CN110382690A - 新型逆转录酶及其用途 - Google Patents

新型逆转录酶及其用途 Download PDF

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CN110382690A
CN110382690A CN201880012198.1A CN201880012198A CN110382690A CN 110382690 A CN110382690 A CN 110382690A CN 201880012198 A CN201880012198 A CN 201880012198A CN 110382690 A CN110382690 A CN 110382690A
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龚小松
王岩
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Abstract

提供了从FLVRT和MLVRT的部分形成的杂合逆转录酶。

Description

新型逆转录酶及其用途
相关专利申请的交叉引用
本专利申请要求2017年2月16日提交的美国临时专利申请号62/459,974的优先权，其通过引用纳入本文用于所有目的。
关于通过EFS-WEB以ASCII文本文件形式提交的“序列表”
根据37C.F.R.§§1.821-1.825，在机型IBM-PC，MS-Windows操作系统上，创建于2018年2月2日的184,742字节的文件094260-1076461-112710PC_SequenceListing.txt的序列表通过引用全文纳入本文用于所有目的。
背景技术
核酸的检测，分析，转录和扩增是现代分子生物学中经常使用的程序。DNA聚合酶可用于检测和扩增DNA或RNA。这种RNA分析程序的应用可能涉及基因表达的研究，传染原或遗传疾病的诊断以及cDNA的产生，仅举几例应用。RNA的逆转录因此具有许多用途。在一些情况下，逆转录之后是聚合酶链式反应扩增，其可以用于RNA的快速检测和定量。
发明概述
在一些实施方式中，提供了杂合逆转录酶，其包含指(finger)结构域，掌(palm)结构域，拇指(thumb)结构域，连接结构域和RNA酶H结构域，其中至少一个所述结构域是小鼠白血病病毒逆转录酶(MLVRT)而其他所述结构域来自猫白血病病毒逆转录酶(FLVRT)。
在一些实施方式中，杂合逆转录酶包含：小鼠白血病病毒逆转录酶(MLVRT)的部分，其包含与包含拇指，连接和RNA酶H结构域的猫白血病病毒逆转录酶(FLVRT)的部分连接的指和掌结构域。在一些实施方式中，MLVRT的部分具有羧基末端，并且羧基末端直接连接至FLVRT的部分的氨基末端。在一些实施方式中，MLVRT的部分具有羧基末端，并且羧基末端通过1-100个氨基酸的连接氨基酸序列连接至FLVRT的部分的氨基末端。在一些实施方式中，MLVRT的部分与SEQ ID NO：1至少95％相同，并且FLVRT的部分与SEQ ID NO：5至少95％相同。
在一些实施方式中，MLVRT的部分包含SEQ ID NO：1，并且FLVRT的部分包含SEQ IDNO：5。在一些实施方式中，FLVRT的部分包含SEQ ID NO：6或SEQ ID NO：10。在一些实施方式中，FLVRT的部分包含SEQ ID NO：7，SEQ ID NO：8，SEQ ID NO：9或SEQ ID NO：11。
在一些实施方式中，MLVRT的部分包含SEQ ID NO：2。在一些实施方式中，MLVRT的部分包含SEQ ID NO：3或SEQ ID NO：4。
在一些实施方式中，MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ IDNO：7；或
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：8；或
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：9；或
MLVRT的部分包含SEQ ID NO：3，并且FLVRT的部分包含SEQ ID NO：11；或
MLVRT的部分包含SEQ ID NO：3，并且FLVRT的部分包含SEQ ID NO：10；或
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：13。
在一些实施方式中，杂合逆转录酶包含与SEQ ID NO：14，15，16，17，18，19，34或35基本上(例如，至少70％，80％，85％，90％或95％)相同的序列。
在一些实施方案中，杂合逆转录酶包含：猫白血病病毒逆转录酶(FLVRT)的部分，其包含与包含RNA酶H结构域的小鼠白血病病毒逆转录酶(MLVRT)的部分连接的指和掌结构域。在一些实施方式中，MLVRT的部分包括拇指，连接和RNA酶H结构域。在一些实施方式中，FLVRT的部分包含与SEQ ID NO：26至少95％相同的序列；并且MLVRT的部分包含与SEQ IDNO：28至少95％相同的序列。在一些实施方式中，杂合逆转录酶包含与SEQ ID NO：30基本上(例如，至少70％，80％，85％，90％或95％)相同的序列。
在一些实施方式中，FLVRT的部分包括指，掌，拇指和连接结构域。在一些实施方式中，FLVRT的部分包含与SEQ ID NO：27基本上(例如，至少70％，80％，85％，90％或95％)相同的序列；并且MLVRT的部分包含与SEQ ID NO：29基本上(例如，至少70％，80％，85％，90％或95％)相同的序列。在一些实施方式中，杂合逆转录酶包含与SEQ ID NO：31基本上(例如，至少70％，80％，85％，90％或95％)相同的序列。
在一些实施方式中，如上文或本文其他地方所述的杂合逆转录酶具有改善热稳定性的至少一个突变。在一些实施方式中，所述至少一个突变位于对应于SEQ ID NO：34中的L139，D200，N479，D522，F526，H592，L601，E605和H632的位置处。
还提供了核酸，其包含编码如上文或本文其他地方所述的杂合逆转录酶的多核苷酸。在一些实施方式中，所述核酸还包含与多核苷酸操作性地连接的(任选地异源的)启动子。
还提供表达载体，其含有如上文或本文他处所述的核酸。还提供一种细胞，其包含所述表达载体。在一些实施方式中，细胞是细菌细胞。
还提供了一种反应混合物，其包含：RNA或DNA模板；和如上文或本文其他地方所述的杂合逆转录酶。在一些实施方式中，反应混合物还包含至少一种寡核苷酸引物和/或脱氧核苷酸。
还提供了进行逆转录的方法。在一些实施方式中，该方法包括将如上文或本文其他地方所述的杂合逆转录酶在一定条件下与模板RNA和引物接触，所述引物与模板RNA杂交，所述条件使杂合逆转录酶以模板RNA依赖性方式延伸引物，以形成cDNA。在一些实施方式中，条件包括42-60℃之间的延伸步骤。
还提供了包含如上文或本文其他地方所述的杂合逆转录酶的试剂盒。在一些实施方式中，试剂盒还包括DNA聚合酶。
定义
除非另有说明，本文所用的所有科技术语具有本发明所属领域普通技术人员通常所理解的含义。通常，本文所用的命名和下述细胞培养、分子遗传学、有机化学和核酸化学以及杂交中的实验室步骤均为本领域熟知和常用的。使用标准技术进行核酸和肽合成。按照本领域和各种通用参考文献所述的常规方法进行这些技术和步骤(通常参见，Sambrook等，《分子克隆：实验室手册》(MOLECULAR CLONING：A LABORATORY MANUAL)，第2版(1989)冷泉港实验室出版社(Cold Spring Harbor Laboratory Press)，纽约冷泉港(Cold SpringHarbor，N.Y.)，其通过引用纳入本文)，将它们引入本文全文中。本文所用的命名以及下述分析化学和有机合成中的实验室步骤均为本领域熟知且常用。也可使用标准技术或其变型进行化学合成或化学分析。
用于蛋白质部分时，“异源”指该蛋白质包含在天然情况下彼此不存在于同一关系中的两个或更多结构域。这样的蛋白质，例如融合蛋白，例如本文所述的杂合RT，含有通过肽键或肽接头序列共价连接的两个或更多个序列，以排列成新的功能性蛋白质。
“引物”指与靶核酸上的序列杂交并且用作核酸合成的起始点的多核苷酸序列。引物可以是各种长度的并且通常长度小于50个核苷酸，例如长度为12-30个核苷酸。可基于本领域技术人员已知的原理设计用于PCR的引物的长度和序列，参见例如Innis等(同上)。
“聚合酶”是指进行模板引导的多核苷酸合成的酶。该术语同时包括全长多肽和具有聚合酶活性的结构域。
“模板”指多核苷酸序列，其包含任选侧接一个或两个引物杂交位点的待扩增的多核苷酸。
本文所用的“核酸”表示DNA、RNA、单链、双链、或更高度聚集的杂交基序及其任意化学修饰。修饰包括但不限于，提供引入其它电荷、极化性、氢键、静电相互作用、与核酸配体碱基或核酸配体整体的连接点和作用点的化学基团的那些修饰。这类修饰包括但不限于，肽核酸(PNA)、磷酸二酯基团修饰(例如，硫代磷酸酯、甲基膦酸酯)、2′-位糖修饰、5-位嘧啶修饰、8-位嘌呤修饰、环外胺处的修饰、4-硫尿核苷的取代、5-溴或5-碘-尿嘧啶的取代、骨架修饰、甲基化、不常见的碱基配对组合如异碱基(isobase)、异胞苷和异胍(isoguanidine)等。核酸也可包含非天然碱基，如硝基吲哚。修饰还可包括3′和5′修饰，例如用荧光团(例如，量子点)或其它部分加帽。
术语“多肽”、“肽”和“蛋白质”在本文中互换使用，指氨基酸残基的聚合物。该术语适用于其中一个或多个氨基酸残基是相应天然产生氨基酸的人造化学模拟物的氨基酸聚合物，以及天然产生的氨基酸聚合物和非天然产生的氨基酸聚合物。
术语“氨基酸”指天然产生的和合成的氨基酸，以及作用方式类似于天然产生氨基酸的氨基酸类似物和氨基酸模拟物。天然产生的氨基酸是由遗传密码编码的那些氨基酸，以及随后经修饰的那些氨基酸，例如，羟基脯氨酸、γ-羧基谷氨酸和O-磷酸丝氨酸。氨基酸类似物指与天然产生的氨基酸具有相同基本化学结构的化合物，即碳原子与氢原子、羧基、氨基和R基结合，如高丝氨酸、正亮氨酸、甲硫氨酸亚砜、甲硫氨酸甲基锍。这种类似物具有修饰的R基(如正亮氨酸)或修饰的肽主链，但保留了与天然产生的氨基酸基本相同的化学结构。氨基酸模拟物指结构不同于氨基酸的普通化学结构，但作用方式类似于天然产生氨基酸的化合物。
在本文中，氨基酸可用IUPAC-IUB生物化学命名委员会推荐的通用三字母符号或单字母符号表示。同样，核苷酸可由其公认的单字母代码表示。
术语“启动子”指定位于转录起始上游和/或下游且涉及识别和结合RNA聚合酶和其它蛋白质以起始转录的区域或序列。
“载体”指多核苷酸，其独立于宿主染色体时能够在宿主生物体中复制。优选的载体包括质粒且通常具有复制起始点。载体可包括，例如，转录和翻译终止子、转录和翻译起始序列和用于调控特定核酸表达的启动子。
当如下述比对最大对应性时，如果在两条序列中核苷酸或氨基酸残基的序列分别是相同的，两条核酸序列或多肽被称为“相同的”。在两条或更多条核酸或多肽序列内容中的术语“相同的”或“相同性”百分比指就比较窗的最大对应性进行比较和比对时，相同或有特定百分数的相同氨基酸残基或核苷酸的两条或更多条序列或子序列，如使用下列序列比较算法之一或通过手工比对和目测测量。序列相同性百分比涉及蛋白和肽使用时，认为不同的残基位置通常由于保守氨基酸取代而不同，其中氨基酸残基被具有相似化学性质(如电荷或疏水性)的其他氨基酸残基取代，因此不改变分子的功能性质。当序列由于保守取代而不同时，序列相同性百分比可上调以根据该取代的保守性质校正。进行该调整的方法为本领域技术人员公知。通常其包括将保守取代作为部分评分而不是完全错配，从而增加序列相同性百分比。因此例如，相同氨基酸得分为1，非保守取代得分为0，保守取代得分为0-1。保守取代的评分按照，例如Meyers和Miller的算法，Computer Applic.Biol.Sci.4：11-17(1988)，例如，如在程序PC/GENE(美国加利福尼亚州芒廷维尤的智慧遗传公司(Intelligenetics，Mountain View，California，USA))中执行的进行计算。
如果采用以下序列比较算法之一或通过手工比对和目测检查测量时，当就比较窗口或指定区域上的最大对应性而言，这些序列有特定百分数的核苷酸或氨基酸残基相同(例如在指定区域上或未指定整个序列(SEQ ID NO)时至少60％、70％、75％、80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％、99％相同)，则序列彼此间“基本相同”。
为了序列比较，一般将一个序列用作与测试序列比较的参比序列。当使用序列比较算法时，将测试和参比序列输入计算机，如果需要，指定子序列坐标，并指定序列算法程序参数。可使用默认的程序参数，或者可指定另外的参数。然后，序列比较算法基于程序参数计算测试序列相对于参比序列的序列相同性百分比。
本文所用的“比较窗口”包括参考选自20-600，通常约50-200，更常见约100-150的任一数量的毗连位置的区段，其中对两个序列进行最优比对后，可将序列与具有相同数量毗连位置的参比序列作比较。比对序列的比较方法是本领域熟知的。可进行最优序列比对以作比较，例如，通过Smith和Waterman的局部同源性算法，Adv.Appl.Math.2：482(1981)；通过Needleman和Wunsch的同源性比对算法，J.Mol.Biol.48：443(1970)；通过Pearson和Lipman的相似性搜索法，Proc.Nat’l.Acad.Sci.USA85：2444(1988)；通过计算机执行这些算法(阿克赛勒里公司(Accelrys)的威斯康星遗传学软件包(Wisconsin GeneticsSoftware Package)中的GAP、BESTFIT、FASTA和TFASTA)，或通过手工比对和目测。
使用BLAST算法确定百分比序列相同性和序列相似性，其在Altschul等，J.Mol.Biol.215：403-410(1990)中描述。进行BLAST分析的软件可从国家生物技术信息中心(National Center for Biotechnology Information)公开获得(http：//ncbi.nlm.nih.gov/)。此算法包括：首先通过鉴定查询序列中长度为W的短字来鉴定高评分序列对(HSP)，与数据库序列中长度相同的字比对时它们能匹配或满足一些正值的阈值评分T。T称为相邻字评分阈值(Altschul等，同上)。这些初始相邻字命中(word hit)用作启动搜索的种子，以便找到含有它们的较长HSP。只要可提高累积比对评分，该字命中在两个方向上沿各序列延伸。出现以下情况时中止字命中在各个方向上的延伸：累积比对评分比其最大获得值降低X；由于一个或多个负评分残基比对的累积，累积评分变为零或零以下；或者达到任一序列的末端。BLAST算法参数W、T和X确定该比对的灵敏度和速度。BLAST程序使用的默认值为：字长(W)11，BLOSUM62评分矩阵(参见Henikoff和Henikoff，Proc.Natl.Acad.Sci.USA89：10915(1989))比对(B)50，期望值(E)10，M＝5，N＝-4，以及比较两条链。
附图说明
图1描绘了RT突变体表达后的示例性可溶性级分SDS-PAGE凝胶。
发明详述
发明人已经发现，与猫白血病病毒逆转录酶(FLVRT)酶相比，由小鼠白血病病毒逆转录酶(MLVRT)和FLVRT形成的逆转录酶(RT)杂合体具有改善的溶解性。与非杂合MLVRT或FLVRT酶中的至少一种相比，预期本文所述的杂合体也具有改善的稳定性，表达或其组合。例如，本发明人已经产生了杂合逆转录酶，其包含指结构域，掌结构域，拇指结构域，连接结构域和RNA酶H结构域，其中至少一个所述结构域是小鼠白血病病毒逆转录酶(MLVRT)而其他所述结构域来自猫白血病病毒逆转录酶(FLVRT)。如更详细讨论的，本发明人已经产生了杂合体，其中指和掌结构域是FLVRT或MLVRT序列，其中至少一些剩余部分来自替代的酶。
多肽
本文提供了具有五个RT结构域(从氨基至羧基：指，掌，拇指，连接和RNA酶H结构域)的杂合逆转录酶(RT)，其中这些结构域中的至少一个(例如，1、2、3或4个)来自MLVRT而其余结构域来自FLVRT。MLVRT和指，掌，拇指，连接和RNA酶H结构域的结构描述于例如Das和Georgiadis，Structure12：819-829(2004)中。与其中所有结构域均来自FLVRT(即野生型FLVRT)的RT相比，所得杂合RT具有改善的表达(例如，在大肠杆菌中)，而在一些实施方式中，与MLVRT相比具有改善的准确性。
MLV-FLV杂合体
在一些实施方式中，杂合RT包含与FLVRT的拇指，连接和RNA酶H结构域连接的MLVRT的指和掌结构域。包含指和掌结构域的MLVRT的示例性部分包括，例如SEQ ID NO：1或其基本相同的序列。在一些实施方式中，包含指和掌结构域的MLVRT部分包含SEQ ID NO：2或与其基本相同的序列。上述MLVRT部分可以连接到包含拇指，连接和RNA酶H结构域的FLVRT的部分。包含拇指，连接和RNA酶H结构域的FLVRT的示例性部分是SEQ ID NO：5或其基本相同的序列。在一些实施方式中，包含拇指，连接和RNA酶H结构域的FLVRT的部分是SEQID NO：6或其基本相同的序列或SEQ ID NO：10或其基本相同的序列。示例性杂合RT可包含，例如SEQ ID NO：1或SEQ ID NO：2，SEQ ID NO：5，SEQ ID NO：6或SEQ ID NO：10中任一。在一些实施方式中，杂合RT包含SEQ ID NO：14，15，16，17，18或19之一或其基本相同的序列。
FLV-MLV杂合体
在一些实施方式中，杂合RT至少包含FLVRT的指和掌结构域，其与FLVRT或MLVRT的拇指和连接结构域连接，而其转而与MLVRT的RNA酶H结构域连接。在一些实施方式中，杂合RT包含FLVRT的指和掌结构域以及MLVRT的拇指，连接和RNA酶H结构域。包含指和掌结构域的FLVRT的示例性部分包括，例如SEQ ID NO：26或其基本相同的序列。包含拇指，连接和RNA酶H结构域的MLVRT的示例性部分包括，例如SEQ ID NO：28或其基本相同的序列。
在一些实施方式中，杂合RT包含FLVRT的指，掌，拇指和连接结构域以及MLVRT的RNA酶H结构域。包含指，掌，拇指和连接结构域的FLVRT的示例性部分包括，例如SEQ ID NO：27或其基本相同的序列。包含RNA酶H结构域的MLVRT的示例性部分包括，例如SEQ ID NO：29或其基本相同的序列。在一些实施方式中，杂合RT包含SEQ ID NO：30或31之一或其基本相同的序列。
本文描述的任何杂合RT可以在氨基或羧基末端包含其他氨基酸。示例性的其他氨基酸序列可以包括，例如，允许纯化蛋白质的表位标签或其他标签或其潜在密码子允许克隆位点的表位标签或其他标签。这类标签可以在杂交RT的任一端融合，因为最方便纯化。此类标签的示例包括但不限于多组氨酸序列或FLAG标签。还可以包括各种接头序列以将这类标签或其他序列与杂合RT序列连接。接头可以包括例如，甘氨酸，丝氨酸或其他不显著干扰蛋白质折叠的氨基酸，从而不损害杂合RT的活性。接头序列还可以包括蛋白酶切割序列，使得任选地一旦纯化了杂合RT(例如，使用标签)，可以通过蛋白酶或其他切割机制除去标签。在一些实施方式中，杂合RT在羧基末端包含一个或多个(例如，2-20，2-5，例如3个)丙氨酸。
热稳定突变
如本文所述，本文所述的任何杂合RT可包括一种或多种突变，其改善酶的热稳定性(即，在暴露于超过37℃的温度，例如42-60℃期间或之后保留活性的能力)。示例性突变包括在对应于SEQ ID NO：34中的以下位置处的一个或多个(例如，2，3，4，5，6或更多个)突变：L139(包括但不限于L139P)，D200(包括但不限于D200N)，N479(包括但不限于N479D)，D522(包括但不限于D522G，D522N或D522A)，F526(包括但不限于F526I)，H592(包括但不限于H592K)，L601(包括但不限于L601W)，E605(包括但不限于E605K)和H632(包括但不限于H632Y)。应理解，这类位置名称不表示权利要求所要保护的分子本身中氨基酸的编号，而是表示权利要求所要保护的分子序列与SEQ ID NO：34最大程度对齐时权利要求所要保护的分子中的残基位置。
部分的连接
如本文所述的不同RT的两个部分可以通过本领域技术人员熟知的方法通过接头接合。这些方法可包括重组和化学方法。
连接不同RT的部分也可包括在通过标准肽合成化学或重组方法分开合成的基团之间形成肽键。或者，在一些实施方式中，杂合RT中各部分的编码序列直接接合并表达为融合蛋白。或者，氨基酸接头序列也可编码在多肽编码序列中并用于将第一和第二RT部分分开足够的距离以确保每个多肽折叠成其二级和三级结构。使用本领域熟知的重组技术将这类氨基酸接头序列整合至融合蛋白中。合适的肽接头序列可基于以下因素选择：(1)其采取挠性延伸构象的能力；(2)其采取可与第一和第二多肽上功能性表位相互作用的二级结构的能力；以及(3)缺少可与多肽功能性表位反应的疏水性或带电残基。典型的肽接头序列含有Gly、Ser、Val和Thr残基。其他的近中性氨基酸(如Ala)也可用于接头序列。可用作接头的氨基酸序列包括下述中公开的那些：Maratea等，(1985)Gene40：39-46；Murphy等，(1986)Proc.Natl.Acad.Sci.USA83：8258-8262；美国专利号4,935,233和4,751,180。该接头序列的长度通常可以是1至约50个氨基酸，例如长度为3、4、6或10个氨基酸，但长度也可以是100或200个氨基酸。不一定需要接头序列。
化学连接可以例如如《生物偶联技术》(Bioconjugate Techniques)，Hermanson编，学术出版社(Academic Press)(1996)中所述进行。接合可包括，例如，出于将2种蛋白质互相连接的目的的衍生化，所述衍生化通过蛋白质化学领域所熟知的方法以直接或通过连接化合物的方式进行。例如，在一个化学偶联实施方式中，连接催化结构域和核酸结合结构域的手段包括杂双官能-偶联剂，其最终导致在2个基团之间形成分子间二硫键。可用于本发明的这种能力的其他类型的偶联剂描述于，例如，美国专利号4,545,985。或者，分子间二硫键可在各基团中的半胱氨酸之间方便地形成，其是天然产生的或者通过遗传工程改造插入的。连接基团的手段也可使用双官能交联剂或特定低pH可切割交联剂之间的硫醚连接或特定蛋白酶可切割接头或其他可切割或不可切割化学连接。其他化学接头包括碳水化合物接头、脂质接头、脂肪酸接头、聚醚接头，如PEG，等。例如，聚(乙二醇)接头可获自阿拉巴马州汉茨维尔的谢氏聚合物有限公司(Shearwater Polymers，Inc.)。这些接头任选具有酰胺键、巯基键或杂双官能键。连接基团可以是化学交联剂，包括，例如琥珀酰亚胺基-(N-马来酰亚胺甲基)-环己烷-1-羧酸酯(SMCC)。连接基团也可以是其他氨基酸序列，包括但不限于，例如，聚丙氨酸、聚甘氨酸或类似的连接基团。
在一些实施方式中，可以取代或添加方式向序列中导入非经典氨基酸或化学氨基酸类似物。非经典氨基酸一般包括但不限于：普通氨基酸的D-异构体、α-氨基异丁酸、4-氨基丁酸、Abu、2-氨基丁酸、γ-Abu、e-Ahx、6-氨基己酸、Aib、2-氨基异丁酸、3-氨基丙酸、鸟氨酸、正亮氨酸、正缬氨酸、羟基脯氨酸、肌氨酸、瓜氨酸、磺基丙氨酸、叔丁基甘氨酸、叔丁基丙氨酸、苯基甘氨酸、环己基丙氨酸、β-丙氨酸、氟代-氨基酸、设计氨基酸如β-甲基氨基酸、Cα-甲基氨基酸、Nα-甲基氨基酸和氨基酸类似物。此外，氨基酸可以是D(右旋)或L(左旋)。
表达和纯化
