CN113583122A - 抗人sema4d抗体及其制备方法和应用 - Google Patents
抗人sema4d抗体及其制备方法和应用 Download PDFInfo
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- CN113583122A CN113583122A CN202110860942.7A CN202110860942A CN113583122A CN 113583122 A CN113583122 A CN 113583122A CN 202110860942 A CN202110860942 A CN 202110860942A CN 113583122 A CN113583122 A CN 113583122A
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Abstract
本申请涉及生物技术领域,尤其涉及抗人SEMA4D抗体及其制备方法和应用。所述抗人SEMA4D抗体能特异性地识别SEMA4D蛋白;所述抗人SEMA4D抗体包括第一抗体或第二抗体;所述第一抗体包括SEQ ID NO:1‑SEQ ID NO:37所示氨基酸序列中的任意一种,所述第二抗体序列满足如下至少一条:重链氨基酸序列包括SEQ ID NO:38‑SEQ ID NO:42所示氨基酸序列中的任意一种,轻链氨基酸序列包括SEQ ID NO:43‑SEQ ID NO:47所示氨基酸序列中的任意一种,得到多株亲和力较高的抗人SEMA4D抗体,提供了早期检测诊断标志物及治疗靶点,为联合抗体用药和控制肿瘤提供基础。
Description
技术领域
本申请涉及生物技术领域,尤其涉及抗人SEMA4D抗体及其制备方法和应用。
背景技术
轴突导向蛋白4D(SEMA4D)又称CD100,是轴突导向蛋白N亚族(SEMAs IV)的重要成员之一,其最初作为影响神经发育的轴突导向分子被发现。SEMA4D为I型跨膜蛋白,以同源二聚体形式表达于胞膜,胞膜外区可能存在着不同的功能表位。经过蛋白酶水解作用,SEMA4D能够以分泌型蛋白形式从细胞膜上释放到局部微环境中,可溶性SEMA4D可作为某些疾病的早期诊断标志物。近来越来越多的研究表明,SEMA4D参与了免疫和非免疫系统的多种重要的生理功能,在血管发生及肿瘤生长转移等方面也有重要的生物学功能,并有望成为疾病早期检测诊断标志物及治疗的靶点。
目前公认的SEMA4D的3个受体,分别为Plexin B1、Plexin B2和CD72。Plexin B1与SEMA4D的亲和力最高。研究表明,膜结合型或可溶型的SEMA4D可以和受体Plexin B1结合,激活下游信号通路。SEMA4D分布广泛,在胚胎及成人组织中均有较高表达,其中包括非淋巴组织(心、脑、肾)和淋巴组织(脾、胸腺和淋巴结)。研究表明多种实体肿瘤细胞上SEMA4D也有较高的表达,包括头颈鳞状细胞癌、乳腺癌、前列腺癌、结肠癌、软组织肉瘤和卵巢癌。许多肿瘤细胞上膜型SEMA4D脱落后,可溶型SEMA4D可能以旁分泌形式结合于肿瘤微环境中的内皮细胞上plexin B1,促进新生血管形成,调控肿瘤细胞侵袭转移。
SEMA4D作为靶点的研究与应用显得十分重要,现有SEMA4D的新型人源化单克隆抗体Pepinemab(VX15/2503)是通过杂交瘤技术得到,其分子量较大,单独作用肿瘤细胞的效果不是很明显,联合抗体用药也仅能控制部分患者,因此迫切的需要开发更多优质的SEMA4D抗体来进行研究。
发明内容
本申请提供了抗人SEMA4D抗体及其制备方法和应用,以获得更多种类的SEMA4D抗体,提供更多早期检测诊断标志物及治疗的靶点,为联合抗体用药和控制肿瘤细胞提供研究基础。
第一方面,本申请提供了一种抗人SEMA4D抗体,所述抗人SEMA4D抗体能特异性地识别SEMA4D蛋白;
所述抗人SEMA4D抗体包括第一抗体或第二抗体;
所述第一抗体包括SEQ ID NO:1-SEQ ID NO:37所示氨基酸序列中的任意一种,所述第二抗体序列满足如下至少一条:重链氨基酸序列包括SEQ ID NO:38-SEQ ID NO:42所示氨基酸序列中的任意一种,轻链氨基酸序列包括SEQ ID NO:43-SEQ ID NO:47所示氨基酸序列中的任意一种。
可选的,所述抗人SEMA4D抗体还包括:与所述第一抗体具有同源的氨基酸序列的第一同源抗体,或与所述第二抗体具有同源氨基酸序列的第二同源抗体,其中,
所述第一同源抗体的CDR1区氨基酸序列与所述第一抗体中相对应的CDR1区具有80%以上同源性;
所述第一同源抗体的CDR2区氨基酸序列与所述第一抗体中相对应的CDR2区具有70%以上同源性;
所述第一同源抗体的CDR3区氨基酸序列与所述第一抗体中相对应的CDR3区具有75%以上同源性。
所述第二同源抗体的CDR1区氨基酸序列与所述第二抗体中相对应的CDR1区具有80%以上同源性;
所述第二同源抗体的CDR2区氨基酸序列与所述第二抗体中相对应的CDR2区具有70%以上同源性;
所述所述第二同源抗体的CDR3区氨基酸序列与所述第二抗体中相对应的CDR3区具有75%以上同源性。
可选的,所述第一抗体包括CDR1、CDR2和CDR3中的一种或多种,所述第二抗体包括重链和/或轻链中CDR1、CDR2和CDR3中的一种或多种,其中所述CDR1、所述CDR2和所述CDR3如下表所示:
第二方面,本申请提供了一种制备的抗人SEMA4D抗体的方法,所述方法包括下列步骤:
取正常血清,或用SEMA4D重组蛋白进行多次免疫后得到的血清,分离得到淋巴细胞;
从所述淋巴细胞中提取RNA,进行cDNA的合成,得到cDNA片段;
以所述cDNA为模板,进行PCR扩增,得到基因片段;
将所述基因片段插入载体的DNA序列中,并去除未插入成功的载体,得到含所述基因的载体;
将含所述基因的所述载体感染感受态细胞,进行培养,检测抗人SEMA4D抗体的表达的程度。
一种编码抗人SEMA4D抗体的基因序列,所述基因序列包括第一基因序列和第二基因序列;所述第一基因序列如SEQ ID NO:48-SEQ ID NO:77所示核苷酸序列中的任意一种;所述第二基因序列包括以下至少一条:重链基因如SEQ ID NO:78-SEQ ID NO:82所示的核苷酸序列中任意一种,轻链基因如SEQ ID NO:83-SEQ ID NO:87所示的核苷酸序列中任意一种。
一种含有所述基因序列的载体。
一种转化融合细胞,所述转化融合细胞含有基因序列,或含有载体。
可选的,所述转化融合细胞包括哺乳细胞、酵母细胞、昆虫细胞和原核细胞中任意一种。
一种抗人SEMA4D抗体在诊断标志物及治疗靶点中的应用。
一种SEMA4D蛋白的检测方法,所述方法包括使用所述的抗人SEMA4D抗体。
本申请实施例提供的上述技术方案与现有技术相比具有如下优点:
本申请实施例提供的抗人SEMA4D抗体包括第一抗体和第二抗体,能特异性的识别并结合SEMA4D蛋白,所述抗人SEMA4D抗体包括CDR1区、CDR2区和CDR3区中一种或多种;所述第二抗体包括重链和/或轻链包括CDR1区、CDR2区和CDR3区中一种或多种,得到的多株序列不同且亲和力较高的抗人SEMA4D抗体,提供了更多可以参考的早期检测诊断标志物及治疗的靶点,为联合抗体用药和控制肿瘤细胞提供研究基础。
附图说明
此处的附图被并入说明书中并构成本说明书的一部分,示出了符合本发明的实施例,并与说明书一起用于解释本发明的原理。
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,对于本领域普通技术人员而言,在不付出创造性劳动性的前提下,还可以根据这些附图获得其他的附图。
图1为本申请实施例提供的一种抗人SEMA4D抗体的制备方法的流程示意图;
图2-43为本申请实施例提供的噬菌体上清液进行Flow Cytometry验证结果曲线图;
图44为本申请实施例提供的CSB003、CSB012、CSB044、CSB051重组抗体电泳图;
图45为本申请实施例提供的CSB003、CSB012、CSB044、CSB051重组抗体ELISA验证曲线图;
图46本申请实施例提供的CSB003、CSB012、CSB044、CSB051重组抗体FlowCytometry的曲线结果;
图47本申请提供的SEMA4D阳性抗体CSB-RA835707A1HU与稳转细胞的检测图。
具体实施方式
为使本申请实施例的目的、技术方案和优点更加清楚,下面将结合本申请实施例中的附图,对本申请实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例是本申请的一部分实施例,而不是全部的实施例。基于本申请中的实施例,本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例,都属于本申请保护的范围。
本申请实施例,提供了一种抗人SEMA4D抗体,所述抗人SEMA4D抗体能特异性地识别SEMA4D蛋白;
本申请实施例中,所述SEMA4D蛋白包括CSB-MP835707HU重组蛋白或CSB-MP835707HUd9重组蛋白。
所述抗人SEMA4D抗体包括第一抗体或第二抗体;
所述第一抗体包括SEQ ID NO:1-SEQ ID NO:37所示氨基酸序列中的任意一种,所述第二抗体序列满足如下至少一条:重链氨基酸序列包括SEQ ID NO:38-SEQ ID NO:42所示氨基酸序列中的任意一种,轻链氨基酸序列包括SEQ ID NO:43-SEQ ID NO:47所示氨基酸序列中的任意一种。
本申请实施例中,第一抗体可以来源于羊驼,第二抗体可以来源于鼠。
作为一种可选的实施方式,所述抗人SEMA4D抗体还包括:与所述第一抗体具有同源的氨基酸序列的第一同源抗体,或与所述第二抗体具有同源氨基酸序列的第二同源抗体,其中,
所述第一同源抗体的CDR1区氨基酸序列与所述第一抗体中相对应的CDR1区具有80%以上同源性;
所述第一同源抗体的CDR2区氨基酸序列与所述第一抗体中相对应的CDR2区具有70%以上同源性;
所述第一同源抗体的CDR3区氨基酸序列与所述第一抗体中相对应的CDR3区具有75%以上同源性。
所述第二同源抗体的CDR1区氨基酸序列与所述第二抗体中相对应的CDR1区具有80%以上同源性;
所述第二同源抗体的CDR2区氨基酸序列与所述第二抗体中相对应的CDR2区具有70%以上同源性;
所述所述第二同源抗体的CDR3区氨基酸序列与所述第二抗体中相对应的CDR3区具有75%以上同源性。
本申请实施例中,抗人SEMA4D抗体可以对SEQ ID NO:1-SEQ ID NO:47中任意氨基酸序列的CDR1区、CDR2区和CDR3区进行突变或自然获得,其他部分的序列不变,可以得到多种抗人SEMA4D抗体。
本申请实施例中,第一抗体可以来源于羊驼,第二抗体可以来源于鼠。
本申请实施例中,所述第二抗体包括重链和轻链,重链包括SEQ ID NO:38-SEQ IDNO:42所示氨基酸序列中的任意一种,所述轻链包括SEQ ID NO:43-SEQ ID NO:47所示氨基酸序列中的任意一种,且每条链均包括CDR1区、CDR2区和CDR3区,可以对SEQ ID NO:38-SEQID NO:47中任意氨基酸序列中任意重链和轻链的CDR1区、CDR2区和CDR3区进行突变或自然获得。
作为一种可选的实施方式,所述第一抗体包括CDR1、CDR2和CDR3中的一种或多种,所述第二抗体包括重链和/或轻链中CDR1、CDR2和CDR3中的一种或多种,其中所述CDR1、所述CDR2和所述CDR3如下表所示:
本发明保护的多种抗人SEMA4D抗体包括:以42种抗人SEMA4D抗体为基础,其对应的CDR1区、CDR2区和CDR3区序列如表1、表2和表3所示。
本申请实施例中,SEQ ID NO:1-SEQ ID NO:37所示氨基酸序列的CDR1区、CDR2区和CDR3区氨基酸序列如表1和表2所示。
表1,羊驼天然库FlowCytometry阳性株CDR区序列。
表2,羊驼免疫库FlowCytometry阳性株CDR区序列。
克隆号 | CDR1 | CDR2 | CDR3 |
CSB202 | PYTMG | ATIRFNGNTYYEDSVKG | DNLPWFTEWATLKYAY |
CSB208 | YYAIA | RISSSDGSTVYADSVAG | GLDGDGVEYDY |
CSB209 | RNTMS | TDTNGGSTVYGDSVKG | EALGSYYDLRYLEV |
CSB210 | SYGMS | VINTGDGSTDYADPVKG | PGPGTLVAGPRRLYEFDY |
CSB213 | SYNMG | CITMHDESTYYADSVEG | GCRSYFITPGDDY |
CSB220 | YTAIG | CIVSSDGSTYYADSVKG | SGCEDYGYD |
CSB222 | YYAIG | CISSSDGFTYDADSVKG | GALDWGIRAMCVMAKYDY |
CSB236 | SYAVG | AINWSSISPYAYYADSMKG | NSDYNDVWDARDY |
CSB238 | RYAVG | AISWSGGSPYYSDSMKG | NSDYNDVWDDRDY |
CSB247 | YYNMG | CISSNDGSTYYADSVAG | GCGTGGYDY |
CSB258 | SYAMG | CIVSSDGSTYYADSVEG | SCGEDGYNY |
CSB266 | MPAMG | SFRGGSTNYADFVQG | KALGSYYDLTYLEV |
CSB273 | TYAIG | AISWSGGSPYYADPMKG | NSDYNDVWDERDY |
CSB279 | INAMG | NISSGGSPNYADSVKG | TYGPSSWGMDY |
CSB286 | IKTMG | THTNGGSTNYGDSVKG | EALGSYYDLKYLEV |
CSB294 | SYAMG | AISWSGGSPYYADSMKG | NSDYNDVWDGRDY |
CSB312 | IYPVS | TMTASGNTYYANWANG | GGSTGATYSARAFDP |
CSB315 | VNTIG | CLVSSDGSTYYADSVKG | SCGEDGYDY |
CSB327 | YYAVG | CITNSDGSTYYADSVKG | TGEDGMDCY |
CSB349 | YYNMG | CISSGDGSTYYADSVEG | GCSPFGIPDDY |
CSB356 | SYWMY | GINTDSGSTYYADSVKG | PGAGTGADMVVAGPIGLIQLGS |
CSB364 | YYNIG | CISSGDGSTYYADSVKG | TCGEDGYDY |
CSB372 | DYAIG | CISDSDGSTYYADSVKG | GTVAAEWAYGMDY |
CSB373 | RYTIG | CLSSSDGSTYYADSVKG | GCSPYGINY |
CSB417 | MYVMG | RISLINGSTNYADSVKG | DFRPFGFEY |
CSB420 | SYTMG | ADIRYNGNTYYEDSVKG | DARPWFTEWATVKYTY |
CSB488 | MYVIS | CISSSDGSTYYADSVKG | TCGTDGYDY |
CSB492 | SYWMY | GINSDSGSTYYADSVKG | PGAGAGADMVVAGPIGLIQLGS |
本申请实施例中,所述抗人SEMA4D抗体中所述第二抗体序列包括:重链氨基酸序列包括SEQ ID NO:38-SEQ ID NO:47所示氨基酸序列中的CDR1区、CDR2区和CDR3区氨基酸序列如表3所示。
表3:小鼠免疫库FlowCytometry阳性株CDR区序列。
作为一种可选的实施方式,所述SEMA4D蛋白包括CSB-MP835707HU重组蛋白或CSB-MP835707HUd9重组蛋白。
本申请实施例中,选用CSB-MP835707HU重组蛋白或CSB-MP835707HUd9重组蛋白的都是哺乳表达系统制备,其空间构象更接近天然蛋白。
第二方面,本申请提供了一种制备的抗人SEMA4D抗体的方法,所述方法包括下列步骤:
取天然羊驼的血清,分离淋巴细胞或取SEMA4D重组蛋白,对羊驼或鼠进行多次免疫,分离免疫后羊驼血清或鼠的脾脏,分离淋巴细胞;
从所述淋巴细胞中提取RNA,进行cDNA的合成,得到cDNA片段;
本申请实施例中,取正常羊驼血清,或用SEMA4D重组蛋白进行多次免疫后得到的羊驼血清或小鼠脾脏,分离得到淋巴细胞;
以所述cDNA为模板,进行PCR扩增,得到基因片段,即抗体可变区基因;
将所述基因片段分别与常规pCANTAB5E噬菌体载体连接后,电转化到感受态细胞,分别获得对应的噬菌体库;
使用SEMA4D重组蛋白(CSB-MP835707HU)及293T-SEMA4D稳转细胞筛选出与SEMA4D特异性结合的可变区抗体片段。
本申请实施例中,所述培养包括进行涂板后培养,挑取菌株进行液态培养。
本申请实施例中,抗人SEMA4D抗体使用不同噬菌体展示技术及哺乳系统表达的蛋白筛选出来,由羊驼的纳米抗体基因或小鼠的抗体基因与人IgG4Fc标签融合表达,能够利用所述重组菌株进行稳定表达。其中人IgG4Fc蛋白也可以是其他蛋白标签,或者不带标签,满足不同用途所需。
一种编码抗人SEMA4D抗体的基因序列,所述基因序列包括第一基因序列和第二基因序列;所述第一基因序列如SEQ ID NO:48-SEQ ID NO:84所示核苷酸序列中的任意一种;所述第二基因序列包括以下任意一种:重链基因如SEQ ID NO:85-SEQ ID NO:89所示的核苷酸序列中任意一项,轻链基因如SEQ ID NO:90-SEQ ID NO:94所示的核苷酸序列中任意一项。
本申请实施例中,所述基因序列来源于羊驼和鼠;抗人SEMA4D抗体通过三种不同的噬菌体展示库筛选,并通过哺乳系统表达的SEMA4D蛋白及293T-SEMA4D稳转细胞验证,筛出多株不同序列且亲和力较高的抗体序列,可用作为SEMA4D相关疾病的诊断、鉴定及以SEMA4D为靶点药物研究的原材料。
本申请实施例中,三种不同的噬菌体展示库筛选,包括羊驼天然库基因的噬菌体展示库、羊驼免疫库基因的噬菌体展示库和小鼠免疫库基因的噬菌体展示库。
一种含有所述抗体序列的表达载体。
作为一种可选的实施方式,所述表达载体包括哺乳系统、酵母系统、昆虫系统、原核系统表达。
本申请实施例中,所述表达载为哺乳系统pSecTag2A表达载体。
含有抗体序列,或含有载体的转化融合细胞,所述转化融合细胞包括哺乳细胞、酵母细胞、昆虫细胞和原核细胞中任意一种。
一种抗人SEMA4D抗体在诊断标志物及治疗靶点中的应用。
一种SEMA4D蛋白的检测方法,所述方法包括使用所述的抗人SEMA4D抗体。
本申请实施例中的主要主要实验仪器包括:水平电泳仪(DYY-6C,北京六一)、紫外仪(WD-9403F,北京六一)、台式高速离心机(H1650-W,湘仪)恒温水浴锅(DK-S22,上海精宏)、恒温培养箱(GNP-P270,上海精宏)、洁净工作台(SW-CJ-1FD,苏州安泰)、PCR扩增仪(DL9700,北京东林昌盛)、恒温摇床(TS-100C,上海天呈)、-80℃冰箱(DW-86L628,青岛海尔特种电器有限公司)主要实验试剂包括:Recombinant Human Semaphorin-4D(SEMA4D)(Cusabio,CSB-MP835707HU)、Recombinant Human Semaphorin-4D(SEMA4D)(Cusabio,CSB-MP835707HUd9)、Peroxidase-Affinipure Goat Anti-Human IgG,FcγFragment Specific(Cusabio,CSB-PA160506)、Horseradish Peroxidase-conjugated Streptavidin(Cusabio,CSB-PA813185)、SEMA4D阳性抗体(Cusabio,CSB-RA835707A1HU)、293T-SEMA4D稳转细胞(Cusabio,CSB-SC0005HU)。
实施例1羊驼天然库构建
取新鲜正常羊驼血液样本分离淋巴细胞,提取RNA,用羊驼抗体库引物,从cD NA中扩增出抗体基因,构建羊驼天然库。
实施例2小鼠免疫库构建
取SEMA4D重组蛋白(CSB-MP835707HU)免疫Balb/c小鼠,四次免疫剂量分别为50μg、50μg、50μg、50μg,免疫完成后100μg蛋白冲击,取小鼠脾脏,提取RNA,用小鼠抗体库引物,从cDNA中扩增出抗体基因,构建小鼠免疫库。
实施例3羊驼免疫库构建
取SEMA4D重组蛋白(CSB-MP835707HU)免疫羊驼,四次免疫剂量分别为500μg、250μg、250μg、250μg,免疫完成后取羊驼血液样本分离淋巴细胞,提取RNA,用羊驼抗体库引物,从cDNA中扩增出抗体基因,构建羊驼免疫库。
实施例4序列筛选
取SEMA4D重组蛋白(CSB-MP835707HU)包被到活化后的His Bind Resin上。取拯救后的噬菌体展示抗体库(小鼠免疫库、羊驼天然库或羊驼免疫库)溶液,加入未活化的HisBind Resin,轻微旋转反应1h;1000g离心1min,取上清加入40μL已包被SEMA4D蛋白的HisBind Resin,轻微旋转反应2h;1000g离心1min,去掉上清;加入500μL漂洗缓冲液(含0.1%TritonX-100),重悬,轻微振荡漂洗5min,1000g离心1min,去掉上清,此步骤重复5次;加入200μL洗脱缓冲液,离心取上清,感染TG1菌液,将感染的菌液涂布SOBAG平板。
从SOBAG平板上随机挑取单菌落接种到96孔细菌培养板中,每孔加入2YT-AG培养基,37℃培养过夜;吸取25μL菌液到新的细菌培养板中,30℃诱导过夜;离心,取上清,使用SEMA4D重组蛋白(CSB-MP835707HU)进行ELISA检测。
