CN110305220B - 一种癌靶向增强型抗肿瘤融合蛋白及制备方法及用途 - Google Patents

一种癌靶向增强型抗肿瘤融合蛋白及制备方法及用途 Download PDF

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CN110305220B
CN110305220B CN201810258777.6A CN201810258777A CN110305220B CN 110305220 B CN110305220 B CN 110305220B CN 201810258777 A CN201810258777 A CN 201810258777A CN 110305220 B CN110305220 B CN 110305220B
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孙嘉琳
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Nanjing Double Bond Targeted Drug Technology Co ltd
Sun Jialin
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Abstract

本发明公开了一种癌靶向增强型抗肿瘤融合蛋白及制备方法及用途，所述癌靶向增强型抗肿瘤融合蛋白包括：a)抗体和b)诱导并激活T细胞的融合蛋白。本发明的癌靶向增强型抗肿瘤融合蛋白中各组份间具有协同作用，产生叠加放大的效果，显示的生物活性远远超过单个分子；诱导并激活T细胞来杀死肿瘤，增强了T细胞分泌的细胞因子的作用，实验证明，本发明的癌靶向增强型抗肿瘤融合蛋白增强了干扰素‑γ(IFN‑γ)、颗粒酶的分泌数量，从而加速癌细胞凋亡，融合蛋白对于各种癌症和实体肿瘤有着很明显的杀伤作用。

Description

一种癌靶向增强型抗肿瘤融合蛋白及制备方法及用途
技术领域
本发明涉及一种融合蛋白，特别是涉及一种癌靶向增强型抗肿瘤融合蛋白及制备方法及用途。
背景技术
抗体技术已广泛应用于各种疾病的靶向治疗，特别是肿瘤治疗，减少了化学药物带来的副作用。已上市的抗体药物有：
针对HER2(ErbB2)靶点的Trastuzumab(Herceptin)抗体，它应用于乳腺癌的治疗(Proc Natl Acad Sci USA,89,4285-4289,1992；Clin Cancer Res,12,904-916,2006；Cancer Res,69,9330-9336,2009)；
针对CD20靶点的Rituximab(Rituxan)抗体，它应用于B细胞由来淋巴瘤的治疗(Cancer Res,68,2400-2408,2008；Cell Physiol Biochem,32,645-654,2013)；
针对EGFR靶点的Cetuximab(Erbitux)抗体，它应用于结肠癌的治疗(Clin CancerRes,1,1311-1318,1995；Clin Cancer Res,8,1253-1264,2002；Clin Cancer Res,21,4153-4164,2015；J Nucl Med,5,1239-1245,2015)
针对EGFR靶点的Panitumumab(Vectibix)抗体，它应用于结肠癌的治疗(MolCancer Ther,9,2297-2308,2010；Mol Cancer,11,47,2012；J Nucl Med,53,113-120,2012)；
针对VEGFR2靶点的Ramucirumab(Cyramza)抗体，它应用于胃癌和肺癌的治疗(JExp Clin Cancer Res,34,132,2015；Ther Clin Risk Manag,11,1123-1132,2015；OncoTargets Ther,9,1953-1960,2016)。
但是已上市的抗体药物大都不利用T细胞，也就是在抗体药物治疗肿瘤的过程中，T细胞资源还没有被利用。如何有效利用T细胞靶向杀死肿瘤仍然是极具挑战的难题。
T细胞的有效活化需要两个信号的参与,即需要抗原肽-MHC复合物与T细胞上的TCR-CD3结合提供第1信号，以及共刺激分子(Costimulatory molecules)介导的第2信号。最基本的共刺激信号由抗原递呈细胞表达的B7.1(CD80)、B7.2(CD86)分子与T细胞上表达的相应受体(CD28和CTLA-4)提供(Amj Respir Crit Care Med,162,5164-5168,2000；Immunol Cell Biol,77,304-311,1999；Clin Cancer Res,13,5271-5279,2007；TrendsImmunol,24,313-318,2003)。
B7.1表达在活化的DC、B细胞和单核细胞上，B7.2在B细胞、T细胞、DC和巨噬细胞上呈低水平表达，但B7.2可被诱导快速上调，而B7.1被诱导上调表达时间比B7.2晚。初始T细胞上的CD28与APC上的B7.1/B7.2结合，可以促进IL-2转录，同时使IL-2受体在T细胞上表达增加，从而促进T细胞增殖，还可以通过增加Bcl-xl的表达途径使T细胞免于凋亡。
有报道，B7.1-Fc或B7.2-Fc可以诱导并激活抗癌的T细胞，显示出抗肿瘤的生物活性(Cancer Res,59,4964-4972,1999；Cancer Res,59,2650-2656,1999；Clin Cancer Res,11,8492-8502,2005；J Immunol,172,1347-1354,2004；Cancer Res,62,5727-5735,2002)。
超抗原(Superantigen)也能引起细胞毒作用，它是一类特殊的抗原分子，主要是一些细菌的毒素和逆转录病毒基因的产物，不需要抗原提呈细胞的加工处理，而以完整的蛋白质形式直接与细胞膜上的MHC II类分子结合形成复合物，识别TCR的Vb片段，激活比普通抗原多得多的T细胞(包括CD4+，CD8+)，并释放大量细胞因子，对靶细胞产生强而有力的细胞毒作用。
超抗原与人类多种急、慢性疾病的发生有关，但在抗肿瘤研究中也发挥了独特的作用，尝试用它激活的T细胞来杀伤肿瘤，并取得了一定的成果，目前较有研究基础的超抗原主要是金黄色葡萄球菌肠毒素A、B、C等(Technol Cancer Res Treat,16,125-132,2017；Biosci Biotechnol Biochem,81,1741-1746,2017；Anticancer Res,25,3565-3573,2005；Biochem Biophys Res Commun,290,1336-1342,2002；Cancer Immunol Immunother,33,231-237,1991)。
上述B7.1、B7.2或超抗原，在抑制肿瘤方面其活性仍有待提高。
发明内容
本发明的目的是克服现有技术中的不足，提供癌靶向增强型抗肿瘤融合蛋白。
本发明的第二个目的是提供含有编码癌靶向增强型抗肿瘤融合蛋白的核苷酸序列重组载体。
本发明的第三个目的是提供含有重组载体的宿主细胞。
本发明的第四个目的是提供癌靶向增强型抗肿瘤融合蛋白的制备方法。
本发明的第五个目的是提供癌靶向增强型抗肿瘤融合蛋白的应用。
本发明的技术方案概述如下：
癌靶向增强型抗肿瘤融合蛋白，包括：a)抗体和b)诱导并激活T细胞的融合蛋白。
优选地，抗体为单链抗体形式的Trastuzumab、Rituximab、Cetuximab、Panitumumab或Ramucirumab；
Trastuzumab的氨基酸序列用SEQ ID NO.12所示；Rituximab的氨基酸序列用SEQID NO.18所示；Cetuximab的氨基酸序列用SEQ ID NO.24所示；Panitumumab的氨基酸序列用SEQ ID NO.30所示；Ramucirumab的氨基酸序列用SEQ ID NO.36所示。
诱导并激活T细胞的融合蛋白为诱导并激活T细胞的共刺激分子B7.1和诱导并激活T细胞的共刺激分子B7.2融合；或诱导并激活T细胞的共刺激分子B7.1、诱导并激活T细胞的共刺激分子B7.2和超抗原金黄色葡萄球菌肠毒素A融合；
诱导并激活T细胞的共刺激分子B7.1简称B7.1；诱导并激活T细胞的共刺激分子B7.2简称B7.2；超抗原金黄色葡萄球菌肠毒素A简称SEA；
B7.1-B7.2的氨基酸序列用SEQ ID NO.8所示；B7.1-B7.2-SEA的氨基酸序列用SEQID NO.10所示。
癌靶向增强型抗肿瘤融合蛋白优选为：Trastuzumab-B7.1-B7.2融合且氨基酸序列用SEQ ID No.14所示；Trastuzumab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID No.16所示；Rituximab-B7.1-B7.2融合且氨基酸序列用SEQ ID No.20所示；Rituximab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID No.22所示；Cetuximab-B7.1-B7.2融合且氨基酸序列用SEQ ID No.26所示；Cetuximab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID No.28所示；Panitumumab-B7.1-B7.2融合且氨基酸序列用SEQ ID No.32所示；Panitumumab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID No.34所示；Ramucirumab-B7.1-B7.2融合且氨基酸序列用SEQ ID No.38所示；Ramucirumab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ IDNo.40所示。
重组载体，含有编码权利要求3所述癌靶向增强型抗肿瘤融合蛋白的核苷酸序列；编码SEQ ID No.14的核苷酸序列用SEQ ID No.13所示；编码SEQ ID No.16的核苷酸序列用SEQ ID No.15所示；编码SEQ ID No.20的核苷酸序列用SEQ ID No.19所示；编码SEQ IDNo.22的核苷酸序列用SEQ ID No.21所示；编码SEQ ID No.26的核苷酸序列用SEQ IDNo.25所示；编码SEQ ID No.28的核苷酸序列用SEQ ID No.27所示；编码SEQ ID No.32的核苷酸序列用SEQ ID No.31所示；编码SEQ ID No.34的核苷酸序列用SEQ ID No.33所示；编码SEQ ID No.38的核苷酸序列用SEQ ID No.37所示；或编码SEQ ID No.40的核苷酸序列用SEQ ID No.39所示。
宿主细胞，含有上述重组载体。
癌靶向增强型抗肿瘤融合蛋白的制备方法，培养宿主细胞，收集癌靶向增强型抗肿瘤融合蛋白。
上述癌靶向增强型抗肿瘤融合蛋白在制备治疗癌症和抗实体肿瘤药物的应用。
本发明的优点：
本发明的癌靶向增强型抗肿瘤融合蛋白中各组份间具有协同作用，产生叠加放大的效果，显示的生物活性远远超过单个分子；诱导并激活T细胞来杀死肿瘤，增强了T细胞分泌的细胞因子的作用，实验证明，本发明的癌靶向增强型抗肿瘤融合蛋白增强了干扰素-γ(IFN-γ)、颗粒酶的分泌数量，从而加速癌细胞凋亡，融合蛋白对于各种癌症和实体肿瘤有着很明显的杀伤作用。
附图说明
图1表示各种蛋白抑制S180/HER2(ErbB-2)肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中右边曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Trastuzumab组，3是Trastuzumab-B7.1组，4是Trastuzumab-B7.2组，5是Trastuzumab-SEA组，6是Trastuzumab-B7.1-B7.2组、7是Trastuzumab-B7.1-B7.2-SEA组。
图2表示第11天杀死小鼠解剖后的S180/HER2(ErbB-2)肿瘤重量，横轴表示实验用小鼠的编组，1是生理盐水组，2是Trastuzumab组，3是Trastuzumab-B7.1组，4是Trastuzumab-B7.2组，5是Trastuzumab-SEA组，6是Trastuzumab-B7.1-B7.2组、7是Trastuzumab-B7.1-B7.2-SEA组。
图3表示各种蛋白抑制S180/CD20肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中右边曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Rituximab组，3是Rituximab-B7.1组，4是Rituximab-B7.2组，5是Rituximab-SEA组，6是Rituximab-B7.1-B7.2组，7是Rituximab-B7.1-B7.2-SEA组。
图4表示第11天杀死小鼠解剖后的S180/CD20肿瘤重量，横轴表示实验用小鼠的编组，1是生理盐水组，2是Rituximab组，3是Rituximab-B7.1组，4是Rituximab-B7.2组，5是Rituximab-SEA组，6是Rituximab-B7.1-B7.2组，7是Rituximab-B7.1-B7.2-SEA组。