编码杂合RT的核酸可以使用重组遗传学领域的常规技术获得。公开本发明所用一般方法的基础文本包括Sambrook和Russell，Molecular Cloning，A Laboratory Manual(《分子克隆，实验室手册》)(第3版，2001)；Kriegler，Gene Transfer and Expression：ALaboratory Manual(《基因转移和表达：实验室手册》)(1990)；以及Current Protocols inMolecular Biology(《新编分子生物学实验指南》)(Ausubel等编，1994-1999))。这类核酸还可通过体外扩增方法获得，例如描述于以下文献中的那些：Berger，Sambrook，和Ausubel，以及Mullis等(1987)美国专利号4,683,202；PCR ProtocolsAGuide to Methodsand Applications(《PCR实验方法：方法和应用指南》)(Innis等编)学术出版社公司，加利福尼亚州圣地亚哥(1990)(Innis)；Arnheim和Levinson(1990年10月1日)C&EN36-47；TheJournal Of NIH Research(1991)3：81-94；Kwoh等(1989)Proc.Natl.Acad.Sci.USA86：1173；Guatelli等(1990)Proc.Natl.Acad.Sci.USA87，1874；Lomell等(1989)J.Clin.Chem.，35：1826；Landegren等(1988)Science 241：1077-1080；Van Brunt(1990)Biotechnology 8：291-294；Wu和Wallace(1989)Gene4：560；以及Barringer等(1990)Gene89：117，其各自通过引用全文纳入本文用于所有目的且尤其是用于所有涉及扩增方法的教导。
技术人员将认识到可以对杂合RT进行另外的修饰而不降低其生物活性。可进行一些修饰以促进克隆、表达或将结构域整合至融合蛋白中。这类修饰为本领域技术人员所熟知并且包括，例如，在编码结合结构域的多核苷酸的一端添加密码子(例如在氨基端添加甲硫氨酸)以提供起始位点，或将额外的氨基酸(如聚His)置于一端以建立方便地设置的限制性酶切位点或终止密码子或纯化序列。
本发明所述的杂合RT多肽可在多种宿主细胞中表达，包括大肠杆菌、其他细菌宿主、酵母、丝状真菌和多种高级真核细胞如COS、CHO和HeLa细胞系和骨髓瘤细胞系。在微生物中进行基因表达的技术描述于例如Smith，Gene Expression in RecombinantMicroorganisms(重组微生物中的基因表达)(Bioprocess Technology(《生物加工技术》)，卷22)，Marcel Dekker，1994。可用于表达的细菌的示例包括但不限于，埃希菌属，肠杆菌，固氮菌属，欧文氏菌属，杆菌属，假单胞菌属，克雷伯氏菌属，变形杆菌属，沙门氏菌属，沙雷氏菌属，志贺氏菌属，根瘤菌属，透明颤菌属和副球菌属。可用作表达宿主的丝状真菌包括例如下列属：曲霉属，木霉属，链孢霉属，青霉属，头孢霉属，绵霉属，柄孢壳菌属，毛霉属，旋孢腔菌属和梨孢属。参见，例如，美国专利号5,679,543以及Stahl和Tudzynski编，《丝状真菌分子生物学》(Molecular Biology in Filamentous Fungi)，约翰韦利父子公司(JohnWiley&Sons)，1992。在酵母中合成异源蛋白质是众所周知的并描述于文献中。Sherman，F等，《酵母遗传方法》(Methods in Yeast Genetics)，冷泉港实验室(1982)是一项熟知的工作，描述了可用于在酵母中产生酶的各种方法。
存在许多本领域普通技术人员已知的生成多肽的表达系统。(参见例如，GeneExpression Systems(《基因表达系统》)Fernandex和Hoeffler编，学术出版社，1999；Sambrook和Russell，同上；以及Ausubel等，同上)。通常，编码多肽的多核苷酸位于在所需宿主细胞中具有功能的启动子的控制下。许多不同的启动子是可用的且是本领域技术人员已知的，且可用于本发明的表达载体中，这取决于具体应用。通常，选择的启动子依赖于其中启动子将具有活性的细胞。还可任选地包括其他表达控制序列，如核糖体结合位点、转录终止位点等。包含一种或多种这些控制序列的构建体称作“表达盒”。因此，编码接合的多肽的核酸经整合以在所需宿主细胞中高水平表达。
通常通过克隆在特定细胞中表达的基因来将获得适用于特定宿主细胞的表达控制序列。通常使用的原核控制序列(其在本发明中定义为包含用于转录起始的启动子(任选具有操纵子)以及核糖体结合位点序列)包括这类通常使用的启动子作为β-内酰胺酶(青霉素酶)和乳糖(1ac)启动子系统(Change等，Nature(1977)198：1056)、色氨酸(trp)启动子系统(Goeddel等，Nucleic Acids Res.(1980)8：4057)、tac启动子(DeBoer等，Proc.Natl.Acad.Sci.U.S.A.(1983)80：21-25)；以及λ衍生的PL启动子和N基因核糖体结合位点(Shimatake等，Nature(1981)292：128)。具体的启动子系统并不重要，可使用在原核细胞中具有功能并提供所需活性水平的任何可用的启动子。标准的细菌表达载体包括质粒，如基于pBR322的质粒(如pBLUESCRIPTTM)、pSKF、pET23D、λ噬菌体衍生的载体和融合表达系统(如GST和LacZ)。还可向重组蛋白添加表位标签以提供方便的分离方法，例如c-myc、HA-标签、6-His标签、麦芽糖结合蛋白、VSV-G标签、抗DYKDDDDK(SEQ ID NO：42)标签或任何这类标签，许多标签是本领域技术人员熟知的。
本文所述的多肽可以在细胞内表达，或可以从细胞分泌。细胞内表达常常导致高产量。如果需要，可通过进行重折叠程序来增加可溶性活性融合多肽的量(参见例如Sambrook等，同上；Marston等，Bio/Technology(1984)2：800；Schoner等，Bio/Technology(1985)3：151)。多肽可在多种宿主细胞中表达，包括大肠杆菌、其他细菌宿主、酵母、和多种高级真核细胞如COS、CHO和HeLa细胞系和骨髓瘤细胞系。宿主细胞可以是哺乳动物细胞，昆虫细胞或微生物，例如酵母细胞，细菌细胞或真菌细胞。
一旦表达，可按照本领域的标准程序纯化多肽，包括硫酸铵沉淀，亲和柱，柱层析，凝胶电泳等(一般参见R.Scopes，“蛋白质纯化”(Protein Purification)，Springer-Verlag，N.Y.(1982)，Deutscher，《酶学方法》(Methods in Enzymology)卷182：《蛋白质纯化指南》(Guide to Protein Purification)，学术出版公司，纽约(1990))。优选至少约90-95％均一性的基本纯化的组合物，并且最优选98-99％或更高的均一性。一旦根据需要部分纯化或均一化，然后可以使用多肽(例如，作为用于抗体生产的免疫原)。
为了便于多肽的纯化，编码多肽的核酸还可以包括可用亲和结合试剂针对的表位或“标签”的编码序列。合适的表位的示例包括myc和V-5报道基因；可用于重组生产具有这些表位的融合多肽的表达载体是可商购的(例如，英杰(Invitrogen)(加利福尼亚州卡尔斯巴德)载体pcDNA3.1/Myc-His和pcDNA3.1/V5-His适于在哺乳动物细胞中表达)。适用于将标签附着到本发明的融合蛋白上的其他表达载体和相应的检测系统是本领域技术人员已知的，并且一些是可商购的(例如，“FLAG”(柯达公司(Kodak)，纽约州罗切斯特(RochesterN.Y.))。合适的标签的另一个示例是能够结合金属螯合亲和配体的聚组氨酸序列。通常使用六个相邻的组氨酸，尽管可以使用多于或少于六个。可用作聚组氨酸标签的结合部分的合适金属螯合亲和配体包括次氮基三乙酸(NTA)(Hochuli，E.(1990)“用金属螯合吸附剂纯化重组蛋白”，在《基因工程改造：原理和方法》(Gene Engineering：Principlesand Methods)中，J.K.Setlow编，普莱南出版社(Plenum Press)，纽约；可从凯捷(Qiagen(加利福尼亚州圣克拉丽塔(Santa Clarita，Calif.))商购获得))。
在生物表达或纯化后，杂合RT多肽可具有与组成多肽的天然构象基本上不同的构象。在这种情况下，可能需要或希望变性并还原多肽，然后使多肽重新折叠成优选的构象。减少和变性蛋白质和诱导重折叠的方法是本领域技术人员熟知的(参见Debinski等，(1993)J.Biol.Chem.268：14065-14070；Kreitman和Pastan(1993)Bioconjug.Chem.4：581-585；和Buchner等，(1992)Anal.Biochem.205：263-270)。例如，Debinski等描述了胍-DTE中包涵体蛋白的变性和还原。然后将蛋白质在含有氧化型谷胱甘肽和L-精氨酸的氧化还原缓冲液中重折叠。
V.使用方法
逆转录(RT)是将RNA复制成DNA的扩增方法。RT反应可以如本文所述用反应混合物进行。例如，本发明提供了在允许逆转录和产生第一链和任选的第二链cDNA的条件下使用如本文所述的杂合RT逆转录一种或多种RNA(包括例如，细胞中的所有RNA，例如制备cDNA文库)。RT反应可以用随机引物，寡聚dT或RNA特异性引物引发。RT反应的组分和条件通常是已知的。
如果需要，反应可以进一步包括RT-PCR。用于进行PCR测定的标准技术是本领域已知的(《PCR技术：DNA扩增的原理和应用》(Principles and Applications for DNAAmplification)(Erlich编，1989)；《PCR方案：方法和应用指南》(PCR Protocols：A Guideto Methods and Applications)(Innis，Gelfland，Sninsky，和White编，1990)；Mattila等，Nucleic Acids Res.19：4967(1991)；Eckert和Kunkel，《PCR方法和应用》(PCR Methodsand Applications)1：17(1991)；Wallace等，连接酶链反应(Ligase Chain Reaction)，在《DNA损伤和突变检测技术》(Technologies for Detection of DNA Damage andMutations)中，第307-322页(Pfiefer编，1996))。RT和PCR反应通常用于相同的测定中，并被称为RT-PCR。RT-PCR将RNA逆转录成DNA与随后在单个反应中进行DNA扩增反应结合在一起。最佳的逆转录，杂交和扩增条件将根据所使用的引物和靶标的序列组成和长度，以及实施者选择的实验方法而变化。可以使用各种指南来选择合适的引物序列和杂交条件(参见例如Sambrook等，《分子克隆：实验室手册》(Molecular Cloning：A LaboratoryManual)(第2版)(1989)；Ausubel，F.M.等编，(1999)《分子生物学简明方案》(Short Protocols inMolecular Biology)，第4版，约翰威利父子有限公司(John Wiley&Sons))；Ausubel，F.M.等编，(1999-2010)《新编分子生物学方案》(Current Protocols in Molecular Biology)，约翰威利父子有限公司)。
在一些实施方式中，本文所述的杂合RT在比通常使用的温度更高的温度下用于逆转录酶反应。因此，在实施方式中，本文所述的一些杂合RT可以在37℃或42℃或高于42℃的温度，例如42-60℃，43-55℃，45-56℃，45-65℃等温度下使用。较高温度的RT反应在模板RNA在部分或完全抑制逆转录的正常RT温度(例如37℃或42℃)下形成二级结构的情况下特别有用。
VI.反应混合物
提供了包含本文所述杂合RT多肽的反应混合物。反应混合物可包含，例如，靶核酸，例如，其中发生逆转录的RNA靶标。反应混合物可以包含合适的核苷酸(例如，脱氧核苷酸(dNTP)或双脱氧核苷酸)，并且在一些实施方式中，包含至少一种缓冲剂。示例性的缓冲剂可包括例如但不限于，Tris，HEPES，ACES，PIPES，MOPSO，BES，MOPS，TES，TAPSO，POPSO，BICINE，TAPS或AMPSO。反应混合物可以任选地包含一种或多种用作模板依赖性核酸延伸的引物的寡核苷酸，一种或多种用作探针的寡核苷酸(例如，与标记物例如猝灭剂，荧光染料等连接)，和/或双链DNA结合染料(例如SYBRGREEN)。在一些实施方式中，反应混合物将进一步包含单独的DNA依赖性DNA聚合酶。在一些实施方式中，反应混合物将进一步包含镁(Mg++)。
VII.试剂盒
在一个方面，提供了用于进行逆转录(和任选的环状扩增，例如PCR)反应的试剂盒。在一些实施方式中，此类试剂盒包括如本文所述的杂合RT，和任选的dNTP，以及至少一种缓冲剂。这类试剂盒也可包含稳定剂和其他添加剂以提高扩增反应的效率。此类试剂盒还可包括一种或多种引物(例如聚-T，随机六聚体或特异性引物)以及使用试剂盒组分进行逆转录反应的说明书。在一些实施方式中，试剂盒将进一步包含单独的DNA依赖性DNA聚合酶。
实施例
为了增加重组FLV RT在大肠杆菌细胞中的溶解度，用来自MLV RT的RT多肽序列的部分和来自FLV RT的RT序列的部分构建杂合RT。制备的杂合RT构建体在大肠杆菌细胞中表现出改善的溶解性，如图1所示。
杂合RT FM1(1/3FLV-2/3MLV RT)包括来自氨基酸1-279的FLV RT的N-末端序列，其包括指和掌结构域，以及来自氨基酸280-671的MLV RT的C-末端序列，其包括拇指，连接和RNA酶H结构域。
杂合RT FM2(2/3FLV-1/3MLV RT)包括来自氨基酸1-497的FLV RT的N-末端序列，其包括指，掌，拇指和连接结构域，以及来自氨基酸498-671的MLV RT的C-末端序列，其包括RNA酶H结构域。
杂合RT MF包括来自氨基酸1-277的MLV RT的N-末端序列，其包括指和掌结构域，以及来自氨基酸276-667的FLV RT的C-末端序列，其包括拇指，连接和RNA酶H结构域。
杂合RT MF(P)包括来自氨基酸1-221的MLV RT的N-末端序列，其包括指和掌结构域，以及来自氨基酸221-667的FLV RT的C-末端序列，其包括拇指，连接和RNA酶H结构域。
将点突变引入MF和MF(P)杂合RT中以改善酶性能。
表达重组RT构建体的方法：
用含有100∶1比例的表达质粒的BL21细胞过夜培养物接种新鲜LB肉汤。培养物在25℃下生长约6小时或直至OD 600nm＝0.6-0.8。将IPTG加至0.1mM，并在16℃下O/N生长16小时。通过离心后收集沉淀收获细胞。将细胞重悬于200ml裂解缓冲液中并通过超声处理裂解。离心细胞碎片并收集200μl上清液。将5μl样品和5μl上样缓冲液在PCR条中合并，并在95℃下加热5分钟。将6ul样品上样到SDS-PAGE凝胶上进行分析。示例性结果示于图1。
测试了杂合蛋白和突变蛋白的许多特征，这些特征在下表中部分总结(空白表明未测试活性)：
以下提供了示例性突变和杂合序列的列表：
1.NF5PNAIY(SEQ ID NO：20)
基于FP(M)-TCH(F)，其具有来自MLV RT的指和掌结构域和来自FLV RT的拇指，连接和RNA酶H结构域。突变以下位置处的氨基酸：L139P(160)，D200N(221)，D522A(543)，F526I(547)，H632Y(653)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
2.MF4GC(SEQ ID NO：21)
基于FP(M)-TCH(F)，其具有来自MLV RT的指和掌结构域和来自FLV RT的拇指，连接和RNA酶H结构域，和延伸的C-末端序列。突变以下位置处的氨基酸：L139P(160)，D200N(221)，D522G(543)，L601W(622)，E605K(626)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
3.MF4G(SEQ ID NO：22)
基于FP(M)-TCH(F)，其具有来自MLV RT的指和掌结构域和来自FLV RT的拇指，连接和RNA酶H结构域，没有延伸的C-末端序列。突变以下位置处的氨基酸：L139P(160)，D200N(221)，D522G(543)，L601W(622)，E605K(626)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
4.MF(P)4G(SEQ ID NO：23)
基于FP(M)PstI-TCH(F)，与MF4G相比，其具有来自MLV RT的较短的指和掌结构域序列。来自FLV RT的拇指，连接和RNA酶H结构域没有延伸的C-末端序列。突变以下位置处的氨基酸：L139P(160)，D200N(221)，D522G(543)，L601W(622)，E605K(626)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
5.MF(P)4NDKW(SEQ ID NO：24)
基于FP(M)PstI-TCH(F)，与MF4G相比，其具有来自MLV RT的较短的指和掌结构域序列。来自FLV RT的拇指，连接和RNA酶H结构域没有延伸的C-末端序列。突变以下位置处的氨基酸：D200N(221)，N479D(500)，H592K(613)，L601W(622)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
6.MF(P)4NGWN(SEQ ID NO：25)
基于FP(M)PstI-TCH(F)，与MF4G相比，其具有来自MLV RT的较短的指和掌结构域序列。来自FLV RT的拇指，连接和RNA酶H结构域没有延伸的C-末端序列。突变以下位置处的氨基酸：D200N(221)，D522G(543)，L601W(622)，D651N(672)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
7.NF5PNAIYC(SEQ ID NO：39)
基于FP(M)-TCH(F)，其具有来自MLV RT的指和掌结构域和来自FLV RT的拇指，连接和RNA酶H结构域，和延伸的C-末端序列。突变以下位置处的氨基酸：L139P(160)，D200N(221)，D522A(543)，F526I(547)，H632Y(653)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
8.MF4(P)GC(SEQ ID NO：40)
基于FP(M)PstI-TCH(F)，与MF4G相比，其具有来自MLV RT的较短的指和掌结构域序列。来自FLV RT的拇指，连接和RNA酶H结构域，具有延伸的C-末端序列。突变以下位置处的氨基酸：L139P(160)，D200N(221)，D522G(543)，L601W(622)，E605K(626)。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
9.FM1(1/3FLV-2/3MLV RT)(SEQ ID NO：30)
具有来自FLV RT的N末端序列的1/3(指和掌结构域)和来自MLV RT的C末端序列的2/3剩余部分(拇指，连接，RNA酶H结构域)的杂合RT。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
10.FM2(2/3FLV-1/3MLV RT)(SEQ ID NO：31)
具有来自FLV RT的N末端序列的2/3(指，掌，拇指，连接结构域)和来自MLV RT的C末端序列的1/3的剩余部分(仅RNA酶H结构域)的杂合RT。斜体表示来自MLV的序列；粗体文本表示来自FLV的序列。
11.野生型MLV RT序列(SEQ ID NO：32)
12.野生型FLV RT序列(SEQ ID NO：33)
序列概述：
SEQ ID NO：1：通用短MLV
SEQ ID NO：2：通用较长MLV
SEQ ID NO：3：特定短MLV
SEQ ID NO：4：特定较长MLV
SEQ ID NO：5：通用较短w/o c-末端延伸FLV
SEQ ID NO：6：有较长c-末端延伸FLV：通用
SEQ ID NO：7：较短w/o c-末端延伸FLV：522，526，632处的特定突变
SEQ ID NO：8：有较长c-末端延伸FLV：522，601，605处的特定突变
SEQ ID NO：9：较短w/o c-末端延伸FLV：522，601，605处的特定突变
SEQ ID NO：10：长N-末端w/o延伸FLV：通用
SEQ ID NO：11：长N-末端w/o c-末端延伸FLV：522，601，605处的特定突变
SEQ ID NO：12：长N-末端w/o c-末端延伸FLV：479，592，601处的特定突变
SEQ ID NO：13：长N-末端w/o c-末端延伸FLV：522，601，651处的特定突变
SEQ ID NO：14：NF5PNAIY没有前导或末端序列
SEQ ID NO：15：MF4GC没有前导或末端序列
SEQ ID NO：16：MF4G没有前导或末端序列
SEQ ID NO：17：MF4(P)G没有前导或末端序列
SEQ ID NO：18：MF(P)4NDKW没有前导或末端序列
SEQ ID NO：19：MF(P)4NGWN没有前导或末端序列
SEQ ID NO：20：NF5PNAIY有前导和末端序列
SEQ ID NO：21：MF4GC有前导和末端序列
SEQ ID NO：22：MF4G有前导和末端序列
SEQ ID NO：23：MF(P)4G有前导和末端序列
SEQ ID NO：24：MF(P)4NDKW有前导和末端序列
SEQ ID NO：25：MF(P)4NGWN有前导和末端序列
SEQ ID NO：26：1/3FLVN-末端
SEQ ID NO：27 2/3FLVN-末端
SEQ ID NO：28 2/3MLV C-末端
SEQ ID NO：29 1/3MLV C-末端
SEQ ID NO：30 FM1(1/3 FLV-2/3 MLV RT)
SEQ ID NO：31 FM2(2/3 FLV-1/3 MLV RT)
SEQ ID NO：32野生型MLV RT序列
SEQ ID NO：33野生型FLV RT序列
SEQ ID NO：34：NF5PNAIYC没有前导或末端序列
SEQ ID NO：35：MF4(P)GC没有前导或末端序列
SEQ ID NO：36：NF5PNAIYC MLVRT部分
SEQ ID NO：37：NF5PNAIYC FLVRT部分
SEQ ID NO：38：MF4(P)GC MLVRT部分
SEQ ID NO：39：MF4(P)GC FLVRT部分
SEQ ID NO：40：NF5PNAIYC有前导和末端序列
SEQ ID NO：41：MF4(P)GC有前导和末端序列
本文所述的实施例和实施方式仅用于说明目的，本领域技术人员应了解据此作出的各种修饰或改变，且它们包括在本申请的主旨和权益以及所附权利要求书的范围内。本文引用的所有发表物、专利和专利申请通过引用全文纳入本文以用于所有目的。
序列表
SEQ ID NO：1：通用短MLV：
SEQ ID NO：2：通用较长MLV：
SEQ ID NO：3：特定短MLV：
SEQ ID NO：4：特定较长MLV：
SEQ ID NO：S：通用较短w/o c-末端延伸FLV：
SEQ ID NO：6：有较长c-末端延伸FLV：通用
SEQ ID NO：7：较短w/o c-末端延伸FLV：特定522，526，632
SEQ ID NO：8：有较长c-末端延伸FLV：522，601，605
SEQ ID NO：9：较短w/o c-末端延伸FLV：特定522，601，605
SEQ ID NO：10：长N-末端w/o c-末端延伸FLV：通用
SEQ ID NO：11：长N-末端w/o c-末端延伸FLV：特定522，601，605
SEQ ID NO：12：长N-末端w/o c-末端延伸FLVo特定479，592，601
SEQ ID NO：13：长N-末端w/o c-末端延伸FLV：特定522，601，651
SEQ ID NO：14：NF5PNAIY没有前导或末端序列：
SEQ ID NO：15：MF4GC没有前导或末端序列：
SEQ ID NO：16：MF4G没有前导或末端序列：
SEQ ID NO：17：MF4(P)G没有前导或末端序列：
SEQ ID NO：18：MF(P)4NDKW没有前导或末端序列：
SEQ ID NO：19：MF(P)4NGWN没有前导或末端序列：
SEQ ID NO：20：NF5PNAIY有前导和末端序列：
SEQ ID NO：21：MF4GC有前导和末端序列：
SEQ ID NO：22：MF4G有前导和末端序列：
SEQ ID NO：23：MF(P)4G有前导和末端序列：
SEQ ID NO：24：MF(P)4NDKW有前导和末端序列：
SEQ ID NO：25：MF(P)4NGWN有前导和末端序列：
SEQ ID NO：26：1/3FLV N-末端
SEQ ID NO：27 2/3FLVN-末端
SEQ ID NO：28 2/3MLV C-末端