将上述淘选得到的OD450nm值较大的定为阳性的单克隆,并送武汉金开瑞生物工程有限公司测序,进行阳性克隆进行测序分析,得到的氨基酸序列号1-47所示。
用DNAstar和Clustalw1.8分析抗体的可变区序列,抗体CDR区的氨基酸序列如表1、2、3。
分析的不同序列对应的上清使用293T-SEMA4D稳转细胞进行流式(FlowCytometry)检测,293T细胞阴性上清为对照,如图1-42,横坐标为荧光强度,纵坐标是标准化后的像素的计数。峰分别代表的是不同荧光的强度分布,结果显示,293T-SEMA4D-阴性上清、293T-本申请抗体克隆号上清为对照组,293T-SEMA4D-本申请抗体克隆号上清为实验组,流式细胞仪分析荧光强度如图所示,实验组如293T-SEMA4D-CSB012组等,平均荧光明显强于对照组,说明抗人SEMA4D抗体的阳性上清可结合293T-SEMA4D稳转细胞上表达的SEMA4D蛋白。
实施例5重组表达选择
Flow Cytometry检测的阳性株序列加上所需标签,人工合成核酸序列,连接所需表达载体,转入相应宿主,表达纯化。以pSecTag2A载体为例:选择CSB003、CSB012、CSB044、CSB051测序所得序列加上hIgG4恒定区标签,提交全基因合成,用引物扩增合成产物,插入表达载体pSecTag2A载体中,提取质粒。将质粒转染到293F细胞中,96小时后收集上清,proteinG柱纯化,电泳图如图43,从左到右分别为CSB003重组抗体SDS图、CSB012重组抗体SDS图、CSB044重组抗体SDS图和CSB051重组抗体SDS图,Marker从下自上一依次为::14kDa、18kDa、25kDa、35kDa、45kDa、66kDa、116kDa;经检测其中CSB003放大100ml,纯化后体积为0.9mL,浓度为2.6mg/mL,产量为23.4mg/L;CSB012放大50ml,纯化后体积为2mL,浓度为1mg/mL,产量为40mg/L;CSB044放大200ml,纯化后体积为2.5mL,浓度为0.8mg/mL,产量为10mg/L;CSB051放大100ml,纯化后体积为0.9mL,浓度为2.6mg/mL,产量为23.4mg/L。
实施例6ELISA实验
将SEMA4D重组蛋白(CSB-MP835707HU或CSB-MP835707HUd9)包被2μg/ml,重组表达的SEMA4D抗体梯度稀释后孵育,可以为Peroxidase-Affinipure Goat Anti-Human IgG,FcγFragment Specific(Cusabio,CSB-PA160506)或Horseradish Peroxidase-conjugatedStreptavidin(Cusabio,CSB-PA813185)抗体检测,ELISA检测结果,如图44和表4和表5所示;图中,横坐标代表了抗体浓度,纵坐标代表了吸光值(450nm为最大吸收锋);SEMA4D(CSB-MP835707HU)重组蛋白、SEMA4D(CSB-MP835707HUd9)重组蛋白包板2μg/mL,SEMA4D抗体孵育检测,结果显示SEMA4D抗体都能较好的识别SEMA4D重组蛋白。
对应ELISA检测EC50表4、表5所示。EC50代表了抗体与重组蛋白的亲和力。
表4,SEMA4D(CSB-MP835707HU)蛋白检测结果。
蛋白 | 检测抗体 | EC<sub>50</sub>(ng/mL) |
SEMA4D(CSB-MP835707HU)重组蛋白 | CSB003 | 189.6-291.1 |
SEMA4D(CSB-MP835707HU)重组蛋白 | CSB012 | 90.84-140.6 |
SEMA4D(CSB-MP835707HU)重组蛋白 | CSB044 | 69.79-123.2 |
SEMA4D(CSB-MP835707HU)重组蛋白 | CSB051 | 225.3-326.6 |
表5中SEMA4D(CSB-MP835707HUd9)fc标签,二抗,同时被识别,区别检测。
表5,SEMA4D(CSB-MP835707HUd9)fc标签的蛋白检测结果。
实施例7FlowCytometry实验
取对数生长期的293T、293T-SEMA4D细胞,10%正常非免疫山羊血清封闭后,SEMA4D抗体孵育(2μg/test),FITC-Affinipure Goat Anti-Human IgG,FcγFragmentSpecific抗体检测,流式仪分析。如图45和图46所示,横坐标为荧光强度,纵坐标是标准化后的像素的计数,峰分别代表的是不同荧光的强度分布;293T-SEMA4D阳性抗体及293T-本申请抗体为阴性对照,293T-SEMA4D-SEMA4D阳性抗体为阳性对照,293T-SEMA4D-本申请抗体为实验组,流式细胞仪分析荧光强度如图所示:293T-SEMA4D-SEMA4D阳性抗体荧光强度明显强于293T-SEMA4D阳性抗体,说明293T-SEMA4D稳转细胞上表达了SEMA4D蛋白;293T-SEMA4D-本申请抗体荧光强度明显强于293T细胞-本申请抗体,与293T-SEMA4D-SEMA4D阳性抗体荧光强度差异不大,说明本申请抗体识别293T-SEMA4D稳转细胞上表达的SEMA4D蛋白。
需要说明的是,在本文中,诸如“第一”和“第二”等之类的关系术语仅仅用来将一个实体或者操作与另一个实体或操作区分开来,而不一定要求或者暗示这些实体或操作之间存在任何这种实际的关系或者顺序。而且,术语“包括”、“包含”或者其任何其他变体意在涵盖非排他性的包含,从而使得包括一系列要素的过程、方法、物品或者设备不仅包括那些要素,而且还包括没有明确列出的其他要素,或者是还包括为这种过程、方法、物品或者设备所固有的要素。在没有更多限制的情况下,由语句“包括一个……”限定的要素,并不排除在包括所述要素的过程、方法、物品或者设备中还存在另外的相同要素。
以上所述仅是本发明的具体实施方式,使本领域技术人员能够理解或实现本发明。对这些实施例的多种修改对本领域的技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所申请的原理和新颖特点相一致的最宽的范围。
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100 105 110
Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 9
<211> 124
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Asn Ser Gly Ser
20 25 30
Thr Met Ser Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Ala Ile Ser Asn Thr Ser Gly Asp Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Gly Ser Tyr Leu Asp Tyr Pro Ser Asn Tyr Tyr Ser Tyr Asp
100 105 110
Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 10
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Val Thr Leu Thr Pro Tyr
20 25 30
Thr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Ala Thr Ile Arg Phe Asn Gly Asn Thr Tyr Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asp Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asp Asn Leu Pro Trp Phe Thr Glu Trp Ala Thr Leu Lys Tyr
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 11
<211> 120
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Tyr
20 25 30
Ala Ile Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ser Arg Ile Ser Ser Ser Asp Gly Ser Thr Val Tyr Ala Asp Ser Val
50 55 60
Ala Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Gly Leu Asp Gly Asp Gly Val Glu Tyr Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 12
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Met Val Gln Pro Gly Glu
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Arg Asn
20 25 30
Thr Met Ser Trp Tyr Arg Gln Thr Pro Gly Asn Gln Arg Glu Leu Val
35 40 45
Ala Thr Asp Thr Asn Gly Gly Ser Thr Val Tyr Gly Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Phe Lys His Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn
85 90 95
Ala Glu Ala Leu Gly Ser Tyr Tyr Asp Leu Arg Tyr Leu Glu Val Trp
100 105 110
Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 13
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Val Ile Asn Thr Gly Asp Gly Ser Thr Asp Tyr Ala Asp Pro Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Pro Gly Pro Gly Thr Leu Val Ala Gly Pro Arg Arg Leu Tyr
100 105 110
Glu Phe Asp Tyr Leu Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 14
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asn Ser Tyr
20 25 30
Asn Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Gly Val
35 40 45
Ala Cys Ile Thr Met His Asp Glu Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Glu Gly Arg Ser Thr Ile Ser Arg Asp Asn Thr Lys Asn Thr Met Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Gly Cys Arg Ser Tyr Phe Ile Thr Pro Gly Asp Asp Tyr Trp
100 105 110
Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 15
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Glu
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Thr
20 25 30
Ala Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ser Cys Ile Val Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Trp Cys
85 90 95
Ala Thr Ser Gly Cys Glu Asp Tyr Gly Tyr Asp Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 16
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Met Asp Tyr Tyr
20 25 30
Ala Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ala Ile
35 40 45
Ser Cys Ile Ser Ser Ser Asp Gly Phe Thr Tyr Asp Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Gly Ala Leu Asp Trp Gly Ile Arg Ala Met Cys Val Met Ala
100 105 110
Lys Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 17
<211> 124
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser Tyr
20 25 30
Ala Val Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val
35 40 45
Ala Ala Ile Asn Trp Ser Ser Ile Ser Pro Tyr Ala Tyr Tyr Ala Asp
50 55 60
Ser Met Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr
85 90 95
Ser Cys Ala Ala Asn Ser Asp Tyr Asn Asp Val Trp Asp Ala Arg Asp
100 105 110
Tyr Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 18
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ala Ala Arg Thr Phe Ser Arg Tyr
20 25 30
Ala Val Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Asp Leu Val
35 40 45
Ala Ala Ile Ser Trp Ser Gly Gly Ser Pro Tyr Tyr Ser Asp Ser Met
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Ile Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asn Ser Asp Tyr Asn Asp Val Trp Asp Asp Arg Asp Tyr Trp
100 105 110
Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 19
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Ser Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Tyr
20 25 30
Asn Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Gly Val
35 40 45
Ala Cys Ile Ser Ser Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Ala Gly Arg Phe Thr Ile Ser Arg Asp Asn Thr Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Ser Cys
85 90 95
Ala Thr Gly Cys Gly Thr Gly Gly Tyr Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 20
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser Tyr
20 25 30
Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Pro Glu Gly Val
35 40 45
Ser Cys Ile Val Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Trp Cys
85 90 95
Ala Thr Ser Cys Gly Glu Asp Gly Tyr Asn Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 21
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Met Pro
20 25 30
Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val
35 40 45
Ala Ser Phe Arg Gly Gly Ser Thr Asn Tyr Ala Asp Phe Val Gln Gly
50 55 60
Arg Phe Thr Ile Ser Arg Asp Asp Ala Lys Asn Ala Val Tyr Leu Gln
65 70 75 80
Met Asn Ser Leu Lys Pro Gly Asp Thr Ala Val Tyr Tyr Cys His Ala
85 90 95
Lys Ala Leu Gly Ser Tyr Tyr Asp Leu Thr Tyr Leu Glu Val Trp Gly
100 105 110
Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 22
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr
20 25 30
Ala Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val
35 40 45
Ala Ala Ile Ser Trp Ser Gly Gly Ser Pro Tyr Tyr Ala Asp Pro Met
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Ile Asp Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asn Ser Asp Tyr Asn Asp Val Trp Asp Glu Arg Asp Tyr Trp
100 105 110
Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 23
<211> 119
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Thr Thr Phe Ser Ile Asn
20 25 30
Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Val Glu Leu Val
35 40 45
Ala Asn Ile Ser Ser Gly Gly Ser Pro Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys His
85 90 95
Val Thr Tyr Gly Pro Ser Ser Trp Gly Met Asp Tyr Trp Gly Lys Gly
100 105 110
Thr Gln Val Thr Val Ser Ser
115
<210> 24
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Met Val Gln Pro Gly Glu
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Ser Ile Lys
20 25 30
Thr Met Gly Trp Tyr Arg Gln Thr Pro Gly Lys Gln Arg Glu Leu Val
35 40 45
Ala Thr His Thr Asn Gly Gly Ser Thr Asn Tyr Gly Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Phe Lys His Ala Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn
85 90 95