图5利用常规免疫组化检测携带S180/HER2(ErbB-2)肿瘤组织内的T细胞，棕色小点部分就是T细胞，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
图6利用常规免疫组化检测携带S180/CD20肿瘤组织内的T细胞，棕色小点部分就是T细胞，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
图7利用常规免疫组化检测携带S180/HER2(ErbB-2)肿瘤组织内的干扰素-γ(IFN-γ)，棕色部分就是T细胞分泌的IFN-γ，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
图8利用常规免疫组化检测携带S180/CD20肿瘤组织内的干扰素-γ(IFN-γ)，棕色部分就是T细胞分泌的IFN-γ，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
图9利用常规免疫组化检测颗粒酶(Granzyme B)，棕色部分就是T细胞分泌的颗粒酶，，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
图10利用常规免疫组化检测颗粒酶(Granzyme B)，棕色部分就是T细胞分泌的颗粒酶，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
图11利用常规免疫组化检测染色体断裂，棕色部分就是发生染色体断裂的肿瘤细胞，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
图12利用常规免疫组化检测染色体断裂，棕色部分就是发生染色体断裂的肿瘤细胞，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
图13表示抗体抑制肿瘤生长和第9天杀死荷瘤小鼠解剖后的肿瘤重量。
A表示各种蛋白抑制A431肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中上面曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Cetuximab组，3是Cetuximab-B7.1组，4是Cetuximab-B7.2组，5是Cetuximab-SEA组，6是Cetuximab-B7.1-B7.2组，7是Cetuximab-B7.1-B7.2-SEA组。
B表示第11天杀死小鼠解剖后的A431肿瘤重量，横轴表示的是实验用小鼠的编组，1是生理盐水组，2是Cetuximab组，3是Cetuximab-B7.1组，4是Cetuximab-B7.2组，5是Cetuximab-SEA组，6是Cetuximab-B7.1-B7.2组，7是Cetuximab-B7.1-B7.2-SEA组。
图14表示抗体抑制肿瘤生长和第9天杀死荷瘤小鼠解剖后的肿瘤重量。
A表示各种蛋白抑制A431肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中上面曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Panitumumab组，3是Panitumumab-B7.1组，4是Panitumumab-B7.2组，5是Panitumumab-SEA组，6是Panitumumab-B7.1-B7.2组，7是Panitumumab-B7.1-B7.2-SEA组。
B表示第9天杀死小鼠解剖后的A431肿瘤重量，横轴表示的是实验用小鼠的编组，1是生理盐水组，2是Panitumumab组，3是Panitumumab-B7.1组，4是Panitumumab-B7.2组，5是Panitumumab-SEA组，6是Panitumumab-B7.1-B7.2组，7是Panitumumab-B7.1-B7.2-SEA组。
图15表示抗体抑制肿瘤生长和第9天杀死荷瘤小鼠解剖后的肿瘤重量。
A表示各种蛋白抑制HCC4006肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中上面曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Ramucirumab组，3是Ramucirumab-B7.1组，4是Ramucirumab-B7.2组，5是Ramucirumab-SEA组，6是Ramucirumab-B7.1-B7.2，7是Ramucirumab-B7.1-B7.2-SEA组。
B表示第9天杀死小鼠解剖后的HCC4006肿瘤重量，横轴表示的是实验用小鼠的编组，1是生理盐水组，2是Ramucirumab组，3是Ramucirumab-B7.1组，4是Ramucirumab-B7.2组，5是Ramucirumab-SEA组，6是Ramucirumab-B7.1-B7.2，7是Ramucirumab-B7.1-B7.2-SEA组。
具体实施方式
下面通过具体实施例对本发明作进一步的说明。
建立小鼠荷瘤模型，检查融合蛋白杀伤肿瘤的效果和检测融合蛋白的诱导并激活T细胞生物活性的能力。
构建融合蛋白所需要的各种蛋白的基因序列确定如下：
B7.1基因信息参考B7.1论文(J Exp Med,174,625-631,1991)和GenBank数据库(NM_005191)，这里所采用的序列是它的细胞外片断，(核苷酸序列为SEQ ID No.1所示，氨基酸序列为SEQ ID No.2所示)。
B7.2基因信息参考B7.2的GenBank数据库(NM_175862和L25259)，这里所采用的序列是它的细胞外片断，(核苷酸序列为SEQ ID No.3所示，氨基酸序列为SEQ ID No.4所示)。
SEA基因信息参考SEA论文(J Biol Chem,262,7006-7013,1987；J Bacteriol,170,34-41,1988)和GenBank数据库(M18970)(核苷酸序列为SEQ ID No.5所示，氨基酸序列为SEQ ID No.6所示)。
B7.1-B7.2融合蛋白的核苷酸序列为SEQ ID No.7所示，氨基酸序列为SEQ IDNo.8所示，B7.1与B7.2是通过连接肽1(核苷酸序列为SEQ ID No.41所示，氨基酸序列为SEQID No.42所示)连接。
B7.1-B7.2-SEA融合蛋白的核苷酸序列为SEQ ID No.9所示，氨基酸序列为SEQ IDNo.10所示，B7.1-B7.2与SEA也通过连接肽1连接。
下述所有抗体形式采用抗体轻链可变区(VL)和重链可变区(VH)来构建单链抗体，轻链可变区(VL)和重链可变区(VH)之间用一个连接肽2(连接肽2的核苷酸序列为SEQ IDNo.43所示，氨基酸序列为SEQ ID No.44所示)来连接。
Trastuzumab(Herceptin)基因信息参考抗体论文(Proc Natl Acad Sci USA,89,4285-4289,1992)和GenBank数据库(KM016462.1、AY513484.1和AY513485.1)。这里采用抗体轻链可变区(VL)和重链可变区(VH)的序列(Trastuzumab单链抗体核苷酸序列为SEQ IDNo.11所示，氨基酸序列为SEQ ID No.12所示)。
Rituximab(Rituxan)基因信息参考GenBank数据库(KP783167.1)和PDB数据库(2OSL_A和2OSL_B)。这里采用抗体轻链可变区(VL)和重链可变区(VH)的序列(Rituximab单链抗体核苷酸序列为SEQ ID No.17所示，氨基酸序列为SEQ ID No.18所示)。
Cetuximab(Erbitux)基因信息参考GenBank数据库(KX138561.1和KX138562.1)和PDB数据库(5T1M_A和5T1M_B)。这里采用抗体轻链可变区(VL)和重链可变区(VH)的序列(Cetuximab单链抗体核苷酸序列为SEQ ID No.23所示，氨基酸序列为SEQ ID No.24所示)。
Panitumumab(Vectibix)基因信息参考PDB数据库(5SX5_L和5SX5_H)，根据这2个数据确定抗体的轻链和重链的氨基酸序列，然后通过BLAST对比寻找类似的轻链基因(GenBank:AY043104.1)和重链基因(GenBank:AY671470.1)，再调整或修改有关位置上的碱基，最后的核苷酸序列与PDB数据库(5SX5_L和5SX5_H)的氨基酸序列对应。这里采用抗体轻链可变区(VL)和重链可变区(VH)的序列(Panitumumab单链抗体核苷酸序列为SEQ IDNo.29所示，氨基酸序列为SEQ ID No.30所示)。
Ramucirumab(Cyramza)基因信息参考PDB数据库(3S34_L、3S34_H、3S37_L和3S37_H)，根据这些数据确定抗体的轻链和重链的氨基酸序列，然后通过BLAST对比寻找类似的轻链基因(GenBank:X97555.1)和重链基因(GenBank:EF177948.1)，再调整或修改有关位置上的碱基，最后的核苷酸序列与PDB数据库(3S34_L、3S34_H、3S37_L和3S37_H)的氨基酸序列对应。这里采用抗体轻链可变区(VL)和重链可变区(VH)的序列(Ramucirumab单链抗体核苷酸序列为SEQ ID No.35所示，氨基酸序列为SEQ ID No.36所示)。
实施例1构建表达各种蛋白的载体
所采用的表达质粒是pET22b(Novagen公司)，含有各种限制酶切位点(BamHI-EcoRI-SacI-SalI-HindIII-NotI-XhoI)供编码外源蛋白的DNA片断的插入，以及在表达外源蛋白的C端有His-tag(含有6个His)供纯化蛋白用。
DNA片断由TAKARA公司和上海生工生物工程公司合成。
片断Trastuzumab单链抗体的核酸序列由SEQ ID No.11所示，VL和VH之间由连接肽2(SEQ ID No.43)连接。
片断Trastuzumab单链抗体-B7.1融合蛋白是由SEQ ID No.11所示的Trastuzumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.1所示的B7.1核酸序列。
片断Trastuzumab单链抗体-B7.2融合蛋白是由SEQ ID No.11所示的Trastuzumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.3所示的B7.2核酸序列。
片断Trastuzumab单链抗体-SEA融合蛋白是由SEQ ID No.11所示的Trastuzumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.5所示的SEA核酸序列。
片断Trastuzumab单链抗体-B7.1-B7.2融合蛋白是由SEQ ID No.11所示的Trastuzumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.7所示的B7.1-B7.2融合蛋白核酸序列，Trastuzumab单链抗体-B7.1-B7.2融合蛋白的核酸序列由SEQ ID No.13所示。
片断Trastuzumab单链抗体-B7.1-B7.2-SEA融合蛋白是由SEQ ID No.11所示的Trastuzumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.9所示的B7.1-B7.2-SEA融合蛋白核酸序列，Trastuzumab单链抗体-B7.1-B7.2-SEA融合蛋白的核酸序列由SEQ ID No.15所示。
片断Rituximab单链抗体的核酸序列由SEQ ID No.17所示，VL和VH之间由连接肽2(SEQ ID No.43)连接。
片断Rituximab单链抗体-B7.1融合蛋白是由SEQ ID No.17所示的Rituximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ IDNo.1所示的B7.1核酸序列。
片断Rituximab单链抗体-B7.2融合蛋白是由SEQ ID No.17所示的Rituximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ IDNo.3所示的B7.2核酸序列。
片断Rituximab单链抗体-SEA融合蛋白是由SEQ ID No.17所示的Rituximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ IDNo.5所示的SEA核酸序列。
片断Rituximab单链抗体-B7.1-B7.2融合蛋白是由SEQ ID No.17所示的Rituximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.7所示的B7.1-B7.2融合蛋白核酸序列，Rituximab单链抗体-B7.1-B7.2融合蛋白的核酸序列由SEQ ID No.19所示。
片断Rituximab单链抗体-B7.1-B7.2-SEA融合蛋白是由SEQ ID No.17所示的Rituximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.9所示的B7.1-B7.2-SEA融合蛋白核酸序列，Rituximab单链抗体-B7.1-B7.2-SEA融合蛋白的核酸序列由SEQ ID No.21所示。
片断Cetuximab单链抗体的核酸序列由SEQ ID No.23所示，VL和VH之间由连接肽2(SEQ ID No.43)连接。