SEQ ID NO：29 1/3MLV C-末端
SEQ ID NO：30 FM1(1/3FLV-2/3MLV RT)
SEQ ID NO：31 FM2(2/3FLV-1/3MLV RT)
SEQ ID NO：32野生型MLV RT序列
SEQ ID NO：33野生型FLV RT序列
SEQ ID NO：34：NF5PNAIYC没有前导或末端序列：
SEQ ID NO：35：MF4(P)GC没有前导或末端序列：
SEQ ID NO：36：NF5PNAIYC MLVRT部分：
SEQ ID NO：37：NF5PNAIYC FLVRT部分：
SEQ ID NO：38：MF4(P)GC MLVRT部分
SEQ ID NO：39：MF4(P)GC FLVRT部分
SEQ ID NO：40：NF5PNAIYC有前导和末端序列
SEQ ID NO：41：MF4(P)GC有前导和末端序列
序列表
<110> 生物辐射实验室股份有限公司（Bio-Rad Laboratories, Inc.）
<120> 新型逆转录酶及其用途
<130> 094260-1076461-112710PC
<150> US 62/459,974
<151> 2017-02-16
<160> 42
<170> PatentIn version 3.5
<210> 1
<211> 222
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<220>
<221> MISC_FEATURE
<222> (140)..(140)
<223> Xaa = Leu或Pro
<220>
<221> MISC_FEATURE
<222> (201)..(201)
<223> Xaa = Asp或Asn
<400> 1
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Xaa Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Xaa Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln
210 215 220
<210> 2
<211> 278
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<220>
<221> MISC_FEATURE
<222> (140)..(140)
<223> Xaa = Leu或Pro
<220>
<221> MISC_FEATURE
<222> (201)..(201)
<223> Xaa = Asp或Asn
<400> 2
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Xaa Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Xaa Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln
275
<210> 3
<211> 222
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 3
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln
210 215 220
<210> 4
<211> 278
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 4
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln
275
<210> 5
<211> 391
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<220>
<221> MISC_FEATURE
<222> (245)..(245)
<223> Xaa = Asp, Gly, Asn, 或Ala
<220>
<221> MISC_FEATURE
<222> (249)..(249)
<223> Xaa = Phe或Ile
<220>
<221> MISC_FEATURE
<222> (315)..(315)
<223> Xaa = His或Lys
<220>
<221> MISC_FEATURE
<222> (324)..(324)
<223> Xaa = Leu或Trp
<220>
<221> MISC_FEATURE
<222> (328)..(328)
<223> Xaa = Glu或Lys
<220>
<221> MISC_FEATURE
<222> (355)..(355)
<223> Xaa = His或Tyr
<400> 5
Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu Ser Ile Pro Val
1 5 10 15
Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr
20 25 30
Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr
35 40 45
Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln
50 55 60
Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu
65 70 75 80
Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn
85 90 95
Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu Gly Pro Trp Lys
100 105 110
Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr Val Ala Ser Gly
115 120 125
Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala Ile Leu Val Lys
130 135 140
Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser
145 150 155 160
His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn Lys Trp Leu Ser
165 170 175
Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg
180 185 190
Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala Thr Leu Leu Pro
195 200 205
Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr
210 215 220
Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu
225 230 235 240
Thr Trp Tyr Thr Xaa Gly Ser Ser Xaa Ile Arg Asn Gly Glu Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile Trp Ala Ala Ser
260 265 270
Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Val Xaa Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Xaa Leu Thr Ser Xaa Gly Lys Glu Ile Lys Asn Lys Asn
325 330 335
Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile Xaa Cys Pro Gly His Gln Lys Gly Asp Ser Pro Gln Ala Lys
355 360 365
Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr
370 375 380
Gln Ser Ser Leu Thr Ile Leu
385 390
<210> 6
<211> 406
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<220>
<221> MISC_FEATURE
<222> (245)..(245)
<223> Xaa = Asp, Gly, Asn, 或Ala
<220>
<221> MISC_FEATURE
<222> (249)..(249)
<223> Xaa = Phe或Ile
<220>
<221> MISC_FEATURE
<222> (315)..(315)
<223> Xaa = His或Lys
<220>
<221> MISC_FEATURE
<222> (324)..(324)
<223> Xaa = Leu或Trp
<220>
<221> MISC_FEATURE
<222> (328)..(328)
<223> Xaa = Glu或Lys
<220>
<221> MISC_FEATURE
<222> (355)..(355)
<223> Xaa = His或Tyr
<400> 6
Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu Ser Ile Pro Val
1 5 10 15
Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr
20 25 30
Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr
35 40 45
Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln
50 55 60
Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu
65 70 75 80
Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn
85 90 95
Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu Gly Pro Trp Lys
100 105 110
Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr Val Ala Ser Gly
115 120 125
Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala Ile Leu Val Lys
130 135 140
Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser
145 150 155 160
His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn Lys Trp Leu Ser
165 170 175
Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg
180 185 190
Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala Thr Leu Leu Pro
195 200 205
Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr
210 215 220
Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu
225 230 235 240
Thr Trp Tyr Thr Xaa Gly Ser Ser Xaa Ile Arg Asn Gly Glu Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile Trp Ala Ala Ser
260 265 270
Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Val Xaa Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Xaa Leu Thr Ser Xaa Gly Lys Glu Ile Lys Asn Lys Asn
325 330 335
Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile Xaa Cys Pro Gly His Gln Lys Gly Asp Ser Pro Gln Ala Lys
355 360 365
Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr
370 375 380
Gln Ser Ser Leu Thr Ile Leu Pro Thr Glu Leu Ile Glu Gly Pro Lys
385 390 395 400
Arg Pro Pro Trp Glu Tyr
405
<210> 7
<211> 391
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 7
Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu Ser Ile Pro Val
1 5 10 15
Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr
20 25 30
Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr
35 40 45
Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln
50 55 60
Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu
65 70 75 80
Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn
85 90 95
Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu Gly Pro Trp Lys
100 105 110
Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr Val Ala Ser Gly
115 120 125
Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala Ile Leu Val Lys
130 135 140
Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser
145 150 155 160
His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn Lys Trp Leu Ser
165 170 175
Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg
180 185 190
Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala Thr Leu Leu Pro
195 200 205
Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr
210 215 220
Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu
225 230 235 240
Thr Trp Tyr Thr Ala Gly Ser Ser Ile Ile Arg Asn Gly Glu Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile Trp Ala Ala Ser
260 265 270
Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Val His Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys Glu Ile Lys Asn Lys Asn
325 330 335
Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile Tyr Cys Pro Gly His Gln Lys Gly Asp Ser Pro Gln Ala Lys
355 360 365
Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr
370 375 380
Gln Ser Ser Leu Thr Ile Leu
385 390
<210> 8
<211> 406
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 8
Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu Ser Ile Pro Val
1 5 10 15
Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr
20 25 30
Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr
35 40 45
Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln
50 55 60
Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu
65 70 75 80
Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn
85 90 95
Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu Gly Pro Trp Lys
100 105 110
Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr Val Ala Ser Gly
115 120 125
Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala Ile Leu Val Lys
130 135 140
Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser
145 150 155 160
His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn Lys Trp Leu Ser
165 170 175
Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg
180 185 190
Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala Thr Leu Leu Pro
195 200 205
Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr
210 215 220
Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu
225 230 235 240
Thr Trp Tyr Thr Gly Gly Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile Trp Ala Ala Ser
260 265 270
Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Val His Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Trp Leu Thr Ser Lys Gly Lys Glu Ile Lys Asn Lys Asn
325 330 335
Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile His Cys Pro Gly His Gln Lys Gly Asp Ser Pro Gln Ala Lys
355 360 365
Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr
370 375 380
Gln Ser Ser Leu Thr Ile Leu Pro Thr Glu Leu Ile Glu Gly Pro Lys
385 390 395 400
Arg Pro Pro Trp Glu Tyr
405
<210> 9
<211> 391
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 9
Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu Ser Ile Pro Val
1 5 10 15
Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr
20 25 30
Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr
35 40 45
Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln
50 55 60
Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu
65 70 75 80
Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn
85 90 95
Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu Gly Pro Trp Lys
100 105 110
Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr Val Ala Ser Gly
115 120 125
Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala Ile Leu Val Lys
130 135 140
Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser
145 150 155 160
His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn Lys Trp Leu Ser
165 170 175
Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg
180 185 190
Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala Thr Leu Leu Pro
195 200 205
Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr
210 215 220
Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu
225 230 235 240
Thr Trp Tyr Thr Gly Gly Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile Trp Ala Ala Ser