Ala Glu Ala Leu Gly Ser Tyr Tyr Asp Leu Lys Tyr Leu Glu Val Trp
100 105 110
Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 25
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Ala Ser Ser Tyr
20 25 30
Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Leu
35 40 45
Ala Ala Ile Ser Trp Ser Gly Gly Ser Pro Tyr Tyr Ala Asp Ser Met
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asn Ser Asp Tyr Asn Asp Val Trp Asp Gly Arg Asp Tyr Trp
100 105 110
Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 26
<211> 120
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Gln Ser Val Lys Glu Ser Glu Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ile Tyr Pro
20 25 30
Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly
35 40 45
Thr Met Thr Ala Ser Gly Asn Thr Tyr Tyr Ala Asn Trp Ala Asn Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Met Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Arg Ala Arg Gly Gly
85 90 95
Ser Thr Gly Ala Thr Tyr Ser Ala Arg Ala Phe Asp Pro Trp Gly Pro
100 105 110
Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 27
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ala Cys Glu Ala Ser Gly Leu Ser Phe Ser Val Asn
20 25 30
Thr Ile Gly Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ser Cys Leu Val Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Trp Cys
85 90 95
Ala Arg Ser Cys Gly Glu Asp Gly Tyr Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 28
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Phe Thr Leu Asp Tyr Tyr
20 25 30
Ala Val Gly Trp Phe Arg Arg Ala Pro Gly Lys Glu Pro Glu Ala Val
35 40 45
Ser Cys Ile Thr Asn Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Phe Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Pro Thr Gly Glu Asp Gly Met Asp Cys Tyr Arg Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 29
<211> 120
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Tyr
20 25 30
Asn Met Gly Trp Phe Arg Gln Thr Pro Gly Gln Glu Arg Glu Gly Val
35 40 45
Ala Cys Ile Ser Ser Gly Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Val Gly Cys Ser Pro Phe Gly Ile Pro Asp Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 30
<211> 131
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Trp Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Asn Thr Asp Ser Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asn Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Pro Gly Ala Gly Thr Gly Ala Asp Met Val Val Ala Gly Pro
100 105 110
Ile Gly Leu Ile Gln Leu Gly Ser Arg Gly Gln Gly Thr Gln Val Thr
115 120 125
Val Ser Ser
130
<210> 31
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Tyr
20 25 30
Asn Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Pro Glu Ala Val
35 40 45
Ser Cys Ile Ser Ser Gly Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Arg Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Thr Thr Cys Gly Glu Asp Gly Tyr Asp Tyr Gly Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 32
<211> 122
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr
20 25 30
Ala Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Leu Cys Ile Ser Asp Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Thr Ala Gly Thr Val Ala Ala Glu Trp Ala Tyr Gly Met Asp Tyr Trp
100 105 110
Gly Lys Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 33
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Arg Tyr
20 25 30
Thr Ile Gly Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Gly Val
35 40 45
Ala Cys Leu Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Pro Gly Cys Ser Pro Tyr Gly Ile Asn Tyr Trp Gly Lys Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 34
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Gly Ser Met Tyr
20 25 30
Val Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Asp Trp Val
35 40 45
Ala Arg Ile Ser Leu Ile Asn Gly Ser Thr Asn Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Gly Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Asn Ala Asp Phe Arg Pro Phe Gly Phe Glu Tyr Trp Gly Gln Gly Ser
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 35
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ser Ser Gly Gly Ala Phe Ser Ser Tyr
20 25 30
Thr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ala Ala Asp Ile Arg Tyr Asn Gly Asn Thr Tyr Tyr Glu Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ala Lys Ser Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Gly Asp Ala Arg Pro Trp Phe Thr Glu Trp Ala Thr Val Lys Tyr
100 105 110
Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 36
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Glu Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Met Tyr
20 25 30
Val Ile Ser Trp Phe Arg Gln Ala Pro Gly Lys Glu Pro Glu Gly Val
35 40 45
Ser Cys Ile Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val His
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Trp Cys
85 90 95
Ala Ala Thr Cys Gly Thr Asp Gly Tyr Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 37
<211> 131
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Asp Val Gln Leu Gln Ala Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Trp Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Asn Ser Asp Ser Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asn Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Pro Gly Ala Gly Ala Gly Ala Asp Met Val Val Ala Gly Pro
100 105 110
Ile Gly Leu Ile Gln Leu Gly Ser Arg Gly Gln Gly Thr Gln Val Thr
115 120 125
Val Ser Ser
130
<210> 38
<211> 663
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Cys Ala Gly Gly Thr Gly Cys Ala Gly Cys Thr Gly Ala Ala Gly Cys
1 5 10 15
Ala Gly Thr Cys Thr Gly Gly Gly Gly Cys Thr Gly Ala Gly Cys Thr
20 25 30
Thr Gly Thr Gly Ala Gly Gly Cys Cys Ala Gly Gly Gly Gly Cys Cys
35 40 45
Thr Cys Ala Gly Thr Cys Ala Ala Gly Thr Thr Gly Thr Cys Cys Thr
50 55 60
Gly Cys Ala Cys Ala Gly Cys Thr Thr Cys Thr Gly Gly Cys Thr Thr
65 70 75 80
Thr Ala Ala Cys Ala Thr Thr Ala Ala Ala Gly Ala Cys Gly Ala Cys
85 90 95
Thr Ala Thr Ala Thr Ala Cys Ala Cys Thr Gly Gly Gly Thr Gly Ala
100 105 110
Ala Gly Cys Ala Gly Ala Gly Gly Cys Cys Thr Gly Ala Ala Cys Ala
115 120 125
Gly Gly Gly Cys Cys Thr Gly Gly Ala Gly Thr Gly Gly Ala Thr Thr
130 135 140
Gly Gly Ala Thr Gly Gly Ala Thr Thr Gly Ala Thr Cys Cys Thr Gly
145 150 155 160
Cys Cys Ala Ala Thr Gly Gly Thr Gly Ala Thr Ala Cys Thr Gly Ala
165 170 175
Thr Thr Ala Thr Gly Cys Cys Thr Cys Gly Ala Ala Gly Thr Thr Cys
180 185 190
Cys Ala Gly Gly Gly Cys Ala Ala Gly Gly Cys Cys Ala Cys Thr Ala
195 200 205
Thr Ala Ala Cys Ala Gly Cys Ala Gly Ala Cys Ala Cys Ala Thr Cys
210 215 220
Cys Thr Cys Cys Ala Ala Cys Ala Cys Ala Gly Cys Cys Thr Ala Cys
225 230 235 240
Cys Thr Gly Cys Ala Gly Cys Thr Cys Ala Gly Cys Ala Gly Cys Cys
245 250 255
Thr Gly Ala Cys Ala Thr Cys Thr Gly Ala Gly Gly Ala Cys Ala Cys
260 265 270
Thr Gly Cys Cys Gly Thr Cys Thr Ala Thr Thr Ala Cys Thr Gly Thr
275 280 285
Ala Cys Thr Thr Cys Cys Thr Cys Cys Thr Ala Thr Gly Gly Thr Ala
290 295 300
Ala Cys Thr Ala Cys Gly Gly Ala Gly Cys Thr Ala Thr Gly Gly Ala
305 310 315 320
Cys Thr Ala Cys Thr Gly Gly Gly Gly Thr Cys Ala Ala Gly Gly Ala
325 330 335
Ala Cys Cys Thr Cys Ala Gly Thr Cys Ala Cys Cys Gly Thr Cys Thr
340 345 350
Cys Cys Thr Cys Ala Gly Cys Cys Ala Ala Ala Ala Cys Gly Ala Cys
355 360 365
Ala Cys Cys Cys Cys Cys Ala Thr Cys Thr Gly Thr Cys Thr Ala Thr
370 375 380
Cys Cys Ala Cys Thr Gly Gly Cys Cys Cys Cys Thr Gly Gly Ala Thr
385 390 395 400
Cys Thr Gly Cys Thr Gly Cys Cys Cys Ala Ala Ala Cys Thr Ala Ala
405 410 415
Cys Thr Cys Cys Ala Thr Gly Gly Thr Gly Ala Cys Cys Cys Thr Gly
420 425 430
Gly Gly Ala Thr Gly Cys Cys Thr Gly Gly Thr Cys Ala Ala Gly Gly
435 440 445
Gly Cys Thr Ala Thr Thr Thr Cys Cys Cys Thr Gly Ala Gly Cys Cys
450 455 460
Ala Gly Thr Gly Ala Cys Ala Gly Thr Gly Ala Cys Cys Thr Gly Gly
465 470 475 480
Ala Ala Cys Thr Cys Thr Gly Gly Ala Thr Cys Cys Cys Thr Gly Thr
485 490 495
Cys Cys Ala Gly Cys Gly Gly Thr Gly Thr Gly Cys Ala Cys Ala Cys
500 505 510
Cys Thr Thr Cys Cys Cys Ala Gly Cys Thr Gly Thr Cys Cys Thr Gly
515 520 525
Cys Ala Gly Thr Cys Thr Gly Ala Cys Cys Thr Cys Thr Ala Cys Ala
530 535 540
Cys Thr Cys Thr Gly Ala Gly Cys Ala Gly Cys Thr Cys Ala Gly Thr
545 550 555 560
Gly Ala Cys Thr Gly Thr Cys Cys Cys Cys Thr Cys Cys Ala Gly Cys
565 570 575
Ala Cys Cys Thr Gly Gly Cys Cys Cys Ala Gly Cys Cys Ala Gly Ala
580 585 590
Cys Cys Gly Thr Cys Ala Cys Cys Thr Gly Cys Ala Ala Cys Gly Thr
595 600 605
Thr Gly Cys Cys Cys Ala Cys Cys Cys Gly Gly Cys Cys Ala Gly Cys
610 615 620
Ala Gly Cys Ala Cys Cys Ala Ala Gly Gly Thr Gly Gly Ala Cys Ala
625 630 635 640
Ala Gly Ala Ala Ala Ala Thr Thr Gly Thr Gly Cys Cys Cys Ala Gly
645 650 655
Gly Gly Ala Thr Thr Gly Thr
660
<210> 39
<211> 657
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Gly Ala Gly Gly Thr Gly Ala Ala Gly Cys Thr Gly Gly Thr Gly Gly
1 5 10 15
Ala Gly Thr Cys Ala Gly Gly Ala Cys Cys Thr Gly Gly Cys Cys Thr
20 25 30
Gly Gly Thr Gly Gly Cys Gly Cys Cys Cys Thr Cys Ala Cys Ala Gly
35 40 45
Ala Gly Cys Cys Thr Gly Thr Cys Cys Ala Thr Cys Ala Cys Thr Thr
50 55 60
Gly Cys Ala Cys Thr Gly Thr Cys Thr Cys Thr Gly Gly Gly Thr Thr
65 70 75 80
Thr Thr Cys Ala Thr Thr Ala Ala Gly Cys Ala Gly Cys Thr Ala Thr
85 90 95
Gly Cys Thr Gly Thr Ala Gly Ala Cys Thr Gly Gly Gly Thr Thr Cys
100 105 110
Gly Cys Cys Ala Gly Cys Cys Thr Cys Cys Ala Gly Gly Ala Ala Ala
115 120 125
Gly Gly Gly Thr Cys Thr Gly Gly Ala Gly Thr Gly Gly Cys Thr Gly
130 135 140
Gly Gly Ala Gly Thr Thr Ala Thr Ala Thr Gly Gly Gly Gly Thr Gly
145 150 155 160
