.4A 2019-12-10 2020-04-01 新颖的mad核酸酶 Active CN114787347B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202410829829.6A CN118638870A (zh) 2019-12-10 2020-04-01 新颖的mad核酸酶

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201962946282P 2019-12-10 2019-12-10
US62/946,282 2019-12-10
PCT/US2020/026095 WO2021118626A1 (en) 2019-12-10 2020-04-01 Novel mad nucleases

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CN202410829829.6A Division CN118638870A (zh) 2019-12-10 2020-04-01 新颖的mad核酸酶

Publications (2)

Publication Number Publication Date
CN114787347A true CN114787347A (zh) 2022-07-22
CN114787347B CN114787347B (zh) 2024-07-12

Family

ID=74045155

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202410829829.6A Pending CN118638870A (zh) 2019-12-10 2020-04-01 新颖的mad核酸酶
CN202080086100.4A Active CN114787347B (zh) 2019-12-10 2020-04-01 新颖的mad核酸酶

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN202410829829.6A Pending CN118638870A (zh) 2019-12-10 2020-04-01 新颖的mad核酸酶

Country Status (7)

Country Link
US (6) US10883095B1 (zh)
EP (1) EP4069837A4 (zh)
KR (1) KR20220110778A (zh)
CN (2) CN118638870A (zh)
AU (1) AU2020402526A1 (zh)
CA (1) CA3157131A1 (zh)
WO (1) WO2021118626A1 (zh)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11208649B2 (en) 2015-12-07 2021-12-28 Zymergen Inc. HTP genomic engineering platform
US9988624B2 (en) 2015-12-07 2018-06-05 Zymergen Inc. Microbial strain improvement by a HTP genomic engineering platform
WO2022125329A1 (en) * 2020-12-07 2022-06-16 Inscripta, Inc. gRNA STABILIZATION IN NUCLEIC ACID-GUIDED NICKASE EDITING
AU2021415461A1 (en) * 2021-01-04 2023-08-17 Inscripta, Inc. Mad nucleases
US11332742B1 (en) * 2021-01-07 2022-05-17 Inscripta, Inc. Mad nucleases
US11884924B2 (en) * 2021-02-16 2024-01-30 Inscripta, Inc. Dual strand nucleic acid-guided nickase editing
IL308806A (en) 2021-06-01 2024-01-01 Arbor Biotechnologies Inc Gene editing systems including nuclease crisper and their uses
WO2023028521A1 (en) 2021-08-24 2023-03-02 Inscripta, Inc. Genome-wide rationally-designed mutations leading to enhanced cellobiohydrolase i production in s. cerevisiae
AU2023216314A1 (en) 2022-02-02 2024-08-22 Inscripta, Inc. Nucleic acid-guided nickase fusion proteins
US20240052370A1 (en) 2022-07-27 2024-02-15 Inscripta, Inc. Modulating cellular repair mechanisms for genomic editing
WO2024062138A1 (en) 2022-09-23 2024-03-28 Mnemo Therapeutics Immune cells comprising a modified suv39h1 gene
US20240301447A1 (en) 2023-02-15 2024-09-12 Arbor Biotechnologies, Inc. Gene editing method for inhibiting aberrant splicing in stathmin 2 (stmn2) transcript

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170240922A1 (en) * 2014-02-11 2017-08-24 The Regents of University of Colorado, a body Corporate Crispr enabled multiplexed genome engineering
CN107532161A (zh) * 2015-03-03 2018-01-02 通用医疗公司 具有改变的PAM特异性的工程化CRISPR‑Cas9核酸酶
CN107849546A (zh) * 2015-05-15 2018-03-27 先锋国际良种公司 对cas内切核酸酶系统、pam序列和指导rna元件的快速表征
CN107922931A (zh) * 2015-06-12 2018-04-17 普拉克生物化学公司 热稳定的Cas9核酸酶
CN108350449A (zh) * 2015-08-28 2018-07-31 通用医疗公司 工程化的CRISPR-Cas9核酸酶
US20190367949A1 (en) * 2018-06-05 2019-12-05 Lifeedit, Inc. Rna-guided nucleases and active fragments and variants thereof and methods of use