260 265 270
Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Val His Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Trp Leu Thr Ser Lys Gly Lys Glu Ile Lys Asn Lys Asn
325 330 335
Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile His Cys Pro Gly His Gln Lys Gly Asp Ser Pro Gln Ala Lys
355 360 365
Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr
370 375 380
Gln Ser Ser Leu Thr Ile Leu
385 390
<210> 10
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<220>
<221> MISC_FEATURE
<222> (301)..(301)
<223> Xaa = Asp, Gly, Asn, 或Ala
<220>
<221> MISC_FEATURE
<222> (305)..(305)
<223> Xaa = Phe或Ile
<220>
<221> MISC_FEATURE
<222> (371)..(371)
<223> Xaa = His或Lys
<220>
<221> MISC_FEATURE
<222> (380)..(380)
<223> Xaa = Leu或Trp
<220>
<221> MISC_FEATURE
<222> (384)..(384)
<223> Xaa = Glu或Lys
<220>
<221> MISC_FEATURE
<222> (411)..(411)
<223> Xaa = His或Tyr
<400> 10
Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu
1 5 10 15
Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg
20 25 30
Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu
35 40 45
Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys
50 55 60
Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg
65 70 75 80
Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe
85 90 95
Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu
100 105 110
Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys
115 120 125
Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro
130 135 140
Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu
145 150 155 160
Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys
165 170 175
Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val
180 185 190
Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly
195 200 205
Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg
210 215 220
Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln
225 230 235 240
Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser
245 250 255
Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg
260 265 270
Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp
275 280 285
Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Xaa Gly Ser Ser
290 295 300
Xaa Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu
305 310 315 320
Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln
325 330 335
Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly
340 345 350
Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala
355 360 365
His Val Xaa Gly Glu Ile Tyr Arg Arg Arg Gly Xaa Leu Thr Ser Xaa
370 375 380
Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala
385 390 395 400
Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile Xaa Cys Pro Gly His Gln
405 410 415
Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr
420 425 430
Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
435 440 445
<210> 11
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 11
Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu
1 5 10 15
Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg
20 25 30
Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu
35 40 45
Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys
50 55 60
Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg
65 70 75 80
Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe
85 90 95
Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu
100 105 110
Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys
115 120 125
Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro
130 135 140
Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu
145 150 155 160
Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys
165 170 175
Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val
180 185 190
Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly
195 200 205
Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg
210 215 220
Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln
225 230 235 240
Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser
245 250 255
Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg
260 265 270
Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp
275 280 285
Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser
290 295 300
Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu
305 310 315 320
Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln
325 330 335
Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly
340 345 350
Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala
355 360 365
His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Lys
370 375 380
Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala
385 390 395 400
Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln
405 410 415
Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr
420 425 430
Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
435 440 445
<210> 12
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 12
Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu
1 5 10 15
Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg
20 25 30
Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu
35 40 45
Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys
50 55 60
Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg
65 70 75 80
Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe
85 90 95
Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu
100 105 110
Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys
115 120 125
Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro
130 135 140
Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu
145 150 155 160
Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys
165 170 175
Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val
180 185 190
Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly
195 200 205
Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg
210 215 220
Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln
225 230 235 240
Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser
245 250 255
Leu Asp Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg
260 265 270
Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp
275 280 285
Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Asp Gly Ser Ser
290 295 300
Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu
305 310 315 320
Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln
325 330 335
Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly
340 345 350
Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala
355 360 365
His Val Lys Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Glu
370 375 380
Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala
385 390 395 400
Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln
405 410 415
Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr
420 425 430
Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
435 440 445
<210> 13
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 13
Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu
1 5 10 15
Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg
20 25 30
Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu
35 40 45
Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys
50 55 60
Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg
65 70 75 80
Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe
85 90 95
Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu
100 105 110
Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys
115 120 125
Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro
130 135 140
Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu
145 150 155 160
Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys
165 170 175
Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val
180 185 190
Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly
195 200 205
Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg
210 215 220
Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln
225 230 235 240
Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser
245 250 255
Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg
260 265 270
Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp
275 280 285
Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser
290 295 300
Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu
305 310 315 320
Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln
325 330 335
Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly
340 345 350
Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala
355 360 365
His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Glu
370 375 380
Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala
385 390 395 400
Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln
405 410 415
Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asn Asp Thr
420 425 430
Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
435 440 445
<210> 14
<211> 669
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 14
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Ala Gly Ser Ser Ile Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile Tyr Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
660 665
<210> 15
<211> 684
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 15
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser Phe Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Lys Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu Pro Thr Glu
660 665 670
Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
675 680
<210> 16
<211> 669
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 16
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser Phe Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Lys Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
660 665
<210> 17
<211> 669
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 17
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly
225 230 235 240
Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr
260 265 270
Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser Phe Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Lys Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
660 665
<210> 18
<211> 669
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 18
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Leu Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly
225 230 235 240
Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr
260 265 270
Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asp
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Asp Gly Ser Ser Phe Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
Lys Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Glu Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
660 665
<210> 19