Gly Thr Gly Gly Ala Ala Gly Thr Ala Cys Ala Ala Thr Thr Thr Ala
165 170 175
Thr Ala Ala Thr Thr Cys Ala Gly Cys Thr Cys Thr Cys Ala Thr Gly
180 185 190
Thr Cys Cys Ala Gly Ala Cys Thr Gly Ala Gly Cys Ala Thr Cys Ala
195 200 205
Gly Thr Ala Ala Ala Gly Ala Cys Ala Ala Thr Thr Cys Cys Ala Ala
210 215 220
Gly Thr Cys Cys Cys Ala Ala Gly Thr Thr Thr Thr Cys Thr Thr Ala
225 230 235 240
Ala Gly Ala Ala Thr Gly Ala Ala Cys Ala Gly Thr Cys Thr Gly Cys
245 250 255
Ala Ala Ala Cys Thr Gly Ala Thr Gly Ala Cys Ala Cys Ala Gly Cys
260 265 270
Cys Ala Thr Thr Thr Ala Cys Thr Ala Cys Thr Gly Thr Gly Cys Cys
275 280 285
Ala Ala Ala Cys Gly Gly Gly Gly Thr Thr Ala Cys Thr Ala Cys Gly
290 295 300
Gly Cys Thr Ala Thr Thr Thr Ala Gly Ala Cys Thr Cys Cys Thr Gly
305 310 315 320
Gly Gly Gly Cys Cys Ala Ala Gly Gly Cys Ala Cys Cys Ala Cys Thr
325 330 335
Cys Thr Cys Ala Cys Ala Gly Thr Cys Thr Cys Cys Thr Cys Ala Gly
340 345 350
Cys Cys Ala Ala Ala Ala Cys Gly Ala Cys Ala Cys Cys Cys Cys Cys
355 360 365
Ala Thr Cys Thr Gly Thr Cys Thr Ala Thr Cys Cys Ala Cys Thr Gly
370 375 380
Gly Cys Cys Cys Cys Thr Gly Gly Ala Thr Cys Thr Gly Cys Thr Gly
385 390 395 400
Cys Cys Cys Ala Ala Ala Cys Thr Ala Ala Cys Thr Cys Cys Ala Thr
405 410 415
Gly Gly Thr Gly Ala Cys Cys Cys Thr Gly Gly Gly Ala Thr Gly Cys
420 425 430
Cys Thr Gly Gly Thr Cys Ala Ala Gly Gly Gly Cys Thr Ala Thr Thr
435 440 445
Thr Cys Cys Cys Thr Gly Ala Gly Cys Cys Ala Gly Thr Gly Ala Cys
450 455 460
Ala Gly Thr Gly Ala Cys Cys Thr Gly Gly Ala Ala Cys Thr Cys Thr
465 470 475 480
Gly Gly Ala Thr Cys Cys Cys Thr Gly Thr Cys Cys Ala Gly Cys Gly
485 490 495
Gly Thr Gly Thr Gly Cys Ala Cys Ala Cys Cys Thr Thr Cys Cys Cys
500 505 510
Ala Gly Cys Thr Gly Thr Cys Cys Thr Gly Cys Ala Gly Thr Cys Thr
515 520 525
Gly Ala Cys Cys Thr Cys Thr Ala Cys Ala Cys Thr Cys Thr Gly Ala
530 535 540
Gly Cys Ala Gly Cys Thr Cys Ala Gly Thr Gly Ala Cys Thr Gly Thr
545 550 555 560
Cys Cys Cys Cys Thr Cys Cys Ala Gly Cys Ala Cys Cys Thr Gly Gly
565 570 575
Cys Cys Cys Ala Gly Cys Gly Ala Gly Ala Cys Cys Gly Thr Cys Ala
580 585 590
Cys Cys Thr Gly Cys Ala Ala Cys Gly Thr Thr Gly Cys Cys Cys Ala
595 600 605
Cys Cys Cys Gly Gly Cys Cys Ala Gly Cys Ala Gly Cys Ala Cys Cys
610 615 620
Ala Ala Gly Gly Thr Gly Gly Ala Cys Ala Ala Gly Ala Ala Ala Ala
625 630 635 640
Thr Thr Gly Thr Gly Cys Cys Cys Ala Gly Gly Gly Ala Thr Thr Gly
645 650 655
Thr
<210> 40
<211> 663
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Cys Ala Gly Gly Thr Gly Cys Ala Gly Cys Thr Gly Ala Ala Gly Cys
1 5 10 15
Ala Gly Thr Cys Thr Gly Gly Ala Cys Cys Thr Gly Ala Gly Cys Thr
20 25 30
Gly Gly Thr Gly Ala Ala Gly Cys Cys Thr Gly Gly Gly Gly Cys Thr
35 40 45
Thr Cys Ala Gly Thr Gly Ala Ala Gly Ala Thr Gly Thr Cys Cys Thr
50 55 60
Gly Cys Ala Ala Gly Gly Cys Thr Thr Cys Thr Gly Gly Ala Ala Ala
65 70 75 80
Cys Ala Cys Ala Thr Thr Cys Ala Gly Thr Gly Ala Cys Thr Thr Cys
85 90 95
Ala Ala Cys Ala Thr Ala Cys Ala Cys Thr Gly Gly Gly Thr Gly Ala
100 105 110
Ala Gly Cys Ala Gly Ala Ala Cys Cys Ala Thr Gly Gly Ala Ala Ala
115 120 125
Gly Ala Gly Cys Cys Thr Thr Gly Ala Gly Thr Gly Gly Ala Thr Thr
130 135 140
Gly Gly Thr Thr Ala Thr Ala Thr Thr Ala Ala Thr Thr Cys Thr Ala
145 150 155 160
Ala Cys Ala Ala Thr Gly Gly Thr Gly Gly Thr Ala Cys Thr Ala Gly
165 170 175
Cys Thr Ala Cys Ala Ala Cys Cys Ala Gly Ala Ala Gly Thr Thr Cys
180 185 190
Ala Ala Gly Gly Ala Cys Ala Ala Gly Gly Cys Cys Ala Cys Ala Thr
195 200 205
Thr Gly Ala Cys Thr Gly Thr Ala Ala Ala Cys Ala Ala Gly Thr Cys
210 215 220
Cys Thr Cys Cys Ala Gly Cys Ala Cys Ala Gly Cys Cys Thr Ala Cys
225 230 235 240
Ala Thr Gly Gly Ala Gly Cys Thr Cys Cys Gly Cys Ala Gly Cys Cys
245 250 255
Thr Gly Ala Cys Ala Thr Cys Gly Gly Ala Gly Gly Ala Thr Thr Cys
260 265 270
Thr Gly Cys Ala Gly Thr Cys Thr Thr Thr Thr Ala Cys Thr Gly Thr
275 280 285
Gly Gly Gly Ala Cys Gly Gly Gly Thr Gly Ala Thr Thr Ala Cys Gly
290 295 300
Ala Ala Gly Gly Gly Thr Thr Thr Gly Cys Thr Ala Thr Gly Gly Ala
305 310 315 320
Cys Thr Ala Cys Thr Gly Gly Gly Gly Thr Cys Ala Ala Gly Gly Ala
325 330 335
Ala Cys Cys Thr Cys Ala Gly Thr Cys Ala Cys Cys Gly Thr Cys Thr
340 345 350
Cys Cys Thr Cys Ala Gly Cys Cys Ala Ala Ala Ala Cys Gly Ala Cys
355 360 365
Ala Cys Cys Cys Cys Cys Ala Thr Cys Thr Gly Thr Cys Thr Ala Thr
370 375 380
Cys Cys Ala Cys Thr Gly Gly Cys Cys Cys Cys Thr Gly Gly Ala Thr
385 390 395 400
Cys Thr Gly Cys Thr Gly Cys Cys Cys Ala Ala Ala Cys Thr Ala Ala
405 410 415
Cys Thr Cys Cys Ala Thr Gly Gly Thr Gly Ala Cys Cys Cys Thr Gly
420 425 430
Gly Gly Ala Thr Gly Cys Cys Thr Gly Gly Thr Cys Ala Ala Gly Gly
435 440 445
Gly Cys Thr Ala Thr Thr Thr Cys Cys Cys Thr Gly Ala Gly Cys Cys
450 455 460
Ala Gly Thr Gly Ala Cys Ala Gly Thr Gly Ala Cys Cys Thr Gly Gly
465 470 475 480
Ala Ala Cys Thr Cys Thr Gly Gly Ala Thr Cys Cys Cys Thr Gly Thr
485 490 495
Cys Cys Ala Gly Cys Gly Gly Thr Gly Thr Gly Cys Ala Cys Ala Cys
500 505 510
Cys Thr Thr Cys Cys Cys Ala Gly Cys Thr Gly Thr Cys Cys Thr Gly
515 520 525
Cys Ala Gly Thr Cys Thr Gly Ala Cys Cys Thr Cys Thr Ala Cys Ala
530 535 540
Cys Thr Cys Thr Gly Ala Gly Cys Ala Gly Cys Thr Cys Ala Gly Thr
545 550 555 560
Gly Ala Cys Thr Gly Thr Cys Cys Cys Cys Thr Cys Cys Ala Gly Cys
565 570 575
Ala Cys Cys Thr Gly Gly Cys Cys Cys Ala Gly Cys Cys Ala Gly Ala
580 585 590
Cys Cys Gly Thr Cys Ala Cys Cys Thr Gly Cys Ala Ala Cys Gly Thr
595 600 605
Thr Gly Cys Cys Cys Ala Cys Cys Cys Gly Gly Cys Cys Ala Gly Cys
610 615 620
Ala Gly Cys Ala Cys Cys Ala Ala Ala Gly Thr Gly Gly Ala Cys Ala
625 630 635 640
Ala Gly Ala Ala Ala Ala Thr Thr Gly Thr Gly Cys Cys Cys Ala Gly
645 650 655
Gly Gly Ala Thr Thr Gly Thr
660
<210> 41
<211> 657
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Cys Ala Gly Gly Thr Gly Cys Ala Gly Cys Thr Gly Ala Ala Gly Cys
1 5 10 15
Ala Gly Thr Cys Ala Gly Gly Ala Cys Cys Thr Gly Gly Cys Cys Thr
20 25 30
Gly Gly Thr Gly Gly Cys Gly Cys Cys Cys Thr Cys Ala Cys Ala Gly
35 40 45
Ala Gly Cys Cys Thr Gly Thr Cys Cys Ala Thr Cys Ala Cys Thr Thr
50 55 60
Gly Cys Ala Cys Thr Gly Thr Cys Thr Cys Thr Gly Gly Gly Thr Thr
65 70 75 80
Thr Thr Cys Ala Thr Thr Ala Ala Cys Cys Ala Gly Cys Thr Ala Thr
85 90 95
Gly Cys Thr Gly Thr Ala Gly Ala Cys Thr Gly Gly Gly Thr Thr Cys
100 105 110
Gly Cys Cys Ala Gly Cys Cys Thr Cys Cys Ala Gly Gly Ala Ala Ala
115 120 125
Gly Gly Ala Thr Cys Thr Gly Gly Ala Gly Thr Gly Gly Cys Thr Gly
130 135 140
Gly Gly Ala Gly Thr Ala Ala Thr Ala Thr Gly Gly Gly Gly Thr Gly
145 150 155 160
Gly Thr Gly Gly Ala Ala Gly Thr Ala Cys Ala Ala Thr Thr Thr Ala
165 170 175
Thr Ala Ala Thr Thr Cys Ala Gly Cys Thr Cys Thr Cys Ala Thr Gly
180 185 190
Thr Cys Cys Ala Gly Ala Cys Thr Gly Ala Gly Cys Ala Thr Cys Ala
195 200 205
Gly Thr Ala Ala Ala Gly Ala Cys Ala Ala Cys Thr Cys Cys Ala Ala
210 215 220
Gly Ala Gly Cys Cys Ala Ala Gly Thr Thr Thr Thr Cys Thr Thr Ala
225 230 235 240
Ala Ala Ala Ala Thr Gly Ala Ala Cys Ala Gly Thr Cys Thr Cys Cys
245 250 255
Ala Ala Ala Cys Thr Gly Ala Thr Gly Ala Cys Ala Cys Ala Gly Cys
260 265 270
Cys Ala Thr Thr Thr Ala Cys Thr Ala Cys Thr Gly Thr Gly Cys Thr
275 280 285
Ala Ala Ala Cys Gly Cys Gly Gly Gly Thr Ala Cys Thr Ala Cys Gly
290 295 300
Gly Thr Thr Thr Thr Cys Thr Thr Gly Ala Cys Thr Ala Cys Thr Gly
305 310 315 320
Gly Gly Gly Cys Cys Ala Ala Gly Gly Cys Ala Cys Cys Ala Cys Thr
325 330 335
Cys Thr Cys Ala Cys Ala Gly Thr Cys Thr Cys Cys Thr Cys Ala Gly
340 345 350
Cys Cys Ala Ala Ala Ala Cys Gly Ala Cys Ala Cys Cys Cys Cys Cys
355 360 365
Ala Thr Cys Thr Gly Thr Cys Thr Ala Thr Cys Cys Ala Cys Thr Gly
370 375 380
Gly Cys Cys Cys Cys Thr Gly Gly Ala Thr Cys Thr Gly Cys Thr Gly
385 390 395 400
Cys Cys Cys Ala Ala Ala Cys Thr Ala Ala Cys Thr Cys Cys Ala Thr
405 410 415
Gly Gly Thr Gly Ala Cys Cys Cys Thr Gly Gly Gly Ala Thr Gly Cys
420 425 430
Cys Thr Gly Gly Thr Cys Ala Ala Gly Gly Gly Cys Thr Ala Thr Thr
435 440 445
Thr Cys Cys Cys Thr Gly Ala Gly Cys Cys Ala Gly Thr Gly Ala Cys
450 455 460
Ala Gly Thr Gly Ala Cys Cys Thr Gly Gly Ala Ala Cys Thr Cys Thr
465 470 475 480
Gly Gly Ala Thr Cys Cys Cys Thr Gly Thr Cys Cys Ala Gly Cys Gly
485 490 495
Gly Thr Gly Thr Gly Cys Ala Cys Ala Cys Cys Thr Thr Cys Cys Cys
500 505 510
Ala Gly Cys Thr Gly Thr Cys Cys Thr Gly Cys Ala Gly Thr Cys Thr
515 520 525
Gly Ala Cys Cys Thr Cys Thr Ala Cys Ala Cys Thr Cys Thr Gly Ala
530 535 540
Gly Cys Ala Gly Cys Thr Cys Ala Gly Thr Gly Ala Cys Thr Gly Thr
545 550 555 560
Cys Cys Cys Cys Thr Cys Cys Ala Gly Cys Ala Cys Cys Thr Gly Gly
565 570 575
Cys Cys Cys Ala Gly Cys Gly Ala Gly Ala Cys Cys Gly Thr Cys Ala
580 585 590
Cys Cys Thr Gly Cys Ala Ala Cys Gly Thr Thr Gly Cys Cys Cys Ala
595 600 605
Cys Cys Cys Gly Gly Cys Cys Ala Gly Cys Ala Gly Cys Ala Cys Cys
610 615 620
Ala Ala Gly Gly Thr Gly Gly Ala Cys Ala Ala Gly Ala Ala Ala Ala
625 630 635 640
Thr Thr Gly Thr Gly Cys Cys Cys Ala Gly Gly Gly Ala Thr Thr Gly
645 650 655
Thr
<210> 42
<211> 666
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Cys Ala Gly Gly Thr Gly Cys Ala Gly Cys Thr Gly Ala Ala Gly Cys
1 5 10 15
Ala Gly Thr Cys Thr Gly Gly Ala Gly Cys Thr Gly Ala Gly Cys Thr
20 25 30
Gly Gly Thr Thr Ala Gly Gly Cys Cys Thr Gly Gly Gly Ala Cys Thr
35 40 45
Thr Cys Ala Gly Thr Gly Ala Ala Gly Ala Thr Gly Thr Cys Cys Thr
50 55 60
Gly Cys Ala Ala Gly Gly Cys Thr Gly Cys Thr Gly Gly Ala Thr Ala
65 70 75 80
Cys Ala Gly Cys Thr Thr Cys Ala Cys Thr Thr Ala Cys Thr Ala Cys
85 90 95
Thr Gly Gly Ala Thr Ala Ala Gly Thr Thr Gly Gly Gly Thr Ala Ala
100 105 110