片断Cetuximab单链抗体-B7.1融合蛋白是由SEQ ID No.23所示的Cetuximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ IDNo.1所示的B7.1核酸序列。
片断Cetuximab单链抗体-B7.2融合蛋白是由SEQ ID No.23所示的Cetuximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ IDNo.3所示的B7.2核酸序列。
片断Cetuximab单链抗体-SEA融合蛋白是由SEQ ID No.23所示的Cetuximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ IDNo.5所示的SEA核酸序列。
片断Cetuximab单链抗体-B7.1-B7.2融合蛋白是由SEQ ID No.23所示的Cetuximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.7所示的B7.1-B7.2融合蛋白核酸序列，Cetuximab单链抗体-B7.1-B7.2融合蛋白的核酸序列由SEQ ID No.25所示。
片断Cetuximab单链抗体-B7.1-B7.2-SEA融合蛋白是由SEQ ID No.23所示的Cetuximab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.9所示的B7.1-B7.2-SEA融合蛋白核酸序列，Cetuximab单链抗体-B7.1-B7.2-SEA融合蛋白的核酸序列由SEQ ID No.27所示。
片断Panitumumab单链抗体的核酸序列由SEQ ID No.29所示，VL和VH之间由连接肽2(SEQ ID No.43)连接。
片断Panitumumab单链抗体-B7.1融合蛋白是由SEQ ID No.29所示的Panitumumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.1所示的B7.1核酸序列。
片断Panitumumab单链抗体-B7.2融合蛋白是由SEQ ID No.29所示的Panitumumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.3所示的B7.2核酸序列。
片断Panitumumab单链抗体-SEA融合蛋白是由SEQ ID No.29所示的Panitumumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.5所示的SEA核酸序列。
片断Panitumumab单链抗体-B7.1-B7.2融合蛋白是由SEQ ID No.29所示的Panitumumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.7所示的B7.1-B7.2融合蛋白核酸序列，Panitumumab单链抗体-B7.1-B7.2融合蛋白的核酸序列由SEQ ID No.31所示。
片断Panitumumab单链抗体-B7.1-B7.2-SEA融合蛋白是由SEQ ID No.29所示的Panitumumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.9所示的B7.1-B7.2-SEA融合蛋白核酸序列，Panitumumab单链抗体-B7.1-B7.2-SEA融合蛋白的核酸序列由SEQ ID No.33所示。
片断Ramucirumab单链抗体的核酸序列由SEQ ID No.35所示，VL和VH之间由连接肽2(SEQ ID No.43)连接。
片断Ramucirumab单链抗体-B7.1融合蛋白是由SEQ ID No.35所示的Ramucirumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.1所示的B7.1核酸序列。
片断Ramucirumab单链抗体-B7.2融合蛋白是由SEQ ID No.35所示的Ramucirumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.3所示的B7.2核酸序列。
片断Ramucirumab单链抗体-SEA融合蛋白是由SEQ ID No.35所示的Ramucirumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQID No.5所示的SEA核酸序列。
片断Ramucirumab单链抗体-B7.1-B7.2融合蛋白是由SEQ ID No.35所示的Ramucirumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.7所示的B7.1-B7.2融合蛋白核酸序列，Ramucirumab单链抗体-B7.1-B7.2融合蛋白的核酸序列由SEQ ID No.37所示。
片断Ramucirumab单链抗体-B7.1-B7.2-SEA融合蛋白是由SEQ ID No.35所示的Ramucirumab核酸序列后面连接一个由SEQ ID No.43所示的连接肽2核酸序列，这后面再连接一个由SEQ ID No.9所示的B7.1-B7.2-SEA融合蛋白核酸序列，Ramucirumab单链抗体-B7.1-B7.2-SEA融合蛋白的核酸序列由SEQ ID No.39所示。
在上述各个核酸序列片断的前面加入aattcggatccgaattcgagctcc碱基，这些增加的碱基小片断含有BamHI、EcoRI和SacI的限制酶切位点。
在上述各个核酸序列片断的后面加入aagcttgcggccgcactcgagcac碱基，这些增加的碱基小片断含有HindIII、NotI和XhoI的限制酶切位点。
利用所合成各个核酸序列的前面和后面的限制酶切位点把各种基因片断插入表达质粒pET22b(本发明以pET22b为例，但并不限定本发明，其它细菌、酵母或动物细胞等的表达质粒也可以用于本发明)，经DNA测序鉴定，这些重组的质粒及所表达的蛋白如下：
pET22b-Trastuzumab表达的是Trastuzumab单链抗体，氨基酸序列由SEQ IDNo.12所示；
pET22b-Trastuzumab-B7.1表达的是Trastuzumab单链抗体-B7.1融合蛋白；
pET22b-Trastuzumab-B7.2表达的是Trastuzumab单链抗体-B7.2融合蛋白；
pET22b-Trastuzumab-SEA表达的是Trastuzumab单链抗体-SEA融合蛋白；
pET22b-Trastuzumab-B7.1-B7.2表达的是Trastuzumab单链抗体-B7.1-B7.2融合蛋白，氨基酸序列由SEQ ID No.14所示；
pET22b-Trastuzumab-B7.1-B7.2-SEA表达的是Trastuzumab单链抗体-B7.1-B7.2-SEA融合蛋白，氨基酸序列由SEQ ID No.16所示；
pET22b-Rituximab表达的是Rituximab单链抗体，氨基酸序列由SEQ ID No.18所示；
pET22b-Rituximab-B7.1表达的是Rituximab单链抗体-B7.1融合蛋白；
pET22b-Rituximab-B7.2表达的是Rituximab单链抗体-B7.2融合蛋白；
pET22b-Rituximab-SEA表达的是Rituximab单链抗体-SEA融合蛋白；
pET22b-Rituximab-B7.1-B7.2表达的是Rituximab单链抗体-B7.1-B7.2融合蛋白，氨基酸序列由SEQ ID No.20所示；
pET22b-Rituximab-B7.1-B7.2-SEA表达的是Rituximab单链抗体-B7.1-B7.2-SEA融合蛋白，氨基酸序列由SEQ ID No.22所示；
pET22b-Cetuximab表达的是Cetuximab单链抗体，氨基酸序列由SEQ ID No.24所示；
pET22b-Cetuximab-B7.1表达的是Cetuximab单链抗体-B7.1融合蛋白；
pET22b-Cetuximab-B7.2表达的是Cetuximab单链抗体-B7.2融合蛋白；
pET22b-Cetuximab-SEA表达的是Cetuximab单链抗体-SEA融合蛋白；
pET22b-Cetuximab-B7.1-B7.2表达的是Cetuximab单链抗体-B7.1-B7.2融合蛋白，氨基酸序列由SEQ ID No.26所示；
pET22b-Cetuximab-B7.1-B7.2-SEA表达的是Cetuximab单链抗体-B7.1-B7.2-SEA融合蛋白，氨基酸序列由SEQ ID No.28所示；
pET22b-Panitumumab表达的是Panitumumab单链抗体，氨基酸序列由SEQ IDNo.30所示；
pET22b-Panitumumab-B7.1表达的是Panitumumab单链抗体-B7.1融合蛋白；
pET22b-Panitumumab-B7.2表达的是Panitumumab单链抗体-B7.2融合蛋白；
pET22b-Panitumumab-SEA表达的是Panitumumab单链抗体-SEA融合蛋白；
pET22b-Panitumumab-B7.1-B7.2表达的是Panitumumab单链抗体-B7.1-B7.2融合蛋白，氨基酸序列由SEQ ID No.32所示；
pET22b-Panitumumab-B7.1-B7.2-SEA表达的是Panitumumab单链抗体-B7.1-B7.2-SEA融合蛋白，氨基酸序列由SEQ ID No.34所示；
pET22b-Ramucirumab表达的是Ramucirumab单链抗体，氨基酸序列由SEQ IDNo.36所示；
pET22b-Ramucirumab-B7.1表达的是Ramucirumab单链抗体-B7.1融合蛋白；
pET22b-Ramucirumab-B7.2表达的是Ramucirumab单链抗体-B7.2融合蛋白；
pET22b-Ramucirumab-SEA表达的是Ramucirumab单链抗体-SEA融合蛋白；
pET22b-Ramucirumab-B7.1-B7.2表达的是Ramucirumab单链抗体-B7.1-B7.2融合蛋白，氨基酸序列由SEQ ID No.38所示；
pET22b-Ramucirumab-B7.1-B7.2-SEA表达的是Ramucirumab单链抗体-B7.1-B7.2-SEA融合蛋白，氨基酸序列由SEQ ID No.40所示。
实施例2各种蛋白的表达、变性和复性以及纯化
将各种表达质粒
pET22b-Trastuzumab、pET22b-Trastuzumab-B7.1、pET22b-Trastuzumab-B7.2、pET22b-Trastuzumab-SEA、pET22b-Trastuzumab-B7.1-B7.2、pET22b-Trastuzumab-B7.1-B7.2-SEA、pET22b-Rituximab、pET22b-Rituximab-B7.1、pET22b-Rituximab-B7.2、pET22b-Rituximab-SEA、pET22b-Rituximab-B7.1-B7.2、pET22b-Rituximab-B7.1-B7.2-SEA、pET22b-Cetuximab、pET22b-Cetuximab-B7.1、pET22b-Cetuximab-B7.2、pET22b-Cetuximab-SEA、pET22b-Cetuximab-B7.1-B7.2、pET22b-Cetuximab-B7.1-B7.2-SEA、pET22b-Panitumumab、pET22b-Panitumumab-B7.1、pET22b-Panitumumab-B7.2、pET22b-Panitumumab-SEA、pET22b-Panitumumab-B7.1-B7.2、pET22b-Panitumumab-B7.1-B7.2-SEA、pET22b-Ramucirumab、pET22b-Ramucirumab-B7.1、pET22b-Ramucirumab-B7.2、pET22b-Ramucirumab-SEA、pET22b-Ramucirumab-B7.1-B7.2和pET22b-Ramucirumab-B7.1-B7.2-SEA分别用电穿孔法转入大肠杆菌BL21(DE3)，利用抗生素Amp(Ampicillin)筛选阳性菌。接下来各种蛋白的表达、变性和复性以及纯化的过程大致相同，操作如下：