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 19
Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu
1 5 10 15
Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg
20 25 30
Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu
35 40 45
Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys
50 55 60
Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg
65 70 75 80
Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe
85 90 95
Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu
100 105 110
Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys
115 120 125
Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro
130 135 140
Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu
145 150 155 160
Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys
165 170 175
Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val
180 185 190
Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly
195 200 205
Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg
210 215 220
Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln
225 230 235 240
Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser
245 250 255
Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg
260 265 270
Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp
275 280 285
Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser
290 295 300
Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu
305 310 315 320
Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln
325 330 335
Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly
340 345 350
Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala
355 360 365
His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Glu
370 375 380
Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala
385 390 395 400
Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln
405 410 415
Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asn Asp Thr
420 425 430
Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
435 440 445
<210> 20
<211> 692
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 20
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp
245 250 255
Cys Gln Gln Gly Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys
275 280 285
Tyr Leu Gly Tyr Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Ala Gly
530 535 540
Ser Ser Ile Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr
610 615 620
Ser Glu Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile Tyr Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Ala Ala Ala
690
<210> 21
<211> 707
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 21
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp
245 250 255
Cys Gln Gln Gly Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys
275 280 285
Tyr Leu Gly Tyr Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly
530 535 540
Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr
610 615 620
Ser Lys Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Pro Thr Glu Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
690 695 700
Ala Ala Ala
705
<210> 22
<211> 692
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 22
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp
245 250 255
Cys Gln Gln Gly Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys
275 280 285
Tyr Leu Gly Tyr Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly
530 535 540
Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr
610 615 620
Ser Lys Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Ala Ala Ala
690
<210> 23
<211> 692
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 23
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu
245 250 255
Cys Leu Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr
275 280 285
Tyr Leu Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly
530 535 540
Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr
610 615 620
Ser Lys Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Ala Ala Ala
690
<210> 24
<211> 692
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 24
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Leu
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu
245 250 255
Cys Leu Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr
275 280 285
Tyr Leu Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asp Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Asp Gly
530 535 540
Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val Lys Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr
610 615 620
Ser Glu Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Ala Ala Ala
690
<210> 25
<211> 692
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 25
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Leu
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu
245 250 255
Cys Leu Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr
275 280 285
Tyr Leu Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly
530 535 540
Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr
610 615 620
Ser Glu Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asn
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Ala Ala Ala
690
<210> 26
<211> 279
<212> PRT
<213> 未知
<220>
<223> 猫白血病病毒
<400> 26
Met Thr Leu Gln Leu Glu Glu Glu Tyr Arg Leu Phe Glu Pro Glu Ser
1 5 10 15
Thr Gln Lys Gln Glu Met Asp Ile Trp Leu Lys Asn Phe Pro Gln Ala
20 25 30
Trp Ala Glu Thr Gly Gly Met Gly Met Ala His Cys Gln Ala Pro Val
35 40 45
Leu Ile Gln Leu Lys Ala Thr Ala Thr Pro Ile Ser Ile Arg Gln Tyr
50 55 60
Pro Met Pro His Glu Ala Tyr Gln Gly Ile Lys Pro His Ile Arg Arg
65 70 75 80
Met Leu Asp Gln Gly Ile Leu Lys Pro Cys Gln Ser Pro Trp Asn Thr
85 90 95
Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Lys Asp Tyr Arg Pro Val
100 105 110
Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro Thr
115 120 125
Val Pro Asn Pro Tyr Asn Leu Leu Ser Thr Leu Pro Pro Ser His Pro
130 135 140
Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg Leu
145 150 155 160
His Ser Glu Ser Gln Leu Leu Phe Ala Phe Glu Trp Arg Asp Pro Glu
165 170 175
Ile Gly Leu Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly Phe
180 185 190
Lys Asn Ser Pro Thr Leu Phe Asp Glu Ala Leu His Ser Asp Leu Ala
195 200 205
Asp Phe Arg Val Arg Tyr Pro Ala Leu Val Leu Leu Gln Tyr Val Asp
210 215 220
Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly Thr
225 230 235 240
Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser Ala
245 250 255
Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr Ser
260 265 270
Leu Lys Asp Gly Gln Arg Trp
275
<210> 27
<211> 497
<212> PRT
<213> 未知
<220>
<223> 猫白血病病毒
<400> 27
Met Thr Leu Gln Leu Glu Glu Glu Tyr Arg Leu Phe Glu Pro Glu Ser
1 5 10 15
Thr Gln Lys Gln Glu Met Asp Ile Trp Leu Lys Asn Phe Pro Gln Ala
20 25 30
Trp Ala Glu Thr Gly Gly Met Gly Met Ala His Cys Gln Ala Pro Val
35 40 45
Leu Ile Gln Leu Lys Ala Thr Ala Thr Pro Ile Ser Ile Arg Gln Tyr
50 55 60
Pro Met Pro His Glu Ala Tyr Gln Gly Ile Lys Pro His Ile Arg Arg
65 70 75 80
Met Leu Asp Gln Gly Ile Leu Lys Pro Cys Gln Ser Pro Trp Asn Thr
85 90 95
Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Lys Asp Tyr Arg Pro Val
100 105 110
Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro Thr
115 120 125
Val Pro Asn Pro Tyr Asn Leu Leu Ser Thr Leu Pro Pro Ser His Pro
130 135 140
Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg Leu
145 150 155 160
His Ser Glu Ser Gln Leu Leu Phe Ala Phe Glu Trp Arg Asp Pro Glu
165 170 175
Ile Gly Leu Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly Phe
180 185 190
Lys Asn Ser Pro Thr Leu Phe Asp Glu Ala Leu His Ser Asp Leu Ala
195 200 205
Asp Phe Arg Val Arg Tyr Pro Ala Leu Val Leu Leu Gln Tyr Val Asp
210 215 220
Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly Thr
225 230 235 240
Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser Ala
245 250 255
Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr Ser
260 265 270
Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile
275 280 285
Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu
290 295 300
Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu
305 310 315 320
Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp
325 330 335
Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu
340 345 350
Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu
355 360 365
Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys
370 375 380
Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp
385 390 395 400
Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile
405 410 415
Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu
420 425 430
Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro Pro
435 440 445
Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu
450 455 460
Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn Pro
465 470 475 480
Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro
485 490 495
Asp
<210> 28
<211> 392
<212> PRT
<213> 未知
<220>
<223> 小鼠白血病病毒
<400> 28
Leu Thr Glu Ala Arg Lys Glu Thr Val Met Gly Gln Pro Thr Pro Lys
1 5 10 15
Thr Pro Arg Gln Leu Arg Glu Phe Leu Gly Thr Ala Gly Phe Cys Arg
20 25 30
Leu Trp Ile Pro Gly Phe Ala Glu Met Ala Ala Pro Leu Tyr Pro Leu
35 40 45
Thr Lys Thr Gly Thr Leu Phe Asn Trp Gly Pro Asp Gln Gln Lys Ala
50 55 60
Tyr Gln Glu Ile Lys Gln Ala Leu Leu Thr Ala Pro Ala Leu Gly Leu
65 70 75 80
Pro Asp Leu Thr Lys Pro Phe Glu Leu Phe Val Asp Glu Lys Gln Gly
85 90 95
Tyr Ala Lys Gly Val Leu Thr Gln Lys Leu Gly Pro Trp Arg Arg Pro
100 105 110
Val Ala Tyr Leu Ser Lys Lys Leu Asp Pro Val Ala Ala Gly Trp Pro
115 120 125
Pro Cys Leu Arg Met Val Ala Ala Ile Ala Val Leu Thr Lys Asp Ala
130 135 140
Gly Lys Leu Thr Met Gly Gln Pro Leu Val Ile Leu Ala Pro His Ala
145 150 155 160
Val Glu Ala Leu Val Lys Gln Pro Pro Asp Arg Trp Leu Ser Asn Ala
165 170 175
Arg Met Thr His Tyr Gln Ala Leu Leu Leu Asp Thr Asp Arg Val Gln
180 185 190
Phe Gly Pro Val Val Ala Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro
195 200 205
Glu Glu Gly Leu Gln His Asp Cys Leu Asp Ile Leu Ala Glu Ala His
210 215 220
Gly Thr Arg Ser Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp His
225 230 235 240
Thr Trp Tyr Thr Asp Gly Ser Ser Phe Leu Gln Glu Gly Gln Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Thr Glu Val Ile Trp Ala Arg Ala
260 265 270
Leu Pro Ala Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Glu Gly Lys Lys Leu Asn Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Ile His Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys Glu Ile Lys Asn Lys Asp
325 330 335
Glu Ile Leu Ala Leu Leu Lys Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile His Cys Pro Gly His Gln Lys Gly Asn Ser Ala Glu Ala Arg
355 360 365
Gly Asn Arg Met Ala Asp Gln Ala Ala Arg Glu Val Ala Thr Arg Glu
370 375 380
Thr Pro Gly Thr Ser Thr Leu Leu