Ala Ala Cys Ala Gly Ala Gly Gly Cys Cys Thr Gly Gly Ala Cys Ala
115 120 125
Thr Gly Gly Cys Cys Thr Thr Gly Ala Gly Thr Gly Gly Ala Thr Thr
130 135 140
Gly Gly Ala Gly Ala Thr Ala Thr Thr Thr Ala Cys Cys Cys Thr Gly
145 150 155 160
Gly Ala Gly Gly Thr Gly Thr Thr Thr Ala Thr Ala Cys Thr Ala Ala
165 170 175
Cys Thr Ala Cys Ala Ala Thr Gly Ala Gly Ala Ala Gly Thr Thr Cys
180 185 190
Ala Ala Gly Gly Gly Cys Ala Ala Gly Gly Cys Cys Ala Cys Ala Cys
195 200 205
Thr Gly Ala Cys Thr Gly Cys Ala Gly Ala Cys Ala Cys Ala Thr Cys
210 215 220
Cys Thr Cys Cys Ala Gly Cys Ala Cys Ala Gly Cys Cys Thr Ala Cys
225 230 235 240
Ala Thr Ala Cys Ala Ala Cys Thr Cys Ala Cys Cys Ala Gly Cys Cys
245 250 255
Thr Gly Ala Cys Ala Thr Cys Thr Gly Ala Gly Gly Ala Cys Thr Cys
260 265 270
Thr Gly Cys Cys Ala Thr Cys Thr Ala Thr Thr Ala Thr Thr Gly Thr
275 280 285
Gly Cys Ala Gly Ala Gly Gly Gly Ala Cys Ala Thr Thr Ala Cys Gly
290 295 300
Ala Ala Ala Gly Gly Gly Gly Gly Gly Gly Gly Ala Cys Thr Thr Thr
305 310 315 320
Thr Gly Ala Cys Cys Ala Cys Thr Gly Gly Gly Gly Cys Cys Ala Ala
325 330 335
Gly Gly Cys Ala Cys Cys Ala Cys Thr Cys Thr Cys Ala Cys Ala Gly
340 345 350
Thr Cys Thr Cys Cys Thr Cys Ala Gly Cys Cys Ala Ala Ala Ala Cys
355 360 365
Gly Ala Cys Ala Cys Cys Cys Cys Cys Ala Thr Cys Thr Gly Thr Cys
370 375 380
Thr Ala Thr Cys Cys Ala Cys Thr Gly Gly Cys Cys Cys Cys Thr Gly
385 390 395 400
Gly Ala Thr Cys Thr Gly Cys Thr Gly Cys Cys Cys Ala Ala Ala Cys
405 410 415
Thr Ala Ala Cys Thr Cys Cys Ala Thr Gly Gly Thr Gly Ala Cys Cys
420 425 430
Cys Thr Gly Gly Gly Ala Thr Gly Cys Cys Thr Gly Gly Thr Cys Ala
435 440 445
Ala Gly Gly Gly Cys Thr Ala Thr Thr Thr Cys Cys Cys Thr Gly Ala
450 455 460
Gly Cys Cys Ala Gly Thr Gly Ala Cys Ala Gly Thr Gly Ala Cys Cys
465 470 475 480
Thr Gly Gly Ala Ala Cys Thr Cys Thr Gly Gly Ala Thr Cys Cys Cys
485 490 495
Thr Gly Thr Cys Cys Ala Gly Cys Gly Gly Thr Gly Thr Gly Cys Ala
500 505 510
Cys Ala Cys Cys Thr Thr Cys Cys Cys Ala Gly Cys Thr Gly Thr Cys
515 520 525
Cys Thr Gly Cys Ala Gly Thr Cys Thr Gly Ala Cys Cys Thr Cys Thr
530 535 540
Ala Cys Ala Cys Thr Cys Thr Gly Ala Gly Cys Ala Gly Cys Thr Cys
545 550 555 560
Ala Gly Thr Gly Ala Cys Thr Gly Thr Cys Cys Cys Cys Thr Cys Cys
565 570 575
Ala Gly Cys Ala Cys Cys Thr Gly Gly Cys Cys Cys Ala Gly Cys Cys
580 585 590
Ala Gly Ala Cys Cys Gly Thr Cys Ala Cys Cys Thr Gly Cys Ala Ala
595 600 605
Cys Gly Thr Thr Gly Cys Cys Cys Ala Cys Cys Cys Gly Gly Cys Cys
610 615 620
Ala Gly Cys Ala Gly Cys Ala Cys Cys Ala Ala Gly Gly Thr Gly Gly
625 630 635 640
Ala Cys Ala Ala Gly Ala Ala Ala Ala Thr Thr Gly Thr Gly Cys Cys
645 650 655
Cys Ala Gly Gly Gly Ala Thr Thr Gly Thr
660 665
<210> 43
<211> 645
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Cys Ala Ala Ala Thr Thr Gly Thr Thr Cys Thr Cys Ala Cys Cys Cys
1 5 10 15
Ala Gly Thr Cys Thr Cys Cys Ala Gly Cys Ala Cys Thr Cys Ala Thr
20 25 30
Gly Gly Cys Thr Gly Cys Ala Thr Cys Thr Cys Cys Ala Gly Gly Gly
35 40 45
Gly Ala Gly Ala Ala Gly Gly Thr Cys Ala Cys Cys Ala Thr Cys Ala
50 55 60
Cys Cys Thr Gly Cys Ala Gly Thr Gly Thr Cys Ala Gly Cys Thr Cys
65 70 75 80
Ala Ala Gly Thr Ala Thr Ala Ala Gly Thr Thr Cys Cys Ala Gly Cys
85 90 95
Ala Ala Cys Thr Thr Ala Cys Ala Cys Thr Gly Gly Thr Ala Cys Cys
100 105 110
Ala Gly Cys Ala Gly Ala Ala Gly Thr Cys Ala Gly Gly Ala Ala Cys
115 120 125
Cys Thr Cys Cys Cys Cys Cys Ala Ala Ala Cys Cys Cys Thr Thr Gly
130 135 140
Ala Thr Thr Thr Ala Thr Gly Gly Cys Ala Cys Ala Thr Cys Cys Ala
145 150 155 160
Ala Cys Cys Thr Thr Gly Cys Thr Thr Cys Thr Gly Gly Ala Gly Thr
165 170 175
Cys Cys Cys Thr Gly Thr Thr Cys Gly Cys Thr Thr Cys Ala Gly Thr
180 185 190
Gly Gly Cys Ala Gly Thr Gly Gly Ala Thr Cys Thr Gly Gly Gly Ala
195 200 205
Cys Cys Thr Cys Thr Thr Ala Thr Thr Cys Thr Cys Thr Cys Ala Cys
210 215 220
Ala Ala Thr Cys Ala Gly Cys Ala Gly Cys Ala Thr Gly Gly Ala Gly
225 230 235 240
Gly Cys Thr Gly Ala Ala Gly Ala Thr Gly Cys Thr Gly Cys Cys Thr
245 250 255
Cys Thr Thr Ala Thr Thr Ala Cys Thr Gly Thr Cys Ala Ala Cys Ala
260 265 270
Gly Thr Gly Gly Ala Gly Thr Ala Gly Thr Thr Ala Cys Cys Cys Gly
275 280 285
Thr Ala Thr Ala Cys Gly Thr Thr Cys Gly Gly Ala Thr Cys Gly Gly
290 295 300
Gly Gly Ala Cys Cys Ala Ala Gly Cys Thr Gly Gly Ala Ala Ala Thr
305 310 315 320
Ala Ala Ala Ala Cys Gly Gly Gly Cys Thr Gly Ala Thr Gly Cys Thr
325 330 335
Gly Cys Ala Cys Cys Ala Ala Cys Thr Gly Thr Ala Thr Cys Cys Ala
340 345 350
Thr Cys Thr Thr Cys Cys Cys Ala Cys Cys Ala Thr Cys Cys Ala Gly
355 360 365
Thr Gly Ala Gly Cys Ala Gly Thr Thr Ala Ala Cys Ala Thr Cys Thr
370 375 380
Gly Gly Ala Gly Gly Thr Gly Cys Cys Thr Cys Ala Gly Thr Cys Gly
385 390 395 400
Thr Gly Thr Gly Cys Thr Thr Cys Thr Thr Gly Ala Ala Cys Ala Ala
405 410 415
Cys Thr Thr Cys Thr Ala Cys Cys Cys Cys Ala Ala Ala Gly Ala Cys
420 425 430
Ala Thr Cys Ala Ala Thr Gly Thr Cys Ala Ala Gly Thr Gly Gly Ala
435 440 445
Ala Gly Ala Thr Thr Gly Ala Thr Gly Gly Cys Ala Gly Thr Gly Ala
450 455 460
Ala Cys Gly Ala Cys Ala Ala Ala Ala Thr Gly Gly Cys Gly Thr Cys
465 470 475 480
Cys Thr Gly Ala Ala Cys Ala Gly Thr Thr Gly Gly Ala Cys Thr Gly
485 490 495
Ala Thr Cys Ala Gly Gly Ala Cys Ala Gly Cys Ala Ala Ala Gly Ala
500 505 510
Cys Ala Gly Cys Ala Cys Cys Thr Ala Cys Ala Gly Cys Ala Thr Gly
515 520 525
Ala Gly Cys Ala Gly Cys Ala Cys Cys Cys Thr Cys Ala Cys Gly Thr
530 535 540
Thr Gly Ala Cys Cys Ala Ala Gly Gly Ala Cys Gly Ala Gly Thr Ala
545 550 555 560
Thr Gly Ala Ala Cys Gly Ala Cys Ala Thr Ala Ala Cys Ala Gly Cys
565 570 575
Thr Ala Thr Ala Cys Cys Thr Gly Thr Gly Ala Gly Gly Cys Cys Ala
580 585 590
Cys Thr Cys Ala Cys Ala Ala Gly Ala Cys Ala Thr Cys Ala Ala Cys
595 600 605
Thr Thr Cys Ala Cys Cys Cys Ala Thr Thr Gly Thr Cys Ala Ala Gly
610 615 620
Ala Gly Cys Thr Thr Cys Ala Ala Cys Ala Gly Gly Ala Ala Thr Gly
625 630 635 640
Ala Gly Thr Gly Thr
645
<210> 44
<211> 639
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Gly Ala Thr Ala Thr Thr Gly Thr Gly Ala Thr Gly Ala Cys Gly Cys
1 5 10 15
Ala Gly Gly Cys Thr Cys Cys Ala Gly Cys Ala Ala Thr Cys Ala Thr
20 25 30
Gly Thr Cys Thr Gly Cys Ala Thr Cys Thr Cys Cys Ala Gly Gly Gly
35 40 45
Gly Ala Gly Ala Ala Gly Gly Thr Cys Ala Cys Cys Ala Thr Gly Ala
50 55 60
Cys Cys Thr Gly Cys Ala Gly Thr Gly Cys Cys Ala Cys Cys Thr Cys
65 70 75 80
Ala Ala Gly Thr Gly Thr Ala Ala Gly Thr Thr Ala Cys Ala Thr Ala
85 90 95
Thr Ala Cys Thr Gly Gly Thr Ala Cys Cys Ala Gly Cys Ala Gly Ala
100 105 110
Ala Gly Cys Cys Ala Gly Gly Ala Thr Cys Cys Thr Cys Cys Cys Cys
115 120 125
Cys Ala Gly Ala Cys Thr Cys Cys Thr Gly Ala Thr Thr Thr Ala Thr
130 135 140
Gly Ala Cys Ala Cys Ala Thr Cys Cys Ala Cys Cys Cys Thr Gly Gly
145 150 155 160
Cys Thr Thr Cys Thr Gly Gly Ala Gly Thr Cys Cys Cys Thr Gly Thr
165 170 175
Thr Cys Gly Cys Thr Thr Cys Ala Gly Thr Gly Gly Cys Ala Gly Thr
180 185 190
Gly Gly Gly Thr Cys Thr Gly Gly Gly Ala Cys Cys Thr Cys Thr Thr
195 200 205
Ala Cys Thr Cys Thr Cys Thr Cys Ala Cys Ala Ala Thr Cys Ala Gly
210 215 220
Cys Cys Gly Ala Ala Thr Gly Gly Ala Gly Gly Cys Thr Gly Ala Ala
225 230 235 240
Gly Ala Thr Gly Cys Thr Gly Cys Cys Ala Cys Thr Thr Ala Thr Thr
245 250 255
Ala Cys Thr Gly Cys Cys Ala Gly Cys Ala Gly Thr Gly Gly Ala Gly
260 265 270
Thr Ala Gly Thr Thr Ala Cys Cys Cys Ala Cys Thr Cys Ala Cys Gly
275 280 285
Thr Thr Cys Gly Gly Thr Gly Cys Thr Gly Gly Gly Ala Cys Cys Ala
290 295 300
Ala Gly Cys Thr Gly Gly Ala Gly Cys Thr Gly Ala Ala Ala Cys Gly
305 310 315 320
Gly Gly Cys Thr Gly Ala Thr Gly Cys Thr Gly Cys Ala Cys Cys Ala
325 330 335
Ala Cys Thr Gly Thr Ala Thr Cys Cys Ala Thr Cys Thr Thr Cys Cys
340 345 350
Cys Ala Cys Cys Ala Thr Cys Cys Ala Gly Thr Gly Ala Gly Cys Ala
355 360 365
Gly Thr Thr Ala Ala Cys Ala Thr Cys Thr Gly Gly Ala Gly Gly Thr
370 375 380
Gly Cys Cys Thr Cys Ala Gly Thr Cys Gly Thr Gly Thr Gly Cys Thr
385 390 395 400
Thr Cys Thr Thr Gly Ala Ala Cys Ala Ala Cys Thr Thr Cys Thr Ala
405 410 415
Cys Cys Cys Cys Ala Ala Ala Gly Ala Cys Ala Thr Cys Ala Ala Thr
420 425 430
Gly Thr Cys Ala Ala Gly Thr Gly Gly Ala Ala Gly Ala Thr Thr Gly
435 440 445
Ala Thr Gly Gly Cys Ala Gly Thr Gly Ala Ala Cys Gly Ala Cys Ala
450 455 460
Ala Ala Ala Thr Gly Gly Cys Gly Thr Cys Cys Thr Gly Ala Ala Cys
465 470 475 480
Ala Gly Thr Thr Gly Gly Ala Cys Thr Gly Ala Thr Cys Ala Gly Gly
485 490 495
Ala Cys Ala Gly Cys Ala Ala Ala Gly Ala Cys Ala Gly Cys Ala Cys
500 505 510
Cys Thr Ala Cys Ala Gly Cys Ala Thr Gly Ala Gly Cys Ala Gly Cys
515 520 525
Ala Cys Cys Cys Thr Cys Ala Cys Gly Thr Thr Gly Ala Cys Cys Ala
530 535 540
Ala Gly Gly Ala Cys Gly Ala Gly Thr Ala Thr Gly Ala Ala Cys Gly
545 550 555 560
Ala Cys Ala Thr Ala Ala Cys Ala Gly Cys Thr Ala Thr Ala Cys Cys
565 570 575
Thr Gly Thr Gly Ala Gly Gly Cys Cys Ala Cys Thr Cys Ala Cys Ala
580 585 590
Ala Gly Ala Cys Ala Thr Cys Ala Ala Cys Thr Thr Cys Ala Cys Cys
595 600 605
Cys Ala Thr Thr Gly Thr Cys Ala Ala Gly Ala Gly Cys Thr Thr Cys
610 615 620
Ala Ala Cys Ala Gly Gly Ala Ala Thr Gly Ala Gly Thr Gly Thr
625 630 635
<210> 45
<211> 642
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Gly Ala Cys Ala Thr Thr Gly Thr Gly Ala Thr Gly Ala Cys Cys Cys
1 5 10 15
Ala Gly Thr Cys Thr Cys Cys Ala Gly Cys Thr Thr Cys Ala Cys Thr
20 25 30
Gly Thr Cys Thr Gly Cys Ala Thr Cys Thr Gly Thr Gly Gly Gly Ala
35 40 45
Gly Ala Ala Ala Cys Thr Gly Thr Cys Ala Cys Cys Ala Thr Cys Ala
50 55 60
Cys Ala Thr Gly Thr Gly Gly Ala Gly Cys Ala Ala Gly Thr Gly Ala
65 70 75 80
Gly Ala Ala Thr Cys Thr Thr Thr Ala Cys Gly Gly Thr Gly Cys Thr