Family Cites Families (77)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6391582B2 (en) 1998-08-14 2002-05-21 Rigel Pharmaceuticlas, Inc. Shuttle vectors
SE9900530D0 (sv) 1999-02-15 1999-02-15 Vincenzo Vassarotti A device for concentrating and/or purifying macromolecules in a solution and a method for manufacturing such a device
US7087415B2 (en) 2000-07-31 2006-08-08 Athena Biotechnologies, Inc. Methods and compositions for directed gene assembly
US20020139741A1 (en) 2001-03-27 2002-10-03 Henry Kopf Integral gasketed filtration cassette article and method of making the same
US7166443B2 (en) 2001-10-11 2007-01-23 Aviva Biosciences Corporation Methods, compositions, and automated systems for separating rare cells from fluid samples
AU2003251286B2 (en) 2002-01-23 2007-08-16 The University Of Utah Research Foundation Targeted chromosomal mutagenesis using zinc finger nucleases
AU2003268089A1 (en) 2002-08-13 2004-02-25 National Jewish Medical And Research Center Method for identifying mhc-presented peptide epitopes for t cells
DK2325332T3 (da) 2005-08-26 2013-01-28 Dupont Nutrition Biosci Aps Anvendelse af CRISPR-associerede gener (CAS)
WO2009025690A2 (en) 2007-05-23 2009-02-26 Nature Technology Corporation Improved e. coli plasmid dna production
GB0724860D0 (en) 2007-12-20 2008-01-30 Heptares Therapeutics Ltd Screening
US8450112B2 (en) 2008-04-09 2013-05-28 Maxcyte, Inc. Engineering and delivery of therapeutic compositions of freshly isolated cells
US20100076057A1 (en) 2008-09-23 2010-03-25 Northwestern University TARGET DNA INTERFERENCE WITH crRNA
EP2206723A1 (en) 2009-01-12 2010-07-14 Bonas, Ulla Modular DNA-binding domains
GB0922434D0 (en) 2009-12-22 2010-02-03 Ucb Pharma Sa antibodies and fragments thereof
WO2011072246A2 (en) 2009-12-10 2011-06-16 Regents Of The University Of Minnesota Tal effector-mediated dna modification
EA024121B9 (ru) 2010-05-10 2017-01-30 Дзе Реджентс Ов Дзе Юниверсити Ов Калифорния Композиции эндорибонуклеаз и способы их использования
EP2395087A1 (en) 2010-06-11 2011-12-14 Icon Genetics GmbH System and method of modular cloning
US9361427B2 (en) 2011-02-01 2016-06-07 The Regents Of The University Of California Scar-less multi-part DNA assembly design automation
US8332160B1 (en) 2011-11-17 2012-12-11 Amyris Biotechnologies, Inc. Systems and methods for engineering nucleic acid constructs using scoring techniques
US9637739B2 (en) 2012-03-20 2017-05-02 Vilnius University RNA-directed DNA cleavage by the Cas9-crRNA complex
PT2800811T (pt) 2012-05-25 2017-08-17 Univ California Métodos e composições para modificação de adn alvo dirigida por arn e para modulação dirigida por arn de transcrição
CN113512577A (zh) 2012-06-25 2021-10-19 Gen9股份有限公司 用于核酸组装和高通量测序的方法
EP3494997B1 (en) 2012-07-25 2019-09-18 The Broad Institute, Inc. Inducible dna binding proteins and genome perturbation tools and applications thereof
ES2757325T3 (es) 2012-12-06 2020-04-28 Sigma Aldrich Co Llc Modificación y regulación del genoma en base a CRISPR
US20140310830A1 (en) * 2012-12-12 2014-10-16 Feng Zhang CRISPR-Cas Nickase Systems, Methods And Compositions For Sequence Manipulation in Eukaryotes
US8697359B1 (en) 2012-12-12 2014-04-15 The Broad Institute, Inc. CRISPR-Cas systems and methods for altering expression of gene products
WO2014143381A1 (en) 2013-03-09 2014-09-18 Agilent Technologies, Inc. Methods of in vivo engineering of large sequences using multiple crispr/cas selections of recombineering events
US9499855B2 (en) 2013-03-14 2016-11-22 Elwha Llc Compositions, methods, and computer systems related to making and administering modified T cells
WO2014144495A1 (en) 2013-03-15 2014-09-18 Abvitro, Inc. Single cell bar-coding for antibody discovery
US9234213B2 (en) 2013-03-15 2016-01-12 System Biosciences, Llc Compositions and methods directed to CRISPR/Cas genomic engineering systems
EP4286517A3 (en) 2013-04-04 2024-03-13 President and Fellows of Harvard College Therapeutic uses of genome editing with crispr/cas systems
US9776138B2 (en) 2013-06-25 2017-10-03 Tetra Laval Holdings & Finance S.A. Membrane filtration device having a hygienic suspension arrangement
KR20220025922A (ko) 2013-07-09 2022-03-03 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 멀티플렉스 rna-가이드된 게놈 조작
US10822606B2 (en) 2013-09-27 2020-11-03 The Regents Of The University Of California Optimized small guide RNAs and methods of use
US20150098954A1 (en) 2013-10-08 2015-04-09 Elwha Llc Compositions and Methods Related to CRISPR Targeting
US10767156B2 (en) 2013-10-24 2020-09-08 Yeda Research And Development Co., Ltd. Polynucleotides encoding BREX system polypeptides and methods of using same
BR112016013400B1 (pt) 2013-12-11 2023-02-14 Regeneron Pharmaceuticals, Inc. Método in vitro para modificar um genoma em um lócus genômico de interesse em uma célula pluripotente