385 390
<210> 29
<211> 174
<212> PRT
<213> 未知
<220>
<223> 小鼠白血病病毒
<400> 29
Ile Leu Ala Glu Ala His Gly Thr Arg Ser Asp Leu Thr Asp Gln Pro
1 5 10 15
Leu Pro Asp Ala Asp His Thr Trp Tyr Thr Asp Gly Ser Ser Phe Leu
20 25 30
Gln Glu Gly Gln Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Thr Glu
35 40 45
Val Ile Trp Ala Arg Ala Leu Pro Ala Gly Thr Ser Ala Gln Arg Ala
50 55 60
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Glu Gly Lys Lys
65 70 75 80
Leu Asn Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Ile
85 90 95
His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys
100 105 110
Glu Ile Lys Asn Lys Asp Glu Ile Leu Ala Leu Leu Lys Ala Leu Phe
115 120 125
Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly
130 135 140
Asn Ser Ala Glu Ala Arg Gly Asn Arg Met Ala Asp Gln Ala Ala Arg
145 150 155 160
Glu Val Ala Thr Arg Glu Thr Pro Gly Thr Ser Thr Leu Leu
165 170
<210> 30
<211> 671
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 30
Met Thr Leu Gln Leu Glu Glu Glu Tyr Arg Leu Phe Glu Pro Glu Ser
1 5 10 15
Thr Gln Lys Gln Glu Met Asp Ile Trp Leu Lys Asn Phe Pro Gln Ala
20 25 30
Trp Ala Glu Thr Gly Gly Met Gly Met Ala His Cys Gln Ala Pro Val
35 40 45
Leu Ile Gln Leu Lys Ala Thr Ala Thr Pro Ile Ser Ile Arg Gln Tyr
50 55 60
Pro Met Pro His Glu Ala Tyr Gln Gly Ile Lys Pro His Ile Arg Arg
65 70 75 80
Met Leu Asp Gln Gly Ile Leu Lys Pro Cys Gln Ser Pro Trp Asn Thr
85 90 95
Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Lys Asp Tyr Arg Pro Val
100 105 110
Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro Thr
115 120 125
Val Pro Asn Pro Tyr Asn Leu Leu Ser Thr Leu Pro Pro Ser His Pro
130 135 140
Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg Leu
145 150 155 160
His Ser Glu Ser Gln Leu Leu Phe Ala Phe Glu Trp Arg Asp Pro Glu
165 170 175
Ile Gly Leu Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly Phe
180 185 190
Lys Asn Ser Pro Thr Leu Phe Asp Glu Ala Leu His Ser Asp Leu Ala
195 200 205
Asp Phe Arg Val Arg Tyr Pro Ala Leu Val Leu Leu Gln Tyr Val Asp
210 215 220
Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly Thr
225 230 235 240
Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser Ala
245 250 255
Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr Ser
260 265 270
Leu Lys Asp Gly Gln Arg Trp Leu Thr Glu Ala Arg Lys Glu Thr Val
275 280 285
Met Gly Gln Pro Thr Pro Lys Thr Pro Arg Gln Leu Arg Glu Phe Leu
290 295 300
Gly Thr Ala Gly Phe Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Met
305 310 315 320
Ala Ala Pro Leu Tyr Pro Leu Thr Lys Thr Gly Thr Leu Phe Asn Trp
325 330 335
Gly Pro Asp Gln Gln Lys Ala Tyr Gln Glu Ile Lys Gln Ala Leu Leu
340 345 350
Thr Ala Pro Ala Leu Gly Leu Pro Asp Leu Thr Lys Pro Phe Glu Leu
355 360 365
Phe Val Asp Glu Lys Gln Gly Tyr Ala Lys Gly Val Leu Thr Gln Lys
370 375 380
Leu Gly Pro Trp Arg Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp
385 390 395 400
Pro Val Ala Ala Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile
405 410 415
Ala Val Leu Thr Lys Asp Ala Gly Lys Leu Thr Met Gly Gln Pro Leu
420 425 430
Val Ile Leu Ala Pro His Ala Val Glu Ala Leu Val Lys Gln Pro Pro
435 440 445
Asp Arg Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Leu Leu
450 455 460
Leu Asp Thr Asp Arg Val Gln Phe Gly Pro Val Val Ala Leu Asn Pro
465 470 475 480
Ala Thr Leu Leu Pro Leu Pro Glu Glu Gly Leu Gln His Asp Cys Leu
485 490 495
Asp Ile Leu Ala Glu Ala His Gly Thr Arg Ser Asp Leu Thr Asp Gln
500 505 510
Pro Leu Pro Asp Ala Asp His Thr Trp Tyr Thr Asp Gly Ser Ser Phe
515 520 525
Leu Gln Glu Gly Gln Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Thr
530 535 540
Glu Val Ile Trp Ala Arg Ala Leu Pro Ala Gly Thr Ser Ala Gln Arg
545 550 555 560
Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Glu Gly Lys
565 570 575
Lys Leu Asn Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His
580 585 590
Ile His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly
595 600 605
Lys Glu Ile Lys Asn Lys Asp Glu Ile Leu Ala Leu Leu Lys Ala Leu
610 615 620
Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys
625 630 635 640
Gly Asn Ser Ala Glu Ala Arg Gly Asn Arg Met Ala Asp Gln Ala Ala
645 650 655
Arg Glu Val Ala Thr Arg Glu Thr Pro Gly Thr Ser Thr Leu Leu
660 665 670
<210> 31
<211> 671
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 31
Met Thr Leu Gln Leu Glu Glu Glu Tyr Arg Leu Phe Glu Pro Glu Ser
1 5 10 15
Thr Gln Lys Gln Glu Met Asp Ile Trp Leu Lys Asn Phe Pro Gln Ala
20 25 30
Trp Ala Glu Thr Gly Gly Met Gly Met Ala His Cys Gln Ala Pro Val
35 40 45
Leu Ile Gln Leu Lys Ala Thr Ala Thr Pro Ile Ser Ile Arg Gln Tyr
50 55 60
Pro Met Pro His Glu Ala Tyr Gln Gly Ile Lys Pro His Ile Arg Arg
65 70 75 80
Met Leu Asp Gln Gly Ile Leu Lys Pro Cys Gln Ser Pro Trp Asn Thr
85 90 95
Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Lys Asp Tyr Arg Pro Val
100 105 110
Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro Thr
115 120 125
Val Pro Asn Pro Tyr Asn Leu Leu Ser Thr Leu Pro Pro Ser His Pro
130 135 140
Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg Leu
145 150 155 160
His Ser Glu Ser Gln Leu Leu Phe Ala Phe Glu Trp Arg Asp Pro Glu
165 170 175
Ile Gly Leu Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly Phe
180 185 190
Lys Asn Ser Pro Thr Leu Phe Asp Glu Ala Leu His Ser Asp Leu Ala
195 200 205
Asp Phe Arg Val Arg Tyr Pro Ala Leu Val Leu Leu Gln Tyr Val Asp
210 215 220
Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly Thr
225 230 235 240
Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser Ala
245 250 255
Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr Ser
260 265 270
Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile
275 280 285
Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu
290 295 300
Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu
305 310 315 320
Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp
325 330 335
Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu
340 345 350
Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu
355 360 365
Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys
370 375 380
Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp
385 390 395 400
Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile
405 410 415
Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu
420 425 430
Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro Pro
435 440 445
Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu
450 455 460
Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn Pro
465 470 475 480
Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro
485 490 495
Asp Ile Leu Ala Glu Ala His Gly Thr Arg Ser Asp Leu Thr Asp Gln
500 505 510
Pro Leu Pro Asp Ala Asp His Thr Trp Tyr Thr Asp Gly Ser Ser Phe
515 520 525
Leu Gln Glu Gly Gln Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Thr
530 535 540
Glu Val Ile Trp Ala Arg Ala Leu Pro Ala Gly Thr Ser Ala Gln Arg
545 550 555 560
Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Glu Gly Lys
565 570 575
Lys Leu Asn Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His
580 585 590
Ile His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly
595 600 605
Lys Glu Ile Lys Asn Lys Asp Glu Ile Leu Ala Leu Leu Lys Ala Leu
610 615 620
Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys
625 630 635 640
Gly Asn Ser Ala Glu Ala Arg Gly Asn Arg Met Ala Asp Gln Ala Ala
645 650 655
Arg Glu Val Ala Thr Arg Glu Thr Pro Gly Thr Ser Thr Leu Leu
660 665 670
<210> 32
<211> 671
<212> PRT
<213> 未知
<220>
<223> 小鼠白血病病毒
<400> 32
Thr Leu Asn Ile Glu Asp Glu His Arg Leu His Glu Thr Ser Lys Glu
1 5 10 15
Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln Ala
20 25 30
Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro Leu
35 40 45
Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln Tyr
50 55 60
Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln Arg
65 70 75 80
Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn Thr
85 90 95
Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro Val
100 105 110
Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro Thr
115 120 125
Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Leu Pro Pro Ser His Gln
130 135 140
Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg Leu
145 150 155 160
His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro Glu
165 170 175
Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly Phe
180 185 190
Lys Asn Ser Pro Thr Leu Phe Asp Glu Ala Leu His Arg Asp Leu Ala
195 200 205
Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val Asp
210 215 220
Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly Thr
225 230 235 240
Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser Ala
245 250 255
Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr Leu
260 265 270
Leu Lys Glu Gly Gln Arg Trp Leu Thr Glu Ala Arg Lys Glu Thr Val
275 280 285
Met Gly Gln Pro Thr Pro Lys Thr Pro Arg Gln Leu Arg Glu Phe Leu
290 295 300
Gly Thr Ala Gly Phe Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Met
305 310 315 320
Ala Ala Pro Leu Tyr Pro Leu Thr Lys Thr Gly Thr Leu Phe Asn Trp
325 330 335
Gly Pro Asp Gln Gln Lys Ala Tyr Gln Glu Ile Lys Gln Ala Leu Leu
340 345 350
Thr Ala Pro Ala Leu Gly Leu Pro Asp Leu Thr Lys Pro Phe Glu Leu
355 360 365
Phe Val Asp Glu Lys Gln Gly Tyr Ala Lys Gly Val Leu Thr Gln Lys
370 375 380
Leu Gly Pro Trp Arg Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp
385 390 395 400
Pro Val Ala Ala Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile
405 410 415
Ala Val Leu Thr Lys Asp Ala Gly Lys Leu Thr Met Gly Gln Pro Leu
420 425 430
Val Ile Leu Ala Pro His Ala Val Glu Ala Leu Val Lys Gln Pro Pro
435 440 445
Asp Arg Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Leu Leu
450 455 460
Leu Asp Thr Asp Arg Val Gln Phe Gly Pro Val Val Ala Leu Asn Pro
465 470 475 480
Ala Thr Leu Leu Pro Leu Pro Glu Glu Gly Leu Gln His Asn Cys Leu
485 490 495
Asp Ile Leu Ala Glu Ala His Gly Thr Arg Pro Asp Leu Thr Asp Gln
500 505 510
Pro Leu Pro Asp Ala Asp His Thr Trp Tyr Thr Asp Gly Ser Ser Leu
515 520 525
Leu Gln Glu Gly Gln Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Thr
530 535 540
Glu Val Ile Trp Ala Lys Ala Leu Pro Ala Gly Thr Ser Ala Gln Arg
545 550 555 560
Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Glu Gly Lys
565 570 575
Lys Leu Asn Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His
580 585 590
Ile His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly
595 600 605
Lys Glu Ile Lys Asn Lys Asp Glu Ile Leu Ala Leu Leu Lys Ala Leu
610 615 620
Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys
625 630 635 640
Gly His Ser Ala Glu Ala Arg Gly Asn Arg Met Ala Asp Gln Ala Ala
645 650 655
Arg Lys Ala Ala Ile Thr Glu Thr Pro Asp Thr Ser Thr Leu Leu