85 90 95
Thr Thr Ala Ala Ala Thr Thr Gly Gly Thr Ala Thr Cys Ala Gly Cys
100 105 110
Thr Gly Ala Ala Ala Cys Ala Gly Gly Gly Ala Ala Ala Ala Thr Cys
115 120 125
Thr Cys Cys Thr Cys Ala Gly Cys Thr Cys Cys Thr Gly Ala Thr Cys
130 135 140
Thr Ala Thr Gly Gly Thr Gly Cys Gly Ala Cys Cys Ala Ala Cys Thr
145 150 155 160
Thr Gly Gly Cys Ala Gly Ala Thr Gly Gly Cys Ala Thr Gly Thr Cys
165 170 175
Ala Thr Cys Gly Ala Gly Gly Thr Thr Cys Ala Gly Ala Gly Gly Cys
180 185 190
Ala Gly Thr Gly Gly Ala Thr Thr Thr Gly Gly Thr Ala Gly Ala Cys
195 200 205
Ala Gly Thr Ala Thr Thr Cys Thr Cys Thr Cys Ala Ala Gly Ala Thr
210 215 220
Cys Ala Gly Thr Ala Ala Cys Cys Thr Gly Cys Ala Thr Cys Cys Thr
225 230 235 240
Gly Ala Cys Gly Ala Thr Gly Thr Thr Gly Cys Ala Ala Thr Gly Thr
245 250 255
Ala Thr Thr Ala Thr Thr Gly Thr Cys Ala Ala Ala Ala Thr Gly Thr
260 265 270
Gly Thr Thr Ala Ala Cys Thr Ala Cys Thr Cys Cys Gly Cys Thr Cys
275 280 285
Ala Cys Gly Thr Thr Cys Gly Gly Thr Gly Cys Thr Gly Gly Gly Ala
290 295 300
Cys Cys Ala Ala Gly Cys Thr Gly Gly Ala Gly Cys Thr Gly Ala Ala
305 310 315 320
Ala Cys Gly Gly Gly Cys Thr Gly Ala Thr Gly Cys Thr Gly Cys Ala
325 330 335
Cys Cys Ala Ala Cys Thr Gly Thr Ala Thr Cys Cys Ala Thr Cys Thr
340 345 350
Thr Cys Cys Cys Ala Cys Cys Ala Thr Cys Cys Ala Gly Thr Gly Ala
355 360 365
Gly Cys Ala Gly Thr Thr Ala Ala Cys Ala Thr Cys Thr Gly Gly Ala
370 375 380
Gly Gly Thr Gly Cys Cys Thr Cys Ala Gly Thr Cys Gly Thr Gly Thr
385 390 395 400
Gly Cys Thr Thr Cys Thr Thr Gly Ala Ala Cys Ala Ala Cys Thr Thr
405 410 415
Cys Thr Ala Cys Cys Cys Cys Ala Ala Ala Gly Ala Cys Ala Thr Cys
420 425 430
Ala Ala Thr Gly Thr Cys Ala Ala Gly Thr Gly Gly Ala Ala Gly Ala
435 440 445
Thr Thr Gly Ala Thr Gly Gly Cys Ala Gly Thr Gly Ala Ala Cys Gly
450 455 460
Ala Cys Ala Ala Ala Ala Thr Gly Gly Cys Gly Thr Cys Cys Thr Gly
465 470 475 480
Ala Ala Cys Ala Gly Thr Thr Gly Gly Ala Cys Thr Gly Ala Thr Cys
485 490 495
Ala Gly Gly Ala Cys Ala Gly Cys Ala Ala Ala Gly Ala Cys Ala Gly
500 505 510
Cys Ala Cys Cys Thr Ala Cys Ala Gly Cys Ala Thr Gly Ala Gly Cys
515 520 525
Ala Gly Cys Ala Cys Cys Cys Thr Cys Ala Cys Gly Thr Thr Gly Ala
530 535 540
Cys Cys Ala Ala Gly Gly Ala Cys Gly Ala Gly Thr Ala Thr Gly Ala
545 550 555 560
Ala Cys Gly Ala Cys Ala Cys Ala Ala Cys Ala Gly Cys Thr Ala Thr
565 570 575
Ala Cys Cys Thr Gly Thr Gly Ala Gly Gly Cys Cys Ala Cys Thr Cys
580 585 590
Ala Cys Ala Ala Gly Ala Cys Ala Thr Cys Ala Ala Cys Thr Thr Cys
595 600 605
Ala Cys Cys Cys Ala Thr Thr Gly Thr Cys Ala Ala Gly Ala Gly Cys
610 615 620
Thr Thr Cys Ala Ala Cys Ala Gly Gly Ala Ala Thr Gly Ala Gly Thr
625 630 635 640
Gly Thr
<210> 46
<211> 639
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Cys Ala Ala Ala Thr Thr Gly Thr Thr Cys Thr Cys Ala Cys Cys Cys
1 5 10 15
Ala Gly Thr Cys Thr Cys Cys Ala Gly Cys Ala Ala Thr Cys Ala Thr
20 25 30
Gly Thr Cys Thr Gly Cys Ala Thr Cys Thr Cys Cys Ala Gly Gly Gly
35 40 45
Gly Ala Gly Ala Ala Gly Gly Thr Cys Ala Cys Cys Ala Thr Gly Ala
50 55 60
Cys Cys Thr Gly Cys Ala Gly Thr Gly Cys Cys Ala Gly Cys Thr Cys
65 70 75 80
Ala Ala Gly Thr Gly Thr Ala Ala Gly Thr Thr Ala Cys Ala Thr Gly
85 90 95
Thr Ala Cys Thr Gly Gly Thr Ala Thr Cys Ala Gly Cys Ala Gly Ala
100 105 110
Ala Gly Gly Cys Ala Gly Gly Ala Thr Cys Cys Thr Cys Cys Cys Cys
115 120 125
Cys Ala Gly Ala Cys Thr Cys Cys Thr Gly Ala Thr Thr Thr Ala Thr
130 135 140
Gly Ala Cys Ala Cys Ala Thr Cys Cys Ala Ala Cys Cys Thr Gly Gly
145 150 155 160
Cys Thr Thr Cys Thr Gly Gly Ala Gly Thr Cys Cys Cys Thr Gly Thr
165 170 175
Thr Cys Gly Cys Thr Thr Cys Ala Gly Thr Gly Gly Cys Ala Gly Thr
180 185 190
Gly Gly Gly Thr Cys Thr Gly Gly Gly Ala Cys Cys Thr Cys Thr Thr
195 200 205
Ala Cys Thr Cys Thr Cys Thr Cys Ala Cys Ala Ala Thr Cys Ala Gly
210 215 220
Cys Cys Gly Ala Ala Thr Gly Gly Ala Gly Gly Cys Thr Gly Ala Ala
225 230 235 240
Gly Ala Thr Gly Cys Thr Gly Cys Cys Ala Cys Thr Thr Ala Thr Thr
245 250 255
Ala Cys Thr Gly Cys Cys Ala Gly Cys Ala Gly Thr Gly Gly Ala Gly
260 265 270
Thr Ala Gly Gly Thr Ala Cys Cys Cys Gly Cys Thr Cys Ala Cys Thr
275 280 285
Thr Thr Cys Gly Gly Thr Gly Gly Thr Gly Gly Gly Ala Cys Cys Ala
290 295 300
Ala Gly Gly Thr Gly Gly Ala Gly Cys Thr Gly Ala Ala Ala Cys Gly
305 310 315 320
Gly Gly Cys Thr Gly Ala Thr Gly Cys Thr Gly Cys Ala Cys Cys Ala
325 330 335
Ala Cys Thr Gly Thr Ala Thr Cys Cys Ala Thr Cys Thr Thr Cys Cys
340 345 350
Cys Ala Cys Cys Ala Thr Cys Cys Ala Gly Thr Gly Ala Gly Cys Ala
355 360 365
Gly Thr Thr Ala Ala Cys Ala Thr Cys Thr Gly Gly Ala Gly Gly Thr
370 375 380
Gly Cys Cys Thr Cys Ala Gly Thr Cys Gly Thr Gly Thr Gly Cys Thr
385 390 395 400
Thr Cys Thr Thr Gly Ala Ala Cys Ala Ala Cys Thr Thr Cys Thr Ala
405 410 415
Cys Cys Cys Cys Ala Ala Ala Gly Ala Cys Ala Thr Cys Ala Ala Thr
420 425 430
Gly Thr Cys Ala Ala Gly Thr Gly Gly Ala Ala Gly Ala Thr Thr Gly
435 440 445
Ala Thr Gly Gly Cys Ala Gly Thr Gly Ala Ala Cys Gly Ala Cys Ala
450 455 460
Ala Ala Ala Thr Gly Gly Cys Gly Thr Cys Cys Thr Gly Ala Ala Cys
465 470 475 480
Ala Gly Thr Thr Gly Gly Ala Cys Thr Gly Ala Thr Cys Ala Gly Gly
485 490 495
Ala Cys Ala Gly Cys Ala Ala Ala Gly Ala Cys Ala Gly Cys Ala Cys
500 505 510
Cys Thr Ala Cys Ala Gly Cys Ala Thr Gly Ala Gly Cys Ala Gly Cys
515 520 525
Ala Cys Cys Cys Thr Cys Ala Cys Gly Thr Thr Gly Ala Cys Cys Ala
530 535 540
Ala Gly Gly Ala Cys Gly Ala Gly Thr Ala Thr Gly Ala Ala Cys Gly
545 550 555 560
Ala Cys Ala Thr Ala Ala Cys Ala Gly Cys Thr Ala Thr Ala Cys Cys
565 570 575
Thr Gly Thr Gly Ala Gly Gly Cys Cys Ala Cys Thr Cys Ala Cys Ala
580 585 590
Ala Gly Ala Cys Ala Thr Cys Ala Ala Cys Thr Thr Cys Ala Cys Cys
595 600 605
Cys Ala Thr Thr Gly Thr Cys Ala Ala Gly Ala Gly Cys Thr Thr Cys
610 615 620
Ala Ala Cys Ala Gly Gly Ala Ala Thr Gly Ala Gly Thr Gly Thr
625 630 635
<210> 47
<211> 642
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Gly Ala Thr Ala Thr Cys Cys Ala Gly Ala Thr Gly Ala Cys Thr Cys
1 5 10 15
Ala Gly Thr Cys Thr Cys Cys Ala Gly Cys Cys Thr Cys Ala Cys Thr
20 25 30
Gly Thr Cys Thr Gly Cys Ala Thr Cys Thr Gly Thr Gly Gly Gly Ala
35 40 45
Gly Ala Ala Ala Cys Thr Gly Thr Cys Ala Cys Cys Ala Thr Cys Ala
50 55 60
Cys Ala Thr Gly Thr Gly Gly Ala Gly Cys Ala Ala Gly Thr Gly Ala
65 70 75 80
Gly Ala Ala Thr Ala Thr Thr Thr Ala Cys Gly Gly Thr Gly Cys Thr
85 90 95
Thr Thr Ala Ala Ala Thr Thr Gly Gly Thr Ala Thr Cys Ala Gly Cys
100 105 110
Thr Gly Ala Ala Ala Cys Ala Gly Gly Gly Ala Ala Ala Ala Thr Cys
115 120 125
Thr Cys Cys Thr Cys Ala Gly Cys Thr Cys Cys Thr Gly Ala Thr Cys
130 135 140
Thr Ala Thr Gly Gly Thr Gly Cys Gly Ala Cys Cys Ala Ala Cys Thr
145 150 155 160
Thr Gly Gly Cys Ala Gly Ala Thr Gly Gly Cys Thr Thr Gly Thr Cys
165 170 175
Ala Thr Cys Gly Ala Gly Gly Thr Thr Cys Ala Gly Ala Gly Gly Cys
180 185 190
Ala Gly Thr Gly Gly Ala Thr Cys Thr Gly Gly Thr Ala Gly Ala Cys
195 200 205
Ala Gly Thr Ala Thr Thr Cys Thr Cys Thr Cys Ala Ala Gly Ala Thr
210 215 220
Cys Ala Gly Thr Ala Ala Cys Cys Thr Gly Cys Ala Thr Cys Cys Thr
225 230 235 240
Gly Ala Cys Gly Ala Thr Gly Cys Thr Gly Cys Ala Ala Thr Gly Thr
245 250 255
Ala Thr Thr Ala Thr Thr Gly Thr Cys Ala Ala Ala Ala Thr Gly Thr
260 265 270
Gly Thr Thr Ala Ala Ala Thr Ala Cys Thr Cys Cys Gly Cys Thr Cys
275 280 285
Ala Cys Gly Thr Thr Cys Gly Gly Thr Gly Cys Thr Gly Gly Gly Ala
290 295 300
Cys Cys Ala Ala Gly Cys Thr Gly Gly Ala Gly Cys Thr Gly Ala Ala
305 310 315 320
Ala Cys Gly Gly Gly Cys Thr Gly Ala Thr Gly Cys Thr Gly Cys Ala
325 330 335
Cys Cys Ala Ala Cys Thr Gly Thr Ala Thr Cys Cys Ala Thr Cys Thr
340 345 350
Thr Cys Cys Cys Ala Cys Cys Ala Thr Cys Cys Ala Gly Thr Gly Ala
355 360 365
Gly Cys Ala Gly Thr Thr Ala Ala Cys Ala Thr Cys Thr Gly Gly Ala
370 375 380
Gly Gly Thr Gly Cys Cys Thr Cys Ala Gly Thr Cys Gly Thr Gly Thr
385 390 395 400
Gly Cys Thr Thr Cys Thr Thr Gly Ala Ala Cys Ala Ala Cys Thr Thr
405 410 415
Cys Thr Ala Cys Cys Cys Cys Ala Ala Ala Gly Ala Cys Ala Thr Cys
420 425 430
Ala Ala Thr Gly Thr Cys Ala Ala Gly Thr Gly Gly Ala Ala Gly Ala
435 440 445
Thr Thr Gly Ala Thr Gly Gly Cys Ala Gly Thr Gly Ala Ala Cys Gly
450 455 460
Ala Cys Ala Ala Ala Ala Thr Gly Gly Cys Gly Thr Cys Cys Thr Gly
465 470 475 480
Ala Ala Cys Ala Gly Thr Thr Gly Gly Ala Cys Thr Gly Ala Thr Cys
485 490 495
Ala Gly Gly Ala Cys Ala Gly Cys Ala Ala Ala Gly Ala Cys Ala Gly
500 505 510
Cys Ala Cys Cys Thr Ala Cys Ala Gly Cys Ala Thr Gly Ala Gly Cys
515 520 525
Ala Gly Cys Ala Cys Cys Cys Thr Cys Ala Cys Gly Thr Thr Gly Ala
530 535 540
Cys Cys Ala Ala Gly Gly Ala Cys Gly Ala Gly Thr Ala Thr Gly Ala
545 550 555 560
Ala Cys Gly Ala Cys Ala Thr Ala Ala Cys Ala Gly Cys Thr Ala Thr
565 570 575
Ala Cys Cys Thr Gly Thr Gly Ala Gly Gly Cys Cys Ala Cys Thr Cys
580 585 590
Ala Cys Ala Ala Gly Ala Cys Ala Thr Cys Ala Ala Cys Thr Thr Cys
595 600 605
Ala Cys Cys Cys Ala Thr Thr Gly Thr Cys Ala Ala Gly Ala Gly Cys
610 615 620
Thr Thr Cys Ala Ala Cys Ala Gly Gly Ala Ala Thr Gly Ala Gly Thr
625 630 635 640
Gly Thr
<210> 48
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
caggtcacct tgaaggagtc tgggggaggc ttggtgcagc ctggggggtc tctgacactc 60
tcttgtgccg ccagcggata cactttttat tctaacgcta tgggctgggc ccgccaggct 120
ccaggaaagg ggctcgagtg ggtgagcgcg atttctcgtt cttctggttc tatcaactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
cttcaaatga acagcctgaa acctgaggac acggccgttt attactgtgc agcctctagt 300
cgtctcgtca gtctcgattc tactggtgct cgtacgtatg gttactgggg tcagggtact 360
caggtcaccg tctcctca 378
<210> 49
<211> 369
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