含有表达质粒的大肠杆菌BL21(DE3)先进行大规模37℃培养，然后加入IPTG(Isopropylthio-β-D-galactoside)使之浓度达到1mM并过夜30℃培养，从而诱导表达蛋白。第2天离心培养液和收集菌体，用超声波法破细胞壁，离心收集包含体沉淀，这里的蛋白是以包含体形式存在。包含体蛋白用6M尿素变性溶解，然后进行多阶段透析，透析溶液是逐步稀释的尿素例如3M、2M和1M，接下来是0.5M尿素，0.4M L-精氨酸，375μM氧化型谷胱甘肽GSSG，1.875mM还原型谷胱甘肽GSH，透析后进行离心，所得上清液就是蛋白的复性溶液。用His·Bind Purification Kit试剂盒(Novagen公司)对于蛋白进行纯化，先用BindingBuffer冲洗凝胶柱，同时也在蛋白溶液里加入Binding Buffer，将蛋白样品上柱，用WashBuffer漂洗，然后用Elute Buffer洗脱，用蛋白质电泳进行鉴定，将纯度为一条带的蛋白用于以后的实验。这样得到30种高纯度的Trastuzumab、Trastuzumab-B7.1、Trastuzumab-B7.2、Trastuzumab-SEA、Trastuzumab-B7.1-B7.2、Trastuzumab-B7.1-B7.2-SEA、Rituximab、Rituximab-B7.1、Rituximab-B7.2、Rituximab-SEA、Rituximab-B7.1-B7.2、Rituximab-B7.1-B7.2-SEA、Cetuximab、Cetuximab-B7.1、Cetuximab-B7.2、Cetuximab-SEA、Cetuximab-B7.1-B7.2、Cetuximab-B7.1-B7.2-SEA、Panitumumab、Panitumumab-B7.1、Panitumumab-B7.2、Panitumumab-SEA、Panitumumab-B7.1-B7.2、Panitumumab-B7.1-B7.2-SEA、Ramucirumab、Ramucirumab-B7.1、Ramucirumab-B7.2、Ramucirumab-SEA、Ramucirumab-B7.1-B7.2和Ramucirumab-B7.1-B7.2-SEA蛋白。
实施例3制备表达人源HER2(ErbB-2)的小鼠肉瘤细胞S180细胞株
根据GenBank数据库(NM_004448.3)将人HER2(ErbB-2)基因片断插入pEGFP-N1质粒(Clontech公司)，将pEGFP-N1-HER2(erbB-2)的质粒DNA(ug)与转染试剂Lipofectamine2000(Invitrogen公司)(ul)以1:2的比率混合，然后将混合物加入到含有小鼠肉瘤细胞S180的24孔板中培养(1x105个细胞/孔)，第2天用含G418抗生素(浓度为500ug/ml)的培养基继续培养和筛选(有限稀释法)，2周后在荧光显微镜下挑选抗性单克隆细胞株，并用小鼠抗人HER2(ErbB-2)抗体(Abcam公司)确认单克隆细胞株表达人源HER2(ErbB-2)，这样就得到了S180/HER2(ErbB-2)单克隆细胞株。
实施例4制备表达人源CD20的小鼠肉瘤细胞S180细胞株
根据参考论文(Proc Natl Acad Sci USA,85,208-212,1988；J Exp Med,167,1975-1980,1988)和GenBank数据库(M27394.1、X12530.1)将人CD20基因片断插入pEGFP-N1质粒(Clontech公司)，将pEGFP-N1-CD20的质粒DNA(ug)与转染试剂Lipofectamine2000(Invitrogen公司)(ul)以1:2的比率混合，然后将混合物加入到含有小鼠肉瘤细胞S180的24孔板中培养(1x105个细胞/孔)，第2天用含G418抗生素(浓度为500ug/ml)的培养基继续培养和筛选(有限稀释法)，2周后在荧光显微镜下挑选抗性单克隆细胞株，并用小鼠抗人CD20抗体(Abcam公司)确认单克隆细胞株表达人源CD20，这样就得到了S180/CD20单克隆细胞株。
实施例5用正常小鼠开展的抑制肿瘤实验
实验一
选择雄性ICR小鼠，4-5周，18-22g，分成7组，每组30只小鼠。表达HER2(ErbB-2)的S180/HER2(ErbB-2)肿瘤细胞来自于小鼠腹水培养，将3x106数量的S180/HER2(ErbB-2)肿瘤细胞接种于小鼠的右侧腋下，然后每隔2天(第2天、第5天、第8天)往尾静脉内分别注射Trastuzumab、Trastuzumab-B7.1、Trastuzumab-B7.2、Trastuzumab-SEA、Trastuzumab-B7.1-B7.2和Trastuzumab-B7.1-B7.2-SEA蛋白溶液0.2ml(100pmol)/只，而对照组注射等量生理盐水(0.9％NaCl)，第11天处死小鼠，观察小鼠肿瘤生长情况以及测量处死小鼠后肿瘤的重量。
图1表示各种蛋白抑制S180/HER2(ErbB-2)肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中右边曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Trastuzumab组，3是Trastuzumab-B7.1组，4是Trastuzumab-B7.2组，5是Trastuzumab-SEA组，6是Trastuzumab-B7.1-B7.2组、7是Trastuzumab-B7.1-B7.2-SEA组。
图2表示第11天杀死小鼠解剖后的S180/HER2(ErbB-2)肿瘤重量，横轴表示实验用小鼠的编组，1是生理盐水组，2是Trastuzumab组，3是Trastuzumab-B7.1组，4是Trastuzumab-B7.2组，5是Trastuzumab-SEA组，6是Trastuzumab-B7.1-B7.2组、7是Trastuzumab-B7.1-B7.2-SEA组。
结果表明各种蛋白都可以有效抑制肿瘤生长，而Trastuzumab-B7.1-B7.2抑制肿瘤的效果更显著，Trastuzumab-B7.1-B7.2-SEA抑制肿瘤的效果最好。
实验二
选择雄性ICR小鼠，4-5周，18-22g，分成7组，每组30只小鼠。表达CD20的S180/CD20肿瘤细胞来自于小鼠腹水培养，将3x106数量的S180/CD20肿瘤细胞接种于小鼠的右侧腋下，然后每隔2天(第2天、第5天、第8天)往尾静脉内分别注射Rituximab、Rituximab-B7.1、Rituximab-B7.2、Rituximab-SEA、Rituximab-B7.1-B7.2和Rituximab-B7.1-B7.2-SEA蛋白溶液0.2ml(100pmol)/只，而对照组注射等量生理盐水(0.9％NaCl)，第11天处死小鼠，观察小鼠肿瘤生长情况以及测量处死小鼠后肿瘤的重量。
图3表示各种蛋白抑制S180/CD20肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中右边曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Rituximab组，3是Rituximab-B7.1组，4是Rituximab-B7.2组，5是Rituximab-SEA组，6是Rituximab-B7.1-B7.2组，7是Rituximab-B7.1-B7.2-SEA组。
图4表示第11天杀死小鼠解剖后的S180/CD20肿瘤重量，横轴表示实验用小鼠的编组，1是生理盐水组，2是Rituximab组，3是Rituximab-B7.1组，4是Rituximab-B7.2组，5是Rituximab-SEA组，6是Rituximab-B7.1-B7.2组，7是Rituximab-B7.1-B7.2-SEA组。
结果表明各种蛋白都可以有效抑制肿瘤生长，而Rituximab-B7.1-B7.2抑制肿瘤的效果更显著，Rituximab-B7.1-B7.2-SEA抑制肿瘤的效果最好。
实施例6肿瘤组织内T淋巴细胞的检测
在上面实施例5的实验一的中挑选生理盐水组、Trastuzumab组、Trastuzumab-B7.1组、Trastuzumab-B7.2组、Trastuzumab-SEA组、Trastuzumab-B7.1-B7.2和Trastuzumab-B7.1-B7.2-SEA组的携带S180/HER2(ErbB-2)肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内T淋巴细胞的样品。
同样在在上面实施例5的实验二中挑选生理盐水组、Rituximab组、Rituximab-B7.1组、Rituximab-B7.2组、Rituximab-SEA组、Rituximab-B7.1-B7.2组和Rituximab-B7.1-B7.2-SEA组的携带S180/CD20肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内T淋巴细胞的样品。
肿瘤组织切成小块，用石蜡包埋制成石蜡切片，然后进行免疫组织化学实验。为了检测肿瘤组织内的T细胞，抗T细胞的特异性抗体采用Santa Cruz Biotechnolog公司的兔抗小鼠抗CD3抗体，然后用二抗以及avidin-biotin-perxidase complex(Zymed公司)，最后用Diaminobenzidine(DAB)显色，以后下面的实验中的二抗和DAB显色采用类似的方法。
生理盐水组和Trastuzumab组的肿瘤内几乎没有T细胞，所以图5只提供Trastuzumab组的数据。Trastuzumab-B7.1组、Trastuzumab-B7.2组和Trastuzumab-SEA组的肿瘤内的T细胞数量差不多，所以图5只提供Trastuzumab-B7.1组的数据。
图5利用常规免疫组化检测携带S180/HER2(ErbB-2)肿瘤组织内的T细胞，棕色小点部分就是T细胞，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
结果显示Trastuzumab-B7.1-B7.2组的肿瘤组织内有许多T细胞，Trastuzumab-B7.1-B7.2-SEA组的肿瘤组织内的T细胞最多(有大量棕色小点)。
生理盐水组和Rituximab组的肿瘤内几乎没有T细胞，所以图6只提供Rituximab组的数据。Rituximab-B7.1组、Rituximab-B7.2组和Rituximab-SEA组的肿瘤内的T细胞数量差不多，所以图6只提供Rituximab-B7.1组的数据。
图6利用常规免疫组化检测携带S180/CD20肿瘤组织内的T细胞，棕色小点部分就是T细胞，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
结果显示Rituximab-B7.1-B7.2组的肿瘤组织内有许多T细胞，Rituximab-B7.1-B7.2-SEA组的肿瘤组织内的T细胞最多(有大量棕色小点)。
实施例7肿瘤组织内干扰素-γ(IFN-γ)的检测
在上面实施例5的实验一的中挑选生理盐水组、Trastuzumab组、Trastuzumab-B7.1组、Trastuzumab-B7.2组、Trastuzumab-SEA组、Trastuzumab-B7.1-B7.2和Trastuzumab-B7.1-B7.2-SEA组的携带S180/HER2(ErbB-2)肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内干扰素-γ(IFN-γ)的样品。
同样在在上面实施例5的实验二中挑选生理盐水组、Rituximab组、Rituximab-B7.1组、Rituximab-B7.2组、Rituximab-SEA组、Rituximab-B7.1-B7.2组和Rituximab-B7.1-B7.2-SEA组的携带S180/CD20肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内干扰素-γ(IFN-γ)的样品。
用免疫组织化学方法检测肿瘤组织内由T细胞分泌的细胞因子IFN-γ，抗体是Santa Cruz Biotechnolog公司的兔抗小鼠抗IFN-γ抗体。
生理盐水组和Trastuzumab组的肿瘤内几乎没有IFN-γ，所以图7只提供Trastuzumab组的数据。Trastuzumab-B7.1组、Trastuzumab-B7.2组和Trastuzumab-SEA组的肿瘤内的IFN-γ差不多，所以图7只提供Trastuzumab-B7.1组的数据。
图7利用常规免疫组化检测携带S180/HER2(ErbB-2)肿瘤组织内的干扰素-γ(IFN-γ)，棕色部分就是T细胞分泌的IFN-γ，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
结果显示Trastuzumab-B7.1-B7.2组的肿瘤组织内有许多IFN-γ，Trastuzumab-B7.1-B7.2-SEA组的肿瘤组织内有更多IFN-γ。
生理盐水组和Rituximab组的肿瘤内几乎没有IFN-γ，所以图8只提供Rituximab组的数据。Rituximab-B7.1组、Rituximab-B7.2组和Rituximab-SEA组的肿瘤内的IFN-γ差不多，所以图8只提供Rituximab-B7.1组的数据。
图8利用常规免疫组化检测携带S180/CD20肿瘤组织内的干扰素-γ(IFN-γ)，棕色部分就是T细胞分泌的IFN-γ，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
结果显示Rituximab-B7.1-B7.2组的肿瘤组织内有许多IFN-γ，Rituximab-B7.1-B7.2-SEA组的肿瘤组织内有更多IFN-γ。
实施例8肿瘤组织内颗粒酶(Granzyme B)的检测
在上面实施例5的实验一的中挑选生理盐水组、Trastuzumab组、Trastuzumab-B7.1组、Trastuzumab-B7.2组、Trastuzumab-SEA组、Trastuzumab-B7.1-B7.2和Trastuzumab-B7.1-B7.2-SEA组的携带S180/HER2(ErbB-2)肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内颗粒酶(Granzyme B)的样品。