660 665 670
<210> 33
<211> 667
<212> PRT
<213> 未知
<220>
<223> 猫白血病病毒
<400> 33
Thr Leu Gln Leu Glu Glu Glu Tyr Arg Leu Phe Glu Pro Glu Ser Thr
1 5 10 15
Gln Lys Gln Glu Met Asp Ile Trp Leu Lys Asn Phe Pro Gln Ala Trp
20 25 30
Ala Glu Thr Gly Gly Met Gly Met Ala His Cys Gln Ala Pro Val Leu
35 40 45
Ile Gln Leu Lys Ala Thr Ala Thr Pro Ile Ser Ile Arg Gln Tyr Pro
50 55 60
Met Pro His Glu Ala Tyr Gln Gly Ile Lys Pro His Ile Arg Arg Met
65 70 75 80
Leu Asp Gln Gly Ile Leu Lys Pro Cys Gln Ser Pro Trp Asn Thr Pro
85 90 95
Leu Leu Pro Val Lys Lys Pro Gly Thr Lys Asp Tyr Arg Pro Val Gln
100 105 110
Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro Thr Val
115 120 125
Pro Asn Pro Tyr Asn Leu Leu Ser Thr Leu Pro Pro Ser His Pro Trp
130 135 140
Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg Leu His
145 150 155 160
Ser Glu Ser Gln Leu Leu Phe Ala Phe Glu Trp Arg Asp Pro Glu Ile
165 170 175
Gly Leu Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly Phe Lys
180 185 190
Asn Ser Pro Thr Leu Phe Asp Glu Ala Leu His Ser Asp Leu Ala Asp
195 200 205
Phe Arg Val Arg Tyr Pro Ala Leu Val Leu Leu Gln Tyr Val Asp Asp
210 215 220
Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly Thr Lys
225 230 235 240
Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser Ala Lys
245 250 255
Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr Ser Leu
260 265 270
Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu
275 280 285
Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly
290 295 300
Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala
305 310 315 320
Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly
325 330 335
Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser
340 345 350
Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe
355 360 365
Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu
370 375 380
Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr
385 390 395 400
Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala
405 410 415
Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr
420 425 430
Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn
435 440 445
Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu
450 455 460
Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala
465 470 475 480
Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp
485 490 495
Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro
500 505 510
Asp Ala Asp Leu Thr Trp Tyr Thr Asp Gly Ser Ser Phe Ile Arg Asn
515 520 525
Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile
530 535 540
Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu
545 550 555 560
Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr
565 570 575
Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val His Gly
580 585 590
Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys Glu Ile
595 600 605
Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro
610 615 620
Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly Asp Ser
625 630 635 640
Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala
645 650 655
Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu
660 665
<210> 34
<211> 684
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 34
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Ala Gly Ser Ser Ile Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile Tyr Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu Pro Thr Glu
660 665 670
Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
675 680
<210> 35
<211> 684
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 35
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu Glu Gly
225 230 235 240
Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu Gly Tyr
260 265 270
Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala
275 280 285
Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg Glu Phe
290 295 300
Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu
305 310 315 320
Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln
325 330 335
Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu
340 345 350
Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu
355 360 365
Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu Val Gln
370 375 380
Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu
385 390 395 400
Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val Ala Ala
405 410 415
Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro
420 425 430
Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg Gln Pro
435 440 445
Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln Ala Met
450 455 460
Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser Leu Asn
465 470 475 480
Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser
485 490 495
Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp Gln Pro
500 505 510
Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser Phe Ile
515 520 525
Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu Ser Glu
530 535 540
Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala
545 550 555 560
Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly Lys Lys
565 570 575
Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala His Val
580 585 590
His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Lys Gly Lys
595 600 605
Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe
610 615 620
Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln Lys Gly
625 630 635 640
Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys
645 650 655
Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu Pro Thr Glu
660 665 670
Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
675 680
<210> 36
<211> 278
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 36
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln Tyr Val
210 215 220
Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp Cys Gln Gln Gly
225 230 235 240
Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly Tyr Arg Ala Ser
245 250 255
Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys Tyr Leu Gly Tyr
260 265 270
Leu Leu Lys Glu Gly Gln
275
<210> 37
<211> 406
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 37
Arg Trp Leu Thr Lys Ala Arg Lys Glu Ala Ile Leu Ser Ile Pro Val
1 5 10 15
Pro Lys Asn Pro Arg Gln Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr
20 25 30
Cys Arg Leu Trp Ile Pro Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr
35 40 45
Pro Leu Thr Arg Pro Gly Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln
50 55 60
Leu Ala Phe Glu Asn Ile Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu
65 70 75 80
Gly Leu Pro Asp Ile Thr Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn
85 90 95
Ser Gly Phe Ala Lys Gly Val Leu Val Gln Lys Leu Gly Pro Trp Lys
100 105 110
Arg Pro Val Ala Tyr Leu Ser Lys Lys Leu Asp Thr Val Ala Ser Gly
115 120 125
Trp Pro Pro Cys Leu Arg Met Val Ala Ala Ile Ala Ile Leu Val Lys
130 135 140
Asp Ala Gly Lys Leu Thr Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser
145 150 155 160
His Pro Val Glu Ala Leu Val Arg Gln Pro Pro Asn Lys Trp Leu Ser
165 170 175
Asn Ala Arg Met Thr His Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg
180 185 190
Val His Phe Gly Pro Thr Val Ser Leu Asn Pro Ala Thr Leu Leu Pro
195 200 205
Leu Pro Ser Gly Lys Pro Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr
210 215 220
Met Ala Gln Thr Asp Leu Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu
225 230 235 240
Thr Trp Tyr Thr Ala Gly Ser Ser Ile Ile Arg Asn Gly Glu Arg Lys
245 250 255
Ala Gly Ala Ala Val Thr Thr Glu Ser Glu Val Ile Trp Ala Ala Ser
260 265 270
Leu Pro Pro Gly Thr Ser Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr
275 280 285
Gln Ala Leu Lys Met Ala Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp
290 295 300
Ser Arg Tyr Ala Phe Ala Thr Ala His Val His Gly Glu Ile Tyr Arg
305 310 315 320
Arg Arg Gly Leu Leu Thr Ser Glu Gly Lys Glu Ile Lys Asn Lys Asn
325 330 335
Glu Ile Leu Ala Leu Leu Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser
340 345 350
Ile Ile Tyr Cys Pro Gly His Gln Lys Gly Asp Ser Pro Gln Ala Lys
355 360 365
Gly Asn Arg Leu Ala Asp Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr
370 375 380
Gln Ser Ser Leu Thr Ile Leu Pro Thr Glu Leu Ile Glu Gly Pro Lys
385 390 395 400
Arg Pro Pro Trp Glu Tyr
405
<210> 38
<211> 222
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 38
Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His Glu Thr Ser Lys
1 5 10 15
Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser Asp Phe Pro Gln
20 25 30
Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val Arg Gln Ala Pro
35 40 45
Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val Ser Ile Lys Gln
50 55 60
Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys Pro His Ile Gln
65 70 75 80
Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln Ser Pro Trp Asn
85 90 95
Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn Asp Tyr Arg Pro
100 105 110
Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu Asp Ile His Pro
115 120 125
Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro Pro Pro Ser His
130 135 140
Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe Phe Cys Leu Arg
145 150 155 160
Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu Trp Arg Asp Pro
165 170 175
Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg Leu Pro Gln Gly
180 185 190
Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu His Arg Asp Leu
195 200 205
Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu Leu Gln
210 215 220
<210> 39
<211> 462
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 39
Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu Cys Leu
1 5 10 15
Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly Tyr Arg
20 25 30
Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr Tyr Leu
35 40 45
Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala Arg Lys
50 55 60
Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln Val Arg
65 70 75 80
Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro Gly Phe
85 90 95
Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly Thr Leu
100 105 110
Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile Arg Lys
115 120 125
Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr Lys Pro
130 135 140
Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly Val Leu
145 150 155 160
Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu Ser Lys
165 170 175
Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg Met Val
180 185 190
Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr Leu Gly
195 200 205
Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu Val Arg
210 215 220
Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His Tyr Gln