caggtgcagc tgcaggagtc tggaggaggc ttggtgcagg ctggagggtc tctgagactc 60
tcctgtgcag ccgatggatt caccttccat cgctactaca tgggctgggc ccgccaggct 120
ccaggaaagg ggctcgagtg ggtcagcgcg atttctgcta ttggtcgcac atactatgca 180
gactccgtga agggccgatt caccatctcc agagacaacg ccaagaacac ggtgtatctt 240
caaatgaaca gcctgaaacc tgaggacacg gccgtttatt actgtgcagc ccgtctggac 300
tcttctctgt actactcttc tcgtccgtac gactactggg gtcagggtac tcaggtcacc 360
gtctcctca 369
<210> 50
<211> 351
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
caggtgcagc tgcaggagtc tggaggaggc ttggtgcagg ctggagggtc tctgagactc 60
tcttgtgccg ccagcggata cacctattct agtaatgtta tgagctgggc ccgccaggct 120
ccagggaagg agctcgagtt ggtgagcgga tttttttacg atggaactat gcactatgca 180
aactccgtga agggccgatt catcatctct agagacaccg gcaagcagac tatgtatctt 240
caaatgaaca gcctgaaacc tgaggacacg gccgtttatt actgcgcagc cgactacggt 300
agtagcccat ttgcttactg gggtcagggt actcaggtca ccgtctcctc a 351
<210> 51
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
caggtgcagc tgcaggagtc tgggggaggc ttggtgcgac ctggggggtc tctgagactc 60
tcctgtgcag cctctgcatc gaccttcagt tattactaca tgagctgggt ccgccaggct 120
ccaggaaagg ggctcgagtg ggtctcagcg atttctcgtt ctgactcttc tacttactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
cttcaaatga acagcctgaa acctgaggac acggccgttt attactgtgc agccctggct 300
tacaacaact cttactctcg ttactacgac tactggggtc agggtactca ggtcaccgtc 360
tcctca 366
<210> 52
<211> 372
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
caggtgcagc tgcaggagtc tggaggaggc ttggtgcagg ctggagggtc tctgagactc 60
tcttgtgccg ccagcggatc tatctcttct agttatgtta tgagctggtt ccgccaggct 120
ccagggaagg agcgtgagtt cgtagcagcg atttcttggt ctggttcttc tatctactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
cttcaaatga acagcctgaa acctgaggac acggccgttt attactgtgc agccggttct 300
tacctggact acccgtctcc gaactactct tacgactact ggggtcaggg tactcaggtc 360
accgtctcct ca 372
<210> 53
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
caggtgcagc tgcaggagtc tggaggaggc ttggtgcagg ctggagggtc tctgagactc 60
tcttgtgccg ccagcggatc tacttcttct acgtatacta tgacttggtt ccgccaggct 120
ccagggaagg agcgtgagtt cgtagcagcc atcagcaatc agaacagtaa cggtacctat 180
tacgccgaca gcgtgaaggg ccgattcacc atctccagag acaacgccaa gaacacggtg 240
tatcttcaaa tgaacagcct gaaacctgag gacacggccg tttattactg tgcagccggt 300
tcttacctgg actacccgtc tccgaacccg tactacgact actggggtca gggtactcag 360
gtcaccgtct cctca 375
<210> 54
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
caggtcacct tgaaggagtc tgggggaggc ttggtgcagc ctggggggtc tctgacactc 60
tcctgtgcag ccgatggatt caccttttat cgtaacgtta tgggctgggc ccgccaggct 120
ccagggaagg agctcgaatg ggtgtccgag attaatagtg gtgctggaga gacattctat 180
gcagactccg tgaagggccg attcatcatc tctagagaca ccggcaagca gactatgtat 240
ctgcaaatga acagcctgaa gcctgaggac acggccgttt attactgtgc aagacaagag 300
tatggtaatg ctatggacta ctggggtcaa ggaacccaag tcaccgtctc ctca 354
<210> 55
<211> 372
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
caggtgcagc tgcaggagtc tggaggaggc ttggtgcagg ctggagggtc tctgagactc 60
tcttgtgccg ccagcggata cactttctct agtaacacta tgggctggtt ccgccaggct 120
ccagggaagg agcgtgagtt cgtagcagcc atcagcagtt ggggtggtga tagttactac 180
gccgacagcg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
cttcaaatga acagcctgaa acctgaggac acggccgttt attactgtgc agccgactct 300
tacctgctgt acggtcgtga cccgaactac tacgactact ggggtcaggg tactcaggtc 360
accgtctcct ca 372
<210> 56
<211> 372
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
caggtgcagc tgcaggagtc tggaggaggc ttggtgcagg ctggagggtc tctgagactc 60
tcttgtgccg ccagcggata caccaacagt ggctcgacta tgagctggtt ccgccaggct 120
ccagggaagg agcgtgagtt cgtagcagcc atcagcaata cgagtggtga taagtactac 180
gccgacagcg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
cttcaaatga acagcctgaa acctgaggac acggccgttt attactgtgc agccggttct 300
tacctggact acccgtctaa ctactactct tacgactact ggggtcaggg tactcaggtc 360
accgtctcct ca 372
<210> 57
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
gatgtccagc tgcaggcgtc tggaggagga ttggtgcagg ctgggggctc tctgagactc 60
tcctgtgtag cctctggtgt taccttaact ccatatacca tgggctggtt ccgccaggct 120
ccagggaagg agcgtgagtt tgtagcagct acaattaggt tcaacggtaa cacatactat 180
gaagactccg tgaagggccg attcaccatc tccagagaca acgccaagga cacggtgtat 240
ctgcaaatga acaacctgaa acctgaggac acggccgttt attactgtgc agccgataat 300
ctgccatggt tcacggaatg ggcgaccctt aaatatgcct actggggaca ggggacccag 360
gtcaccgtct cctca 375
<210> 58
<211> 360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
gaggtgcagc tgcaggcgtc tggaggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactttggac tattatgcca tagcctggtt ccgccaggcc 120
ccagggaagg agcgtgaggg ggtctcgcgt attagtagta gtgatggtag cactgtgtat 180
gcagactccg tggcgggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgacgac acagccgttt attactgtgc agcaggcttg 300
gacggagacg gggtcgagta tgactactgg ggccagggga cccaggtcac cgtctcctca 360
<210> 59
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 59
gaggtgcagc tgcaggcgtc tgggggaggc atggtgcagc ctggggagtc tctgagactc 60
tcctgtgcag cctctggaag catcttcagt cgtaatacca tgagctggta ccgccagact 120
ccagggaatc agcgcgagtt ggtcgcaact gatactaatg gtggaagcac agtctatggc 180
gactccgtga agggccgatt caccatctcc agagacaact ttaagcacac ggtgtatctc 240
caaatgaaca gcctgaaacc tgaggacacg gccgtctatt actgtaatgc agaggccctc 300
gggagttact acgatctgag gtatctcgaa gtttggggcc agggcaccca ggtcaccgtc 360
tcctca 366
<210> 60
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 60
gatgtgcagc tgcaggcgtc tgggggaggc ttggtgcaac ctgggggttc tctgaggctc 60
tcctgtgcag cctctggatt caccttcagt agctatggta tgagctgggt ccgccaggct 120
ccaggaaagg ggctcgagtg ggtctcagtt attaatactg gcgatggtag cacggattat 180
gcagaccccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacgctgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgtgt attactgtgc aaagccgggc 300
cccggtaccc tagtagctgg ccccaggagg ttatatgagt ttgactactt gggccagggg 360
acccaggtca ccgtctcctc a 381
<210> 61
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
gatgtgcagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactttgaat tcatataaca tgggctggtt ccgccaggcc 120
ccaggggagg agcgcgaggg ggtggcatgt attactatgc atgatgaaag cacatactat 180
gcagactccg tggagggccg atccaccatc tccagagaca acaccaagaa cacgatgtat 240
ctgcaaatga acagcctgaa acctgaggac acagccgttt attactgtgc agcaggttgc 300
cgtagctact tcataacccc gggagatgac tactggggcc aggggaccca ggtcaccgtc 360
tcctca 366
<210> 62
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
gatgtgcagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggaatc tctgagactc 60
tcctgtgcag cctctggatt cactttggat tataccgcca taggctggtt ccgccaggcc 120
ccagggaagg agcgcgaggg ggtctcatgc attgttagta gtgatggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca atgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgttt attggtgtgc gacgagtggg 300
tgcgaagact acgggtatga ctggggccag gggacccagg tcaccgtctc ctca 354
<210> 63
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
gaggtccagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactatggat tattatgcca taggctggtt ccgccaggcc 120
ccagggaagg agcgagaggc gatctcatgt attagtagta gtgatggttt tacatacgat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acagccgttt attactgtgc agcgggggcc 300
ttagattggg gtattcgggc gatgtgtgtg atggccaaat atgactactg gggccagggg 360
acccaggtca ccgtctcctc a 381
<210> 64
<211> 372
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
gatgtccagc tgcaggcgtc tgggggagga ttggtgcagg ctgggggctc tctgagactc 60
tcctgtgcag cctctggacg caccttcagt agctatgccg tgggctggtt ccgccaggct 120
ccagggaagg agcgtgagct tgtagcagcc attaattgga gttctatcag cccatatgca 180
tactatgcag actccatgaa gggccgattc accatctcca gagacaacgc caagaacacg 240
gtgtatctgc aaatgaacag cctgaaacct gaggacacgg ccgtgtatag ctgtgcagcg 300
aatagcgact ataatgatgt atgggacgct agggactact ggggcaaagg gacccaggtc 360
accgtctcct ca 372
<210> 65
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 65
gatgtgcagc tgcaggcgtc tgggggagga ttggtgcagg ctgggggctc tctgagactc 60
tcctgtgcag ccgctgcacg caccttcagt cgctatgccg tgggctggtt ccgccaggct 120
ccaggaaagg agcgtgatct tgtagcagct attagctgga gtggcggtag tccatactat 180
tcagactcca tgaagggccg attcaccatc tccagagaca acaccaagaa cacggtgtat 240
ctgcaaatca acagcctgaa acctgaggac acggccgttt attactgtgc agcgaatagc 300
gactataatg atgtatggga cgacagggac tactggggca aggggaccca ggtcaccgtc 360
tcctca 366
<210> 66
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 66
gatgtgcagc tgcaggcgtc tgggggaggc tcggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactttggat tattataaca tgggctggtt ccgccaggcc 120
ccaggggagg agcgcgaggg ggtcgcatgt attagtagta atgatggtag cacatactat 180
gcagactccg tggcggggcg attcaccatc tccagagaca ataccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgttt attcctgtgc gactggttgc 300
ggcactgggg gttatgacta ctggggccag gggacccagg tcaccgtctc ctca 354
<210> 67
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 67
gaggtccagc tgcaggcgtc tgggggagga ttggtgcagg ctggggggtc tctgagactc 60
tcctgtgcag cctctggacg caccttcagt agctatgcca tgggctggtt ccgccaggcc 120
ccagggaagg agcccgaggg cgtctcatgt attgttagca gtgatggtag cacatactat 180
gcagactccg tggagggccg tttcaccatc tccagagaca atgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgttt attggtgtgc gacgagttgt 300
ggggaagacg ggtataacta ctggggccag gggacccagg tcaccgtctc ctca 354
<210> 68
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 68
gatgtgcagc tgcaggcgtc tggaggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggaag catcttcagt atgcctgcca tgggctggta ccgccaggct 120
ccagggaagc agcgcgagtt ggtcgcgtca tttcgtggtg gtagtacaaa ctatgcagac 180