CN106103705A (zh) 2013-12-12 2016-11-09 布罗德研究所有限公司 核苷酸重复障碍中crispr‑cas系统的组合物和使用方法
US10787654B2 (en) 2014-01-24 2020-09-29 North Carolina State University Methods and compositions for sequence guiding Cas9 targeting
WO2015143558A1 (en) 2014-03-27 2015-10-01 British Columbia Cancer Agency Branch T-cell epitope identification
WO2015153940A1 (en) 2014-04-03 2015-10-08 Massachusetts Institute Of Technology Methods and compositions for the production of guide rna
KR101815695B1 (ko) 2014-05-28 2018-01-08 기초과학연구원 표적 특이적 뉴클레아제를 이용한 표적 dna의 민감한 검출 방법
US20160053272A1 (en) 2014-07-18 2016-02-25 Whitehead Institute For Biomedical Research Methods Of Modifying A Sequence Using CRISPR
US20160053304A1 (en) 2014-07-18 2016-02-25 Whitehead Institute For Biomedical Research Methods Of Depleting Target Sequences Using CRISPR
WO2016014900A2 (en) 2014-07-25 2016-01-28 Novogy, Inc. Promoters derived from yarrowia lipolytica and arxula adeninivorans, and methods of use thereof
US20160076093A1 (en) 2014-08-04 2016-03-17 University Of Washington Multiplex homology-directed repair
CN107002078A (zh) 2014-10-09 2017-08-01 生命技术公司 Crispr寡核苷酸和基因剪辑
WO2016110453A1 (en) 2015-01-06 2016-07-14 Dsm Ip Assets B.V. A crispr-cas system for a filamentous fungal host cell
EP3271382B1 (en) 2015-03-16 2020-02-12 Max-Delbrück-Centrum für Molekulare Medizin in der Helmholtz-Gemeinschaft Method of detecting new immunogenic t cell epitopes and isolating new antigen-specific t cell receptors by means of an mhc cell library
US11608511B2 (en) 2015-04-13 2023-03-21 Maxcyte, Inc. Methods for modifying genomic DNA
US9790490B2 (en) 2015-06-18 2017-10-17 The Broad Institute Inc. CRISPR enzymes and systems
WO2017009399A1 (en) 2015-07-13 2017-01-19 Institut Pasteur Improving sequence-specific antimicrobials by blocking dna repair
CN108369230B (zh) 2015-09-25 2021-09-17 阿布维特罗有限责任公司 用于对天然配对t细胞受体序列进行t细胞受体靶向鉴别的高通量方法
WO2017078631A1 (en) 2015-11-05 2017-05-11 Agency For Science, Technology And Research Chemical-inducible genome engineering technology
WO2017083722A1 (en) 2015-11-11 2017-05-18 Greenberg Kenneth P Crispr compositions and methods of using the same for gene therapy
US9988624B2 (en) 2015-12-07 2018-06-05 Zymergen Inc. Microbial strain improvement by a HTP genomic engineering platform
US20200263165A1 (en) 2015-12-18 2020-08-20 Danisco Us Inc. Methods and compositions for polymerase ii (pol-ii) based guide rna expression
EP3402890A1 (en) 2016-01-12 2018-11-21 SQZ Biotechnologies Company Intracellular delivery of complexes
EP3199632A1 (en) 2016-01-26 2017-08-02 ACIB GmbH Temperature-inducible crispr/cas system
US9896696B2 (en) 2016-02-15 2018-02-20 Benson Hill Biosystems, Inc. Compositions and methods for modifying genomes
EP3430407B1 (en) 2016-03-18 2021-10-06 QT Holdings Corp Kit, composition, device and method for cell separation
CA3017678A1 (en) 2016-04-04 2017-10-12 Eth Zurich Mammalian cell line for protein production and library generation
CN109715803B (zh) 2016-04-25 2023-07-07 巴塞尔大学 等位基因编辑及其应用
PT3272867T (pt) 2016-06-02 2019-12-04 Sigma Aldrich Co Llc Utilização de proteínas de ligação ao dna programáveis para intensificar a modificação de genoma direcionada
US10988779B2 (en) 2016-06-22 2021-04-27 Icahn School Of Medicine At Mount Sinai Viral delivery of RNA utilizing self-cleaving ribozymes and CRISPR-based applications thereof
CA3029254A1 (en) 2016-06-24 2017-12-28 The Regents Of The University Of Colorado, A Body Corporate Methods for generating barcoded combinatorial libraries
US20190264193A1 (en) 2016-08-12 2019-08-29 Caribou Biosciences, Inc. Protein engineering methods
CN109715804A (zh) 2016-09-23 2019-05-03 帝斯曼知识产权资产管理有限公司 用于宿主细胞的指导rna表达系统
EP3526326A4 (en) * 2016-10-12 2020-07-29 The Regents of The University of Colorado, A Body Corporate NEW MODIFIED AND CHEMERICAL NUCLEASES
DK3535290T5 (da) 2016-11-07 2024-09-02 Genovie Ab Manipuleret todelt celleindretning til opdagelse og karakterisering af t-celle-receptorinteraktion med beslægtet antigen
KR20190133699A (ko) * 2017-03-24 2019-12-03 큐어백 아게 Crispr-연관 단백질을 암호화하는 핵산 및 이의 용도
US20180362590A1 (en) 2017-04-14 2018-12-20 Synthetic Genomics, Inc. Polypeptides with type v crispr activity and uses thereof
US10011849B1 (en) 2017-06-23 2018-07-03 Inscripta, Inc. Nucleic acid-guided nucleases
NZ760730A (en) * 2017-06-23 2023-04-28 Inscripta Inc Nucleic acid-guided nucleases
US9982279B1 (en) 2017-06-23 2018-05-29 Inscripta, Inc. Nucleic acid-guided nucleases
EP3848459A1 (en) 2017-06-30 2021-07-14 Inscripta, Inc. Automated cell processing methods, modules, instruments and systems
KR20200124702A (ko) * 2018-02-23 2020-11-03 파이어니어 하이 부렛드 인터내쇼날 인코포레이팃드 신규한 cas9 오르소로그