225 230 235 240
Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr Val Ser
245 250 255
Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro Pro Arg
260 265 270
Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu Thr Asp
275 280 285
Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly Ser Ser
290 295 300
Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr Thr Glu
305 310 315 320
Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser Ala Gln
325 330 335
Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala Lys Gly
340 345 350
Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala Thr Ala
355 360 365
His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr Ser Lys
370 375 380
Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu Glu Ala
385 390 395 400
Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly His Gln
405 410 415
Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp Asp Thr
420 425 430
Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile Leu Pro
435 440 445
Thr Glu Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
450 455 460
<210> 40
<211> 707
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 40
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Thr Ser Glu Leu Asp
245 250 255
Cys Gln Gln Gly Thr Arg Ala Leu Leu Gln Thr Leu Gly Asn Leu Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Gln Lys Gln Val Lys
275 280 285
Tyr Leu Gly Tyr Leu Leu Lys Glu Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Ala Gly
530 535 540
Ser Ser Ile Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Leu Leu Thr
610 615 620
Ser Glu Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile Tyr Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Pro Thr Glu Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
690 695 700
Ala Ala Ala
705
<210> 41
<211> 707
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 41
Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro
1 5 10 15
Arg Gly Ser His Met Thr Leu Asn Ile Glu Asp Glu Tyr Arg Leu His
20 25 30
Glu Thr Ser Lys Glu Pro Asp Val Ser Leu Gly Ser Thr Trp Leu Ser
35 40 45
Asp Phe Pro Gln Ala Trp Ala Glu Thr Gly Gly Met Gly Leu Ala Val
50 55 60
Arg Gln Ala Pro Leu Ile Ile Pro Leu Lys Ala Thr Ser Thr Pro Val
65 70 75 80
Ser Ile Lys Gln Tyr Pro Met Ser Gln Glu Ala Arg Leu Gly Ile Lys
85 90 95
Pro His Ile Gln Arg Leu Leu Asp Gln Gly Ile Leu Val Pro Cys Gln
100 105 110
Ser Pro Trp Asn Thr Pro Leu Leu Pro Val Lys Lys Pro Gly Thr Asn
115 120 125
Asp Tyr Arg Pro Val Gln Asp Leu Arg Glu Val Asn Lys Arg Val Glu
130 135 140
Asp Ile His Pro Thr Val Pro Asn Pro Tyr Asn Leu Leu Ser Gly Pro
145 150 155 160
Pro Pro Ser His Gln Trp Tyr Thr Val Leu Asp Leu Lys Asp Ala Phe
165 170 175
Phe Cys Leu Arg Leu His Pro Thr Ser Gln Pro Leu Phe Ala Phe Glu
180 185 190
Trp Arg Asp Pro Glu Met Gly Ile Ser Gly Gln Leu Thr Trp Thr Arg
195 200 205
Leu Pro Gln Gly Phe Lys Asn Ser Pro Thr Leu Phe Asn Glu Ala Leu
210 215 220
His Arg Asp Leu Ala Asp Phe Arg Ile Gln His Pro Asp Leu Ile Leu
225 230 235 240
Leu Gln Tyr Val Asp Asp Leu Leu Leu Ala Ala Ala Thr Arg Thr Glu
245 250 255
Cys Leu Glu Gly Thr Lys Ala Leu Leu Glu Thr Leu Gly Asn Lys Gly
260 265 270
Tyr Arg Ala Ser Ala Lys Lys Ala Gln Ile Cys Leu Gln Glu Val Thr
275 280 285
Tyr Leu Gly Tyr Ser Leu Lys Asp Gly Gln Arg Trp Leu Thr Lys Ala
290 295 300
Arg Lys Glu Ala Ile Leu Ser Ile Pro Val Pro Lys Asn Pro Arg Gln
305 310 315 320
Val Arg Glu Phe Leu Gly Thr Ala Gly Tyr Cys Arg Leu Trp Ile Pro
325 330 335
Gly Phe Ala Glu Leu Ala Ala Pro Leu Tyr Pro Leu Thr Arg Pro Gly
340 345 350
Thr Leu Phe Gln Trp Gly Thr Glu Gln Gln Leu Ala Phe Glu Asn Ile
355 360 365
Arg Lys Ala Leu Leu Ser Ser Pro Ala Leu Gly Leu Pro Asp Ile Thr
370 375 380
Lys Pro Phe Glu Leu Phe Ile Asp Glu Asn Ser Gly Phe Ala Lys Gly
385 390 395 400
Val Leu Val Gln Lys Leu Gly Pro Trp Lys Arg Pro Val Ala Tyr Leu
405 410 415
Ser Lys Lys Leu Asp Thr Val Ala Ser Gly Trp Pro Pro Cys Leu Arg
420 425 430
Met Val Ala Ala Ile Ala Ile Leu Val Lys Asp Ala Gly Lys Leu Thr
435 440 445
Leu Gly Gln Pro Leu Thr Ile Leu Thr Ser His Pro Val Glu Ala Leu
450 455 460
Val Arg Gln Pro Pro Asn Lys Trp Leu Ser Asn Ala Arg Met Thr His
465 470 475 480
Tyr Gln Ala Met Leu Leu Asp Ala Glu Arg Val His Phe Gly Pro Thr
485 490 495
Val Ser Leu Asn Pro Ala Thr Leu Leu Pro Leu Pro Ser Gly Lys Pro
500 505 510
Pro Arg Leu Ser Pro Asp Leu Ala Glu Thr Met Ala Gln Thr Asp Leu
515 520 525
Thr Asp Gln Pro Leu Pro Asp Ala Asp Leu Thr Trp Tyr Thr Gly Gly
530 535 540
Ser Ser Phe Ile Arg Asn Gly Glu Arg Lys Ala Gly Ala Ala Val Thr
545 550 555 560
Thr Glu Ser Glu Val Ile Trp Ala Ala Ser Leu Pro Pro Gly Thr Ser
565 570 575
Ala Gln Arg Ala Glu Leu Ile Ala Leu Thr Gln Ala Leu Lys Met Ala
580 585 590
Lys Gly Lys Lys Leu Thr Val Tyr Thr Asp Ser Arg Tyr Ala Phe Ala
595 600 605
Thr Ala His Val His Gly Glu Ile Tyr Arg Arg Arg Gly Trp Leu Thr
610 615 620
Ser Lys Gly Lys Glu Ile Lys Asn Lys Asn Glu Ile Leu Ala Leu Leu
625 630 635 640
Glu Ala Leu Phe Leu Pro Lys Arg Leu Ser Ile Ile His Cys Pro Gly
645 650 655
His Gln Lys Gly Asp Ser Pro Gln Ala Lys Gly Asn Arg Leu Ala Asp
660 665 670
Asp Thr Ala Lys Lys Ala Ala Thr Glu Thr Gln Ser Ser Leu Thr Ile
675 680 685
Leu Pro Thr Glu Leu Ile Glu Gly Pro Lys Arg Pro Pro Trp Glu Tyr
690 695 700
Ala Ala Ala
705
<210> 42
<211> 8
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 42
Asp Tyr Lys Asp Asp Asp Asp Lys
1 5

Claims (21)

1.一种杂合逆转录酶，其包含指结构域，掌结构域，拇指结构域，连接结构域和RNA酶H结构域，其中至少一个所述结构域是小鼠白血病病毒逆转录酶(MLVRT)而其他所述结构域来自猫白血病病毒逆转录酶(FLVRT)。
2.如权利要求1所述的杂合逆转录酶，包含
小鼠白血病病毒逆转录酶(MLVRT)的部分，其包含指和掌结构域，其连接至，
猫白血病病毒逆转录酶(FLVRT)的部分，其包含拇指、连接和RNA酶H结构域。
3.如权利要求2所述的杂合逆转录酶，其中MLVRT的部分与SEQ ID NO：1至少95％相同，并且FLVRT的部分与SEQ ID NO：5至少95％相同。
4.如权利要求2所述的杂合逆转录酶，其中MLVRT的部分包含SEQ ID NO：1，并且FLVRT的部分包含SEQ ID NO：5。
5.如权利要求4所述的杂合逆转录酶，其中FLVRT的部分包含SEQ ID NO：6或SEQ IDNO：10。
6.如权利要求5所述的杂合逆转录酶，其中FLVRT的部分包含SEQ ID NO：7，SEQ ID NO：8，SEQ ID NO：9或SEQ ID NO：11。
7.如权利要求4-6中任一项所述的杂合逆转录酶，其中MLVRT的部分包含SEQ ID NO：2。
8.如权利要求4-6中任一项所述的杂合逆转录酶，其中MLVRT的部分包含SEQ ID NO：3或SEQ ID NO：4。
9.如权利要求4所述的杂合逆转录酶，其中：
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：7；或
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：8；或
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：9；或
MLVRT的部分包含SEQ ID NO：3，并且FLVRT的部分包含SEQ ID NO：11；或
MLVRT的部分包含SEQ ID NO：3，并且FLVRT的部分包含SEQ ID NO：10；或
MLVRT的部分包含SEQ ID NO：4，并且FLVRT的部分包含SEQ ID NO：13。
10.如权利要求4所述的杂合逆转录酶，其中所述杂合逆转录酶包含与SEQ ID NO：14，15，16，17，18，19，34，或35至少95％相同的序列。
11.如权利要求1所述的杂合逆转录酶，包含
猫白血病病毒逆转录酶(FLVRT)的部分，其包含指和掌结构域，其连接至，
小鼠白血病病毒逆转录酶(FLVRT)的部分，其包含RNA酶H结构域。
12.如权利要求11所述的杂合逆转录酶，其中MLVRT的部分包含拇指，连接和RNA酶H结构域。
13.如权利要求12所述的杂合逆转录酶，其中：
FLVRT的部分包含与SEQ ID NO：26至少95％相同的序列；和
MLVRT的部分包含与SEQ ID NO：28至少95％相同的序列。
14.如权利要求12所述的杂合逆转录酶，其中所述杂合逆转录酶包含与SEQ ID NO：30至少95％相同的序列。
15.如权利要求11所述的杂合逆转录酶，其中FLVRT的部分包含指，掌，拇指和连接结构域。
16.如权利要求15所述的杂合逆转录酶，其中：
FLVRT的部分包含与SEQ ID NO：27至少95％相同的序列；和
MLVRT的部分包含与SEQ ID NO：29至少95％相同的序列。
17.如权利要求16所述的杂合逆转录酶，其中所述杂合逆转录酶包含与SEQ ID NO：31至少95％相同的序列。
18.如权利要求1-17中任一项所述的杂合逆转录酶，具有改善热稳定性的至少一种突变。
19.一种包含编码如权利要求1-18中任一项所述的杂合逆转录酶的多核苷酸的核酸。
20.一种反应混合物，包含：
RNA或DNA模板；和
如权利要求1-18中任一项所述的杂合逆转录酶。
21.一种进行逆转录的方法，所述方法包括
在反应混合物中将如权利要求1-18中任一项所述的杂合逆转录酶于一定条件下与模板RNA和引物接触，所述引物与模板RNA杂交，所述条件使所述杂合逆转录酶以模板RNA依赖性方式延伸所述引物以形成cDNA。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112795546A (zh) * 2019-11-13 2021-05-14 中山大学达安基因股份有限公司 一种具有高逆转录效率的耐高温逆转录酶突变体及其应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10961518B2 (en) 2018-01-19 2021-03-30 Bio-Rad Laboratories, Inc. Mutant DNA polymerases

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101613680A (zh) * 2009-01-23 2009-12-30 广州华银医药科技有限公司 莫洛尼氏鼠白血病病毒逆转录酶突变体及其表达方法和应用
CN101684474A (zh) * 2009-02-19 2010-03-31 孙启明 一种重组鼠白血病病毒逆转录酶(mmlv-rt)在大肠杆菌中的可溶性表达及纯化方法
US20140286907A1 (en) * 2011-10-26 2014-09-25 Newvectys Transient expression vectors, preparation and uses thereof
US20160194678A1 (en) * 2005-01-06 2016-07-07 Applied Biosystems, Llc Polypeptides having nucleic acid binding activity and compositions and methods for nucleic acid amplification

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4545985A (en) 1984-01-26 1985-10-08 The United States Of America As Represented By The Secretary, Dept. Of Health And Human Services Pseudomonas exotoxin conjugate immunotoxins
US4751180A (en) 1985-03-28 1988-06-14 Chiron Corporation Expression using fused genes providing for protein product
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US5679543A (en) 1985-08-29 1997-10-21 Genencor International, Inc. DNA sequences, vectors and fusion polypeptides to increase secretion of desired polypeptides from filamentous fungi
US4935233A (en) 1985-12-02 1990-06-19 G. D. Searle And Company Covalently linked polypeptide cell modulators
US5244797B1 (en) 1988-01-13 1998-08-25 Life Technologies Inc Cloned genes encoding reverse transcriptase lacking rnase h activity
DE68924478T2 (de) 1988-12-13 1996-03-07 Univ Colorado Res Prototypische FeLV-Isolate für die Verwendung als Krankheitsmuster oder Impfstoffe.
JPH11271592A (ja) 1998-03-24 1999-10-08 Fuji Photo Optical Co Ltd テレビレンズ用操作装置
US6271004B1 (en) * 1999-06-25 2001-08-07 Display Systems Biotech A/S Method for improved reverse transcription at high temperatures
US20040209276A1 (en) 2002-09-13 2004-10-21 Smith Michael D. Thermostable reverse transcriptases and uses thereof
US7595179B2 (en) 2004-04-19 2009-09-29 Applied Biosystems, Llc Recombinant reverse transcriptases
US10364420B2 (en) * 2015-12-07 2019-07-30 Bio-Rad Laboratories, Inc Dimeric reverse transcriptase

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160194678A1 (en) * 2005-01-06 2016-07-07 Applied Biosystems, Llc Polypeptides having nucleic acid binding activity and compositions and methods for nucleic acid amplification
CN101613680A (zh) * 2009-01-23 2009-12-30 广州华银医药科技有限公司 莫洛尼氏鼠白血病病毒逆转录酶突变体及其表达方法和应用
CN101684474A (zh) * 2009-02-19 2010-03-31 孙启明 一种重组鼠白血病病毒逆转录酶(mmlv-rt)在大肠杆菌中的可溶性表达及纯化方法
US20140286907A1 (en) * 2011-10-26 2014-09-25 Newvectys Transient expression vectors, preparation and uses thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
S K SHARMA等: "Engineering of the human-immunodeficiency-virus-type-1 (HIV-1) reverse transcriptase gene to prevent dimerization of the expressed chimaeric protein: purification and characterization of a monomeric HIV-1 reverse transcriptase", 《BIOTECHNOL APPL BIOCHEM》 *
STEWART等: "Complete genome sequence of two feline leukemia virus subgroup B isolates with novel recomnbination sites", 《GENOME ANNOUNCEMENTS》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112795546A (zh) * 2019-11-13 2021-05-14 中山大学达安基因股份有限公司 一种具有高逆转录效率的耐高温逆转录酶突变体及其应用
CN112795546B (zh) * 2019-11-13 2024-02-27 广州达安基因股份有限公司 一种具有高逆转录效率的耐高温逆转录酶突变体及其应用

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