同样在在上面实施例5的实验二中挑选生理盐水组、Rituximab组、Rituximab-B7.1组、Rituximab-B7.2组、Rituximab-SEA组、Rituximab-B7.1-B7.2组和Rituximab-B7.1-B7.2-SEA组的携带S180/CD20肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内颗粒酶(Granzyme B)的样品。
用免疫组织化学方法检测肿瘤组织内由T细胞分泌的颗粒酶，抗体是Abcam公司的兔抗小鼠抗Granzyme B抗体。
生理盐水组和Trastuzumab组的肿瘤内几乎没有颗粒酶，所以图9只提供Trastuzumab组的数据。Trastuzumab-B7.1组、Trastuzumab-B7.2组和Trastuzumab-SEA组的肿瘤内的颗粒酶差不多，所以图9只提供Trastuzumab-B7.1组的数据。
图9利用常规免疫组化检测颗粒酶(Granzyme B)，棕色部分就是T细胞分泌的颗粒酶，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
结果显示Trastuzumab-B7.1-B7.2组的肿瘤组织内有许多颗粒酶，Trastuzumab-B7.1-B7.2-SEA组的肿瘤组织内有更多颗粒酶。
生理盐水组和Rituximab组的肿瘤内几乎没有颗粒酶，所以图10只提供Rituximab组的数据。Rituximab-B7.1组、Rituximab-B7.2组和Rituximab-SEA组的肿瘤内的颗粒酶差不多，所以图10只提供Rituximab-B7.1组的数据。
图10利用常规免疫组化检测颗粒酶(Granzyme B)，棕色部分就是T细胞分泌的颗粒酶，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
结果显示Rituximab-B7.1-B7.2组的肿瘤组织内有许多颗粒酶，Rituximab-B7.1-B7.2-SEA组的肿瘤组织内有更多颗粒酶。
实施例9常规免疫组化检测肿瘤细胞染色体的断裂(TUNEL分析)
在上面实施例5的实验一的中挑选生理盐水组、Trastuzumab组、Trastuzumab-B7.1组、Trastuzumab-B7.2组、Trastuzumab-SEA组、Trastuzumab-B7.1-B7.2和Trastuzumab-B7.1-B7.2-SEA组的携带S180/HER2(ErbB-2)肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内肿瘤细胞染色体断裂(TUNEL分析)的样品。
同样在在上面实施例5的实验二中挑选生理盐水组、Rituximab组、Rituximab-B7.1组、Rituximab-B7.2组、Rituximab-SEA组、Rituximab-B7.1-B7.2组和Rituximab-B7.1-B7.2-SEA组的携带S180/CD20肿瘤小鼠解剖后的肿瘤组织作为这里检测肿瘤组织内肿瘤细胞染色体断裂(TUNEL分析)的样品。
用免疫组织化学方法检测肿瘤组织内肿瘤细胞的染色体是否断裂，从而判断肿瘤细胞是否死亡，TUNEL分析试剂盒来自罗氏公司。
生理盐水组和Trastuzumab组的肿瘤内几乎没有死亡的肿瘤细胞，所以图11只提供Trastuzumab组的数据。Trastuzumab-B7.1组、Trastuzumab-B7.2组和Trastuzumab-SEA组的肿瘤内死亡的肿瘤细胞差不多，所以图11只提供Trastuzumab-B7.1组的数据。
图11利用常规免疫组化检测染色体断裂，棕色部分就是发生染色体断裂的肿瘤细胞，1是Trastuzumab组，2是Trastuzumab-B7.1组，3是Trastuzumab-B7.1-B7.2组，4是Trastuzumab-B7.1-B7.2-SEA组。
结果显示Trastuzumab-B7.1-B7.2组的肿瘤内有许多肿瘤细胞死亡，Trastuzumab-B7.1-B7.2-SEA组的肿瘤内有更多肿瘤细胞死亡。
生理盐水组和Rituximab组的肿瘤内几乎没有死亡的肿瘤细胞，所以图12只提供Rituximab组的数据。Rituximab-B7.1组、Rituximab-B7.2组和Rituximab-SEA组的肿瘤内死亡的肿瘤细胞差不多，所以图12只提供Rituximab-B7.1组的数据。
图12利用常规免疫组化检测染色体断裂，棕色部分就是发生染色体断裂的肿瘤细胞，1是Rituximab组，2是Rituximab-B7.1组，3是Rituximab-B7.1-B7.2组，4是Rituximab-B7.1-B7.2-SEA组。
结果显示Rituximab-B7.1-B7.2组的肿瘤内有许多肿瘤细胞死亡，Rituximab-B7.1-B7.2-SEA组的肿瘤内有更多肿瘤细胞死亡。
实施例10抑制人肿瘤生长
实验材料准备
人肿瘤细胞：人表皮癌细胞株A431和人肺癌细胞株HCC4006。
小鼠：免疫缺陷型CB17小鼠(SCID鼠)
人T淋巴细胞：人外周血细胞购自天津血液中心，用Ficoll密度离心和尼龙毛柱方法取得T淋巴细胞(J Clin Invest,91,1490-1498,1993)。
实验一
选择雌性SCID鼠，6周，18-22g，分成7组，每组30只小鼠。A431癌细胞来自于SCID鼠腹水培养，将1x106A431癌细胞接种于SCID鼠的右侧腋下，第1、天往尾静脉内注射2x108人T细胞，然后第2天起隔天往尾静脉内注射Cetuximab、Cetuximab-B7.1、Cetuximab-B7.2、Cetuximab-SEA、Cetuximab-B7.1-B7.2和Cetuximab-B7.1-B7.2-SEA蛋白溶液0.2ml(150pmol)/只，而对照组注射等量生理盐水(0.9％NaCl)，第9天处死小鼠，观察小鼠肿瘤生长情况以及测量处死小鼠后肿瘤的重量。
图13表示抗体抑制肿瘤生长和第9天杀死荷瘤小鼠解剖后的肿瘤重量。
A表示各种蛋白抑制A431肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中上面曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Cetuximab组，3是Cetuximab-B7.1组，4是Cetuximab-B7.2组，5是Cetuximab-SEA组，6是Cetuximab-B7.1-B7.2组，7是Cetuximab-B7.1-B7.2-SEA组。
B表示第9天杀死小鼠解剖后的A431肿瘤重量，横轴表示的是实验用小鼠的编组，1是生理盐水组，2是Cetuximab组，3是Cetuximab-B7.1组，4是Cetuximab-B7.2组，5是Cetuximab-SEA组，6是Cetuximab-B7.1-B7.2组，7是Cetuximab-B7.1-B7.2-SEA组。
结果表明各种蛋白都可以有效抑制肿瘤生长，而Cetuximab-B7.1-B7.2抑制肿瘤的效果更显著，Cetuximab-B7.1-B7.2-SEA抑制肿瘤的效果最好。
实验二
选择雌性SCID鼠，6周，18-22g，分成7组，每组30只小鼠。A431癌细胞来自于SCID鼠腹水培养，将1x106A431癌细胞接种于SCID鼠的右侧腋下，第1天往尾静脉内注射2x108人T细胞，然后第2天起隔天往尾静脉内注射Panitumumab、Panitumumab-B7.1、Panitumumab-B7.2、Panitumumab-SEA、Panitumumab-B7.1-B7.2和Panitumumab-B7.1-B7.2-SEA蛋白溶液0.2ml(150pmol)/只，而对照组注射等量生理盐水(0.9％NaCl)，第9天处死小鼠，观察小鼠肿瘤生长情况以及测量处死小鼠后肿瘤的重量。
图14表示抗体抑制肿瘤生长和第9天杀死荷瘤小鼠解剖后的肿瘤重量。
A表示各种蛋白抑制A431肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中上面曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Panitumumab组，3是Panitumumab-B7.1组，4是Panitumumab-B7.2组，5是Panitumumab-SEA组，6是Panitumumab-B7.1-B7.2组，7是Panitumumab-B7.1-B7.2-SEA组。
B表示第9天杀死小鼠解剖后的A431肿瘤重量，横轴表示的是实验用小鼠的编组，1是生理盐水组，2是Panitumumab组，3是Panitumumab-B7.1组，4是Panitumumab-B7.2组，5是Panitumumab-SEA组，6是Panitumumab-B7.1-B7.2组，7是Panitumumab-B7.1-B7.2-SEA组。
结果表明各种蛋白都可以有效抑制肿瘤生长，而Panitumumab-B7.1-B7.2抑制肿瘤的效果更显著，Panitumumab-B7.1-B7.2-SEA抑制肿瘤的效果最好。
实验三
选择雌性SCID鼠，6周，18-22g，分成7组，每组30只小鼠。HCC4006癌细胞来自于SCID鼠腹水培养，将1x106HCC4006癌细胞接种于SCID鼠的右侧腋下，第1天往尾静脉内注射2x108人T细胞，然后第2天起隔天往尾静脉内注射Ramucirumab、Ramucirumab-B7.1、Ramucirumab-B7.2、Ramucirumab-SEA、Ramucirumab-B7.1-B7.2和Ramucirumab-B7.1-B7.2-SEA蛋白溶液0.2ml(150pmol)/只，而对照组注射等量生理盐水(0.9％NaCl)，第9天处死小鼠，观察小鼠肿瘤生长情况以及测量处死小鼠后肿瘤的重量。
图15表示抗体抑制肿瘤生长和第9天杀死荷瘤小鼠解剖后的肿瘤重量。
A表示各种蛋白抑制HCC4006肿瘤生长，图中横轴表示荷瘤小鼠生长天数，纵轴表示肿瘤体积(cm3)，图中上面曲线编号表示实验用小鼠的编组，1是生理盐水组，2是Ramucirumab组，3是Ramucirumab-B7.1组，4是Ramucirumab-B7.2组，5是Ramucirumab-SEA组，6是Ramucirumab-B7.1-B7.2，7是Ramucirumab-B7.1-B7.2-SEA组。
B表示第9天杀死小鼠解剖后的HCC4006肿瘤重量，横轴表示的是实验用小鼠的编组，1是生理盐水组，2是Ramucirumab组，3是Ramucirumab-B7.1组，4是Ramucirumab-B7.2组，5是Ramucirumab-SEA组，6是Ramucirumab-B7.1-B7.2，7是Ramucirumab-B7.1-B7.2-SEA组。
结果表明各种蛋白都可以有效抑制肿瘤生长，而Ramucirumab-B7.1-B7.2抑制肿瘤的效果更显著，Ramucirumab-B7.1-B7.2-SEA抑制肿瘤的效果最好。
T细胞可以通过多种手段来杀死肿瘤，例如分泌凋亡因子TNF-α和干扰素-γ(Interferon-γ，IFN-γ)、诱导肿瘤细胞表达凋亡因子Fas来导致肿瘤细胞的死亡、分泌穿孔素和颗粒酶进入肿瘤细胞来杀死肿瘤细胞(cell Mol Immunol,6,15-25,2009；ImmunolToday,16,194-201,1995；Cell,46,641-642,1986；Immunol Today,16,202-206,1995；AnnHematol,84,627-639,2005；Apoptosis,8,237-249,2003)，B7.1、B7.2和SEA分子都可以诱导并激活T细胞，构建多种融合蛋白含有B7.1、B7.2或SEA都可以强化诱导并激活T细胞的能力，而B7.1-B7.2-SEA融合蛋白则发挥最大的这样能力。
抗体本身不利用T细胞，一种抗肿瘤抗体如果连接B7.1、B7.2、SEA、B7.1-B7.2或B7.1-B7.2-SEA，则赋予抗体拥有肿瘤靶向诱导并激活T细胞，使得这些T细胞杀死肿瘤，从而大大增强了抗体药物在肿瘤治疗的效果。
肿瘤细胞通过其自身上的PD-L1分子与T细胞上的PD-1分子相互作用而导致T细胞凋亡，这样肿瘤就逃避了免疫监视以及T细胞对于肿瘤的攻击(Drug Design DevelopTher,9,2883-2886,2015；Cancer Treat Rev,54,58-67,2017；Int J Infect Dis,56,221-228,2017；J Biomed Sci,24,26,2017)，为了解决肿瘤细胞躲避免疫监视而引起T细胞凋亡，本发明通过一种癌靶向增强型抗肿瘤融合蛋白诱导并激活T细胞，增加T细胞的数量，增强T细胞的活性，使得这些大量有活性的T细胞攻击肿瘤。这项技术不局限于某一特定的肿瘤，通过B7.1-B7.2或B7.1-B7.2-SEA融合蛋白与各种抗体连接而形成新融合蛋白就可以广泛应用于各种实体肿瘤和血液肿瘤等癌症的治疗。
一种癌靶向增强型抗肿瘤融合蛋白衍生物，其特征在于根据B7.1-B7.2或B7.1-B7.2-SEA融合蛋白与各种抗体连接而形成新融合蛋白中的B7.1-B7.2或B7.1-B7.2-SEA融合蛋白所产生的点突变、氨基酸序列缺失或导入突变体，这样可以更大幅度提高新融合蛋白衍生物诱导并激活T细胞的能力，从而更有效地杀死肿瘤。一种癌靶向增强型抗肿瘤融合蛋白，其连接方式不限于目前连接方式。
作为药物其剂型可以是乳化剂、脂质体、分散剂、稳定剂等一起制成各种注射、口服、敷贴以及手术处理等药物的给药形式。除了融合蛋白质本身可以作为药物外，编码融合蛋白质的核苷酸片段或载体还可以作为基因治疗形式来应用。
以上实施例和优选实施方式的描述属于对权利要求限定的本发明的示意性说明，而不是限制本发明。需要特别指出的是，只要不脱离本发明的宗旨，所有显而易见的改变以及具有等价代换的相似发明，均包含在本发明的保护范围之内。