ttcgttcagg gccgattcac catctccaga gacgacgcca agaacgcggt gtatctgcaa 240
atgaacagcc tgaaacctgg ggacacggcc gtctattact gtcatgcgaa ggccctcggg 300
agttactacg atctgacgta tctcgaagtt tggggccagg gcacccaggt caccgtctcc 360
tca 363
<210> 69
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
gatgtgcagc tgcaggcgtc tggaggagga ttggtgcagg ctgggggctc tctgagactc 60
tcctgtgcag cctctggacg caccttcagt acctatgcca tcggctggtt ccgccaggct 120
ccagggaagg agcgtgagct tgtagcagct attagctgga gtggtggtag cccgtactat 180
gcagacccca tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatag acagtctgaa acctgaggac acggccgttt attactgtgc agcgaatagc 300
gactataatg atgtatggga cgaacgggac tactggggca aagggaccca ggtcaccgtc 360
tcctca 366
<210> 70
<211> 357
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
gatgtccagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggaac caccttcagt atcaatgcca tgggctggta ccgccaggct 120
ccagggaagc aggtcgagtt ggtcgcaaat attagtagtg gtggtagccc aaactatgca 180
gactccgtga agggccgatt caccatctcc agagacaacg ccaagaacac ggtgtatctg 240
caaatgaaca gcctgaaacc tgaggacacg gccgtctatt attgtcatgt tacctacggg 300
ccctcctcgt ggggcatgga ctactggggc aaagggaccc aggtcaccgt ctcctca 357
<210> 71
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
gaggtgcagc tgcaggcgtc tgggggaggc atggtgcagc ctggggagtc tctgagactc 60
tcctgtgcag cctctggaag catcttcagt atcaagacca tgggctggta ccgccagact 120
ccagggaagc agcgcgagtt ggtcgcaacg catactaatg gtggtagcac aaactatggc 180
gactccgtga agggccgatt caccatctcc agagacaact ttaaacacgc ggtgtatctc 240
caaatgaaca gcctgaaacc tgaggacacg gccgtctatt actgtaatgc agaggccctc 300
gggagttact acgatctgaa gtatctcgaa gtttggggcc agggcaccca ggtcaccgtc 360
tcctca 366
<210> 72
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
gatgtgcagc tgcaggcgtc tgggggagga ttggtgcagg ctgggggctc tctgagactc 60
tcctgtgcag cctctggacg caccgccagt agctatgcca tgggctggtt ccgccaggct 120
ccagggaagg agcgtgagct actagcagct attagctgga gtggtggtag cccatactat 180
gcagactcca tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgttt attactgtgc agcgaatagc 300
gactataatg atgtatggga cggcagggac tactggggca aagggaccca ggtcaccgtc 360
tcctca 366
<210> 73
<211> 360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
cagtcggtga aggagtccga gggtcgcctg gtcacgcctg ggacacccct gacactcacc 60
tgcacagtct ctggaatcga cctcagtatc tatccagtga gttgggtccg ccaggctcca 120
gggaagggcc tggaatacat cggaaccatg actgctagtg gtaacacata ctacgcgaac 180
tgggcaaatg gtcgattcac catctccaaa acctcgacca cggtggatct gaaaatgacc 240
agtccgacaa ccgaggacac ggccacctat ttccgcgcca gagggggctc tactggtgct 300
acttattcgg ctcgtgcttt tgatccctgg ggcccaggca ccctggtcac cgtctcctca 360
<210> 74
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
gaggtgcagc tgcaggcgtc tggaggagga ttggtgcagg ctgggggctc tctgagactc 60
gcctgtgaag cctctggact ttccttcagt gtcaatacca taggctggta ccgccaggcc 120
ccagggaagg agcgcgaggg ggtctcatgt cttgttagta gtgatggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca atgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgttt attggtgtgc gaggagttgt 300
ggggaagacg ggtatgacta ctggggccag gggacccagg tcaccgtctc ctca 354
<210> 75
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
gatgtgcagc tgcaggcgtc tggaggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cgtctagatt cactttggat tattatgccg taggctggtt ccgccgggcc 120
ccagggaagg agcccgaggc ggtctcatgt attactaata gtgatggtag cacctactat 180
gcagactccg tgaagggccg attcaccatc ttcagagaca atgccaagaa cacggtgtat 240
ctgcaaatga acagccttaa acctgaggac acggccgttt attactgtgc gccgactgga 300
gaggacggga tggactgtta caggggccag gggacccagg tcaccgtctc ctca 354
<210> 76
<211> 360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
gaggtccagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactttggat tattacaaca tgggctggtt ccgccagacc 120
ccggggcagg agcgtgaggg ggtcgcatgt attagtagtg gtgatggtag cacatactat 180
gcagactccg tggagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acagccgtat attactgtgc agtaggttgc 300
agccctttcg gcataccgga tgactactgg ggccagggga cccaggtcac cgtctcctca 360
<210> 77
<211> 393
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
gatgtccagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt caccttcagt agctactgga tgtattgggt ccgtcaggct 120
ccagggaagg ggctcgagtg ggtctcaggt attaatactg atagtggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagaaaca acgccaagaa cacgctatat 240
ctccaaatga acagtctgaa atctgaggac acggccgtgt attactgtgc aaaaccggga 300
gccggtacgg gagccgatat ggtagttgct ggtccgattg gactgattca acttggttcc 360
cggggccagg ggacccaggt caccgtctcc tca 393
<210> 78
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
gatgtccagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cgtctggatt cactttggat tattataaca taggctggtt ccgccaggcc 120
ccagggaagg agcccgaggc ggtctcatgt attagtagtg gtgatggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca atgccaagaa cacggtgtat 240
ctgcaaatga acaggctgaa acctgaggac acggccgttt attactgtgc gacgacttgt 300
ggggaagacg ggtatgacta cgggggccag gggacccagg tcaccgtctc ctca 354
<210> 79
<211> 366
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
gatgtgcagc tgcaggcgtc tggaggaggc ttggtgcagg ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactttcgat gattatgcca taggctggtt ccgccaggcc 120
ccagggaagg agcgtgaggg ggtcttatgt attagtgata gtgatggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acagccgttt attattgtac agctggtacg 300
gtagcagctg agtgggctta cggcatggac tactggggca aagggaccca ggtcaccgtc 360
tcctca 366
<210> 80
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
gaggtgcagc tgcaggcgtc tgggggaggc ttggtgcagc cgggggggtc tctgagactc 60
tcctgcgcag cctctggatt cactttggat cgttatacta taggctggtt ccgccaggcc 120
ccaggagagg agcgtgaggg ggttgcatgc cttagtagta gtgatggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acagccgttt attattgtgc cccgggttgt 300
agcccgtacg gcataaacta ctggggcaaa gggacccagg tcaccgtctc ctca 354
<210> 81
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
gatgtgcagc tgcaggcgtc tgggggagga ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggaag catcggcagt atgtatgtca tgggctggta ccgccaggct 120
ccagggaagc agcgcgactg ggtcgcacgt attagtctta ttaatggtag cacaaattat 180
gcggactccg tgaagggccg attcaccatc tccagaggca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acggccgtgt attattgtaa tgccgacttc 300
cgaccttttg ggtttgagta ctggggccag gggagtcagg tcaccgtctc ctca 354
<210> 82
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
gaggtgcagc tgcaggcgtc tggaggagga ttggtgcagg ctgggggctc tctgagactc 60
tcctgtgtaa gctctggagg cgccttcagt agctatacca tgggctggtt ccgccaggct 120
ccagggaagg agcgtgagtt tgtagcagct gatattaggt ataacggtaa cacatactat 180
gaagactccg tgaagggccg attcaccatc tccagagata gcgccaagag cacggtgtat 240
ctgcaaatga acaacctgaa acctgaggac acggccgttt attactgtgc aggcgatgcc 300
cgcccatggt tcacggaatg ggcgacggtt aaatatacat actggggaca ggggacccag 360
gtcaccgtct cctca 375
<210> 83
<211> 353
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
gaggtgcagc tgcaggcgtc tggaggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt cactttggat atgtatgtta taagctggtt ccgccaggcc 120
ccagggaagg agcctgaggg ggtctcatgt attagtagta gtgatggtag cacatactat 180
gcagactccg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgcat 240
ctgcaaatga acagcctgaa acctgaggac acagccgttt attggtgtgc agcaacttgc 300
gggaccgacg ggtatgacta ctggggccag gggacccagg tcaccgtctc ctc 353
<210> 84
<211> 393
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
gatgtgcagc tgcaggcgtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggatt caccttcagt agctactgga tgtattgggt ccgtcaggct 120
ccagggaagg ggctcgagtg ggtctcaggt attaatagtg atagtggtag cacatactac 180
gcagactccg tgaagggccg attcaccatc tccagaaaca acgccaagaa cacgctatat 240
ctccaaatga acagtctgaa atctgaggac acggccgtgt attactgtgc aaaaccggga 300
gccggtgcgg gagccgatat ggtagtagct ggtccgattg gactgattca actcggttcc 360
cggggccagg ggacccaggt caccgtctcc tca 393
Claims (10)
1.一种抗人SEMA4D抗体,其特征在于,所述抗人SEMA4D抗体能特异性地识别人SEMA4D蛋白;
所述抗人SEMA4D抗体包括第一抗体或第二抗体;
所述第一抗体包括SEQ ID NO:1-SEQ ID NO:37所示氨基酸序列中的任意一种,所述第二抗体序列满足如下至少一条:重链氨基酸序列包括SEQ ID NO:38-SEQ ID NO:42所示氨基酸序列中的任意一种,轻链氨基酸序列包括SEQ ID NO:43-SEQ ID NO:47所示氨基酸序列中的任意一种。
2.根据权利要求1所述的抗人SEMA4D抗体,其特征在于,所述抗人SEMA4D抗体还包括:与所述第一抗体具有同源的氨基酸序列的第一同源抗体,或与所述第二抗体具有同源氨基酸序列的第二同源抗体,其中,
所述第一同源抗体的CDR1区氨基酸序列与所述第一抗体中相对应的CDR1区具有80%以上同源性;
所述第一同源抗体的CDR2区氨基酸序列与所述第一抗体中相对应的CDR2区具有70%以上同源性;
所述第一同源抗体的CDR3区氨基酸序列与所述第一抗体中相对应的CDR3区具有75%以上同源性;
所述第二同源抗体的CDR1区氨基酸序列与所述第二抗体中相对应的CDR1区具有80%以上同源性;
所述第二同源抗体的CDR2区氨基酸序列与所述第二抗体中相对应的CDR2区具有70%以上同源性;
所述所述第二同源抗体的CDR3区氨基酸序列与所述第二抗体中相对应的CDR3区具有75%以上同源性。
4.一种制备如1-3中任意一项所述的抗人SEMA4D抗体的方法,其特征在于,所述方法包括下列步骤:
取正常血清,或用SEMA4D重组蛋白进行多次免疫后得到的血清,分离得到淋巴细胞;
从所述淋巴细胞中提取RNA,进行cDNA的合成,得到cDNA片段;
以所述cDNA为模板,进行PCR扩增,得到基因片段;
将所述基因片段插入载体的DNA序列中,并去除未插入成功的载体,得到含所述基因的载体;
将含所述基因的所述载体感染感受态细胞,进行培养,检测抗人SEMA4D抗体的表达的程度。
5.一种编码如权利要求1-3中任意一项所述抗人SEMA4D抗体的基因序列,其特征在于,所述基因序列包括第一基因序列和第二基因序列;所述第一基因序列如SEQ ID NO:48-SEQID NO:77所示核苷酸序列中的任意一种;所述第二基因序列包括以下至少一条:重链基因如SEQ ID NO:78-SEQ ID NO:82所示的核苷酸序列中任意一种,轻链基因如SEQ ID NO:83-SEQ ID NO:87所示的核苷酸序列中任意一种。
6.一种含有权利要求5所述基因序列的载体。
7.一种转化融合细胞,其特征在于,所述转化融合细胞含有权利要求5所述基因序列,或含有权利要求6所述载体。
8.根据权利要求7所述的转化融合细胞,所述转化融合细胞包括哺乳细胞、酵母细胞、昆虫细胞和原核细胞中任意一种。
9.一种包含权利要求1-3中任意一项所述抗人SEMA4D抗体在诊断标志物及治疗靶点中的应用。
10.一种SEMA4D蛋白的检测方法,其特征在于,所述方法包括使用如权利要求1-3中任意一项所述的抗人SEMA4D抗体进行检测。
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Denomination of invention: Anti human SEMA4D antibody and its preparation method and application Granted publication date: 20230829 Pledgee: Guanggu Branch of Wuhan Rural Commercial Bank Co.,Ltd. Pledgor: CUSABIO BIOTECH Co.,Ltd. Registration number: Y2024980009781 |