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170240922A1 (en) * 2014-02-11 2017-08-24 The Regents of University of Colorado, a body Corporate Crispr enabled multiplexed genome engineering
CN107532161A (zh) * 2015-03-03 2018-01-02 通用医疗公司 具有改变的PAM特异性的工程化CRISPR‑Cas9核酸酶
CN107849546A (zh) * 2015-05-15 2018-03-27 先锋国际良种公司 对cas内切核酸酶系统、pam序列和指导rna元件的快速表征
CN107922931A (zh) * 2015-06-12 2018-04-17 普拉克生物化学公司 热稳定的Cas9核酸酶
CN108350449A (zh) * 2015-08-28 2018-07-31 通用医疗公司 工程化的CRISPR-Cas9核酸酶
US20190367949A1 (en) * 2018-06-05 2019-12-05 Lifeedit, Inc. Rna-guided nucleases and active fragments and variants thereof and methods of use

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
HAFT, D.H.等: "type II CRISPR RNA-guided endonuclease Cas9 [Enterococcus faecalis]", 《NCBI：WP_033789179.1》, pages 1 *

Also Published As

Publication number Publication date
US20220049233A1 (en) 2022-02-17
EP4069837A1 (en) 2022-10-12
AU2020402526A1 (en) 2022-06-09
US10883095B1 (en) 2021-01-05
CN118638870A (zh) 2024-09-13
EP4069837A4 (en) 2024-03-13
US20210324355A1 (en) 2021-10-21
CA3157131A1 (en) 2021-06-17
US11193115B2 (en) 2021-12-07
US11085030B2 (en) 2021-08-10
US11053485B2 (en) 2021-07-06
US20210171923A1 (en) 2021-06-10
US20210301270A1 (en) 2021-09-30
WO2021118626A1 (en) 2021-06-17
KR20220110778A (ko) 2022-08-09
US11174471B2 (en) 2021-11-16
CN114787347B (zh) 2024-07-12
US20210198641A1 (en) 2021-07-01

Similar Documents

Publication Publication Date Title
CN114787347B (zh) 新颖的mad核酸酶
US11345903B2 (en) Engineered enzymes
EP4069281A1 (en) Novel enzymes
US20220213458A1 (en) Mad nucleases
US11332742B1 (en) Mad nucleases
US20240376451A1 (en) Mad nucleases
US11214781B2 (en) Engineered enzyme
US12146170B2 (en) Engineered enzyme
US20240052370A1 (en) Modulating cellular repair mechanisms for genomic editing

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