序列表
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Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu His
85 90 95
Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro Ser
100 105 110
Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile Cys
115 120 125
Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu Asn
130 135 140
Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro Glu
145 150 155 160
Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr Thr
165 170 175
Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val Asn
180 185 190
Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp Asn
195 200 205
Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln Ala
210 215 220
Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser Gln
225 230 235 240
Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu Asn
245 250 255
Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser Val
260 265 270
His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp Thr
275 280 285
Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln Cys
290 295 300
Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln Met
305 310 315 320
Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile Val
325 330 335
Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys Ser
340 345 350
Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu Arg
355 360 365
Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser Gln
370 375 380
Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val Ser
385 390 395 400
Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu Thr
405 410 415
Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu Asp
420 425 430
Pro Gln Pro Pro Pro Asp His
435
<210> 9
<211> 2043
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
gttatccacg tgaccaagga agtgaaagaa gtggcaacgc tgtcctgtgg tcacaatgtt 60
tctgttgaag agctggcaca aactcgcatc tactggcaaa aggagaagaa aatggtgctg 120
actatgatgt ctggggacat gaatatatgg cccgagtaca agaaccggac catctttgat 180
atcactaata acctctccat tgtgatcctg gctctgcgcc catctgacga gggcacatac 240
gagtgtgttg ttctgaagta tgaaaaagac gctttcaagc gggaacacct ggctgaagtg 300
acgttatcag tcaaagctga cttccctaca cctagtatat ctgactttga aattccaact 360
tctaatatta gaaggataat ttgctcaacc tctggaggtt ttccagagcc tcacctctcc 420
tggttggaaa atggagaaga attaaatgcc atcaacacaa cagtttccca agatcctgaa 480
actgagctct atgctgttag cagcaaactg gatttcaata tgacaaccaa ccacagcttc 540
atgtgtctca tcaagtatgg acatttaaga gtgaatcaga ccttcaactg gaatacaacc 600
aagcaagagc attttcctga taacgtcgac aagctttccg gcggaggtgg cgctcctctg 660
aagattcaag cttatttcaa tgagactgca gacctgccat gccaatttgc aaactctcaa 720
aaccaaagcc tgagtgagct agtagtattt tggcaggacc aggaaaactt ggttctgaat 780
gaggtatact taggcaaaga gaaatttgac agtgttcatt ccaagtatat gggccgcaca 840
agttttgatt cggacagttg gaccctgaga cttcacaatc ttcagatcaa ggacaagggc 900
ttgtatcaat gtatcatcca tcacaaaaag cccacaggaa tgattcgcat ccaccagatg 960
aattctgaac tgtcagtgct tgctaacttc agtcaacctg aaatagtacc aatttctaat 1020
ataacagaaa atgtgtacat aaatttgacc tgctcatcta tacacggtta cccagaacct 1080
aagaagatga gtgttttgct aagaaccaag aattcaacta tcgagtatga tggtattatg 1140
cagaaatctc aagataatgt cacagaactg tacgacgttt ccatcagctt gtctgtttca 1200
ttccctgatg ttacgagcaa tatgaccatc ttctgtattc tggaaactga caagacgcgg 1260
cttttatctt cacctttctc tatagagctt gaggaccctc agcctccccc agaccacgtc 1320
gacaagcttt ccggcggagg tggcagcgag aaaagcgaag aaataaatga aaaagatttg 1380
cgaaaaaagt ctgaattgca gggaacagct ttaggcaatc ttaaacaaat ctattattac 1440
aatgaaaaag ctaaaactga aaataaagag agtcacgatc aatttttaca gcatactata 1500
ttgtttaaag gcttttttac agatcattcg tggtataacg atttattagt agattttgat 1560
tcaaaggata ttgttgataa atataaaggg aaaaaagtag acttgtatgg tgcttattat 1620
ggttatcaat gtgcgggtgg tacaccaaac aaaacagctt gtatgtatgg tggtgtaacg 1680
ttacatgata ataatcgatt gaccgaagag aaaaaagtgc cgatcaattt atggctagac 1740
ggtaaacaaa atacagtacc tttggaaacg gttaaaacga ataagaaaaa tgtaactgtt 1800
caggagttgg atcttcaagc aagacgttat ttacaggaaa aatataattt atataactct 1860
gatgtttttg atgggaaggt tcagagggga ttaatcgtgt ttcatacttc tacagaacct 1920
tcggttaatt acgatttatt tggtgctcaa ggacagtatt caaatacact attaagaata 1980
tatagagata ataaaacgat taactctgaa aacatgcata ttgatatata tttatataca 2040
agt 2043
<210> 10
<211> 681
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser Cys
1 5 10 15
Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr Trp
20 25 30
Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met Asn
35 40 45
Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn Asn
50 55 60
Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr Tyr
65 70 75 80
Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu His
85 90 95
Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro Ser
100 105 110
Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile Cys
115 120 125
Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu Asn
130 135 140
Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro Glu
145 150 155 160
Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr Thr
165 170 175
Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val Asn
180 185 190
Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp Asn
195 200 205
Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln Ala
210 215 220
Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser Gln
225 230 235 240
Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu Asn
245 250 255
Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser Val
260 265 270
His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp Thr
275 280 285
Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln Cys
290 295 300
Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln Met
305 310 315 320
Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile Val
325 330 335
Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys Ser
340 345 350
Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu Arg
355 360 365
Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser Gln
370 375 380
Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val Ser
385 390 395 400
Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu Thr
405 410 415
Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu Asp
420 425 430
Pro Gln Pro Pro Pro Asp His Val Asp Lys Leu Ser Gly Gly Gly Gly
435 440 445
Ser Glu Lys Ser Glu Glu Ile Asn Glu Lys Asp Leu Arg Lys Lys Ser
450 455 460
Glu Leu Gln Gly Thr Ala Leu Gly Asn Leu Lys Gln Ile Tyr Tyr Tyr
465 470 475 480
Asn Glu Lys Ala Lys Thr Glu Asn Lys Glu Ser His Asp Gln Phe Leu
485 490 495
Gln His Thr Ile Leu Phe Lys Gly Phe Phe Thr Asp His Ser Trp Tyr
500 505 510
Asn Asp Leu Leu Val Asp Phe Asp Ser Lys Asp Ile Val Asp Lys Tyr
515 520 525
Lys Gly Lys Lys Val Asp Leu Tyr Gly Ala Tyr Tyr Gly Tyr Gln Cys
530 535 540
Ala Gly Gly Thr Pro Asn Lys Thr Ala Cys Met Tyr Gly Gly Val Thr
545 550 555 560
Leu His Asp Asn Asn Arg Leu Thr Glu Glu Lys Lys Val Pro Ile Asn
565 570 575
Leu Trp Leu Asp Gly Lys Gln Asn Thr Val Pro Leu Glu Thr Val Lys
580 585 590
Thr Asn Lys Lys Asn Val Thr Val Gln Glu Leu Asp Leu Gln Ala Arg
595 600 605
Arg Tyr Leu Gln Glu Lys Tyr Asn Leu Tyr Asn Ser Asp Val Phe Asp
610 615 620
Gly Lys Val Gln Arg Gly Leu Ile Val Phe His Thr Ser Thr Glu Pro
625 630 635 640
Ser Val Asn Tyr Asp Leu Phe Gly Ala Gln Gly Gln Tyr Ser Asn Thr
645 650 655
Leu Leu Arg Ile Tyr Arg Asp Asn Lys Thr Ile Asn Ser Glu Asn Met
660 665 670
His Ile Asp Ile Tyr Leu Tyr Thr Ser
675 680
<210> 11
<211> 726
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
gatatccaga tgacccagtc cccgagctcc ctgtccgcct ctgtgggcga tagggtcact 60
atcacctgcc gtgccagtca ggatgtgaat actgctgtag cctggtatca acagaaaccc 120
ggaaaggccc cgaaactgct gatttactcg gcatccttcc tctactctgg agtcccttct 180
cgcttctctg gttcccgctc tgggacggat ttcactctga ccatcagctc cctgcagccg 240
gaagacttcg caacttatta ctgtcagcaa cactatacta ctcctccgac gttcggacag 300
ggtaccaagg tggagatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctgagg ttcagctggt ggagtctggc ggtggcctgg tgcagcccgg gggctctctc 420
cgtttgtcct gtgcagcttc tggcttcaac attaaagaca cctatatcca ctgggtgcgt 480
caggctccgg gtaagggcct ggagtgggtt gcaaggattt atcctacgaa tggttatact 540
cgttatgccg atagcgtcaa gggccgtttc actataagcg cagacacttc gaaaaacaca 600
gcctacctcc agatgaacag cctgcgtgct gaggacactg ccgtctatta ttgtagcaga 660
tggggtgggg acggcttcta tgctatggac tactggggtc aaggtacact agtcaccgtc 720
agcagc 726
<210> 12
<211> 242
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu
115 120 125
Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His Trp Val Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Tyr Pro Thr
165 170 175
Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile
180 185 190
Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp Gly Gly Asp
210 215 220
Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
225 230 235 240
Ser Ser
<210> 13
<211> 2088
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
gatatccaga tgacccagtc cccgagctcc ctgtccgcct ctgtgggcga tagggtcact 60
atcacctgcc gtgccagtca ggatgtgaat actgctgtag cctggtatca acagaaaccc 120
ggaaaggccc cgaaactgct gatttactcg gcatccttcc tctactctgg agtcccttct 180
cgcttctctg gttcccgctc tgggacggat ttcactctga ccatcagctc cctgcagccg 240
gaagacttcg caacttatta ctgtcagcaa cactatacta ctcctccgac gttcggacag 300
ggtaccaagg tggagatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctgagg ttcagctggt ggagtctggc ggtggcctgg tgcagcccgg gggctctctc 420
cgtttgtcct gtgcagcttc tggcttcaac attaaagaca cctatatcca ctgggtgcgt 480
caggctccgg gtaagggcct ggagtgggtt gcaaggattt atcctacgaa tggttatact 540
cgttatgccg atagcgtcaa gggccgtttc actataagcg cagacacttc gaaaaacaca 600
gcctacctcc agatgaacag cctgcgtgct gaggacactg ccgtctatta ttgtagcaga 660
tggggtgggg acggcttcta tgctatggac tactggggtc aaggtacact agtcaccgtc 720
agcagcggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccac 2088
<210> 14
<211> 696
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu
115 120 125
Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His Trp Val Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Tyr Pro Thr
165 170 175
Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile
180 185 190
Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp Gly Gly Asp
210 215 220
Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
225 230 235 240
Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His
690 695
<210> 15
<211> 2814
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
gatatccaga tgacccagtc cccgagctcc ctgtccgcct ctgtgggcga tagggtcact 60
atcacctgcc gtgccagtca ggatgtgaat actgctgtag cctggtatca acagaaaccc 120
ggaaaggccc cgaaactgct gatttactcg gcatccttcc tctactctgg agtcccttct 180
cgcttctctg gttcccgctc tgggacggat ttcactctga ccatcagctc cctgcagccg 240
gaagacttcg caacttatta ctgtcagcaa cactatacta ctcctccgac gttcggacag 300
ggtaccaagg tggagatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctgagg ttcagctggt ggagtctggc ggtggcctgg tgcagcccgg gggctctctc 420
cgtttgtcct gtgcagcttc tggcttcaac attaaagaca cctatatcca ctgggtgcgt 480
caggctccgg gtaagggcct ggagtgggtt gcaaggattt atcctacgaa tggttatact 540
cgttatgccg atagcgtcaa gggccgtttc actataagcg cagacacttc gaaaaacaca 600
gcctacctcc agatgaacag cctgcgtgct gaggacactg ccgtctatta ttgtagcaga 660
tggggtgggg acggcttcta tgctatggac tactggggtc aaggtacact agtcaccgtc 720
agcagcggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccacgt cgacaagctt 2100
tccggcggag gtggcagcga gaaaagcgaa gaaataaatg aaaaagattt gcgaaaaaag 2160
tctgaattgc agggaacagc tttaggcaat cttaaacaaa tctattatta caatgaaaaa 2220
gctaaaactg aaaataaaga gagtcacgat caatttttac agcatactat attgtttaaa 2280
ggctttttta cagatcattc gtggtataac gatttattag tagattttga ttcaaaggat 2340
attgttgata aatataaagg gaaaaaagta gacttgtatg gtgcttatta tggttatcaa 2400
tgtgcgggtg gtacaccaaa caaaacagct tgtatgtatg gtggtgtaac gttacatgat 2460
aataatcgat tgaccgaaga gaaaaaagtg ccgatcaatt tatggctaga cggtaaacaa 2520
aatacagtac ctttggaaac ggttaaaacg aataagaaaa atgtaactgt tcaggagttg 2580
gatcttcaag caagacgtta tttacaggaa aaatataatt tatataactc tgatgttttt 2640
gatgggaagg ttcagagggg attaatcgtg tttcatactt ctacagaacc ttcggttaat 2700
tacgatttat ttggtgctca aggacagtat tcaaatacac tattaagaat atatagagat 2760
aataaaacga ttaactctga aaacatgcat attgatatat atttatatac aagt 2814
<210> 16
<211> 938
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu
115 120 125
Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr Tyr Ile His Trp Val Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Arg Ile Tyr Pro Thr
165 170 175
Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile
180 185 190
Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Trp Gly Gly Asp
210 215 220
Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
225 230 235 240
Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His Val Asp Lys Leu Ser Gly Gly Gly
690 695 700
Gly Ser Glu Lys Ser Glu Glu Ile Asn Glu Lys Asp Leu Arg Lys Lys
705 710 715 720
Ser Glu Leu Gln Gly Thr Ala Leu Gly Asn Leu Lys Gln Ile Tyr Tyr
725 730 735
Tyr Asn Glu Lys Ala Lys Thr Glu Asn Lys Glu Ser His Asp Gln Phe
740 745 750
Leu Gln His Thr Ile Leu Phe Lys Gly Phe Phe Thr Asp His Ser Trp
755 760 765
Tyr Asn Asp Leu Leu Val Asp Phe Asp Ser Lys Asp Ile Val Asp Lys
770 775 780
Tyr Lys Gly Lys Lys Val Asp Leu Tyr Gly Ala Tyr Tyr Gly Tyr Gln
785 790 795 800
Cys Ala Gly Gly Thr Pro Asn Lys Thr Ala Cys Met Tyr Gly Gly Val
805 810 815
Thr Leu His Asp Asn Asn Arg Leu Thr Glu Glu Lys Lys Val Pro Ile
820 825 830
Asn Leu Trp Leu Asp Gly Lys Gln Asn Thr Val Pro Leu Glu Thr Val
835 840 845
Lys Thr Asn Lys Lys Asn Val Thr Val Gln Glu Leu Asp Leu Gln Ala
850 855 860
Arg Arg Tyr Leu Gln Glu Lys Tyr Asn Leu Tyr Asn Ser Asp Val Phe
865 870 875 880
Asp Gly Lys Val Gln Arg Gly Leu Ile Val Phe His Thr Ser Thr Glu
885 890 895
Pro Ser Val Asn Tyr Asp Leu Phe Gly Ala Gln Gly Gln Tyr Ser Asn
900 905 910
Thr Leu Leu Arg Ile Tyr Arg Asp Asn Lys Thr Ile Asn Ser Glu Asn
915 920 925
Met His Ile Asp Ile Tyr Leu Tyr Thr Ser
930 935
<210> 17
<211> 726
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
caaattgttc tctcccagtc tccagcaatc ctgtcagctt ctcctggaga gaaggtgact 60
atgacttgca gggcttcttc atctgtttct tacatccact ggttccagca gaagcctggt 120
tcttcaccta agccttggat ctacgctaca tctaacttgg catctggagt gcctgtgagg 180
ttctctggtt ctggttcagg tacttcttac tctttgacaa tctctagggt ggaggctgag 240
gacgctgcta cttactactg ccagcagtgg acatctaacc ctccaacatt cggaggtggt 300
actaagttgg agatcaaggg aggtggaggt tctggtggag gaggatctgg aggtggtggt 360
tctcaggtgc agttgcaaca gcctggagct gagttggtga agcctggtgc ttctgtgaag 420
atgtcttgta aggcttctgg atacacattc acttcttaca acatgcactg ggtgaagcag 480
actcctggta ggggtttgga gtggatcgga gctatctacc caggaaacgg agacacatct 540
tacaaccaga agttcaaggg taaggctaca ttgactgctg acaagtcttc atctactgct 600
tacatgcaat tgtcttcttt gacatctgag gactctgcag tttactactg cgctaggtct 660
acatactacg gaggtgactg gtacttcaac gtgtggggag caggtaccac ggtcactgtc 720
tctgca 726
<210> 18
<211> 242
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile
20 25 30
His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Pro
115 120 125
Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys Lys
130 135 140
Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys Gln
145 150 155 160
Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn
165 170 175
Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly
210 215 220
Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr Val
225 230 235 240
Ser Ala
<210> 19
<211> 2088
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
caaattgttc tctcccagtc tccagcaatc ctgtcagctt ctcctggaga gaaggtgact 60
atgacttgca gggcttcttc atctgtttct tacatccact ggttccagca gaagcctggt 120
tcttcaccta agccttggat ctacgctaca tctaacttgg catctggagt gcctgtgagg 180
ttctctggtt ctggttcagg tacttcttac tctttgacaa tctctagggt ggaggctgag 240
gacgctgcta cttactactg ccagcagtgg acatctaacc ctccaacatt cggaggtggt 300
actaagttgg agatcaaggg aggtggaggt tctggtggag gaggatctgg aggtggtggt 360
tctcaggtgc agttgcaaca gcctggagct gagttggtga agcctggtgc ttctgtgaag 420
atgtcttgta aggcttctgg atacacattc acttcttaca acatgcactg ggtgaagcag 480
actcctggta ggggtttgga gtggatcgga gctatctacc caggaaacgg agacacatct 540
tacaaccaga agttcaaggg taaggctaca ttgactgctg acaagtcttc atctactgct 600
tacatgcaat tgtcttcttt gacatctgag gactctgcag tttactactg cgctaggtct 660
acatactacg gaggtgactg gtacttcaac gtgtggggag caggtaccac ggtcactgtc 720
tctgcaggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccac 2088
<210> 20
<211> 696
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile
20 25 30
His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Pro
115 120 125
Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys Lys
130 135 140
Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys Gln
145 150 155 160
Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn
165 170 175
Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly
210 215 220
Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr Val
225 230 235 240
Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His
690 695
<210> 21
<211> 2814
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
caaattgttc tctcccagtc tccagcaatc ctgtcagctt ctcctggaga gaaggtgact 60
atgacttgca gggcttcttc atctgtttct tacatccact ggttccagca gaagcctggt 120
tcttcaccta agccttggat ctacgctaca tctaacttgg catctggagt gcctgtgagg 180
ttctctggtt ctggttcagg tacttcttac tctttgacaa tctctagggt ggaggctgag 240
gacgctgcta cttactactg ccagcagtgg acatctaacc ctccaacatt cggaggtggt 300
actaagttgg agatcaaggg aggtggaggt tctggtggag gaggatctgg aggtggtggt 360
tctcaggtgc agttgcaaca gcctggagct gagttggtga agcctggtgc ttctgtgaag 420
atgtcttgta aggcttctgg atacacattc acttcttaca acatgcactg ggtgaagcag 480
actcctggta ggggtttgga gtggatcgga gctatctacc caggaaacgg agacacatct 540
tacaaccaga agttcaaggg taaggctaca ttgactgctg acaagtcttc atctactgct 600
tacatgcaat tgtcttcttt gacatctgag gactctgcag tttactactg cgctaggtct 660
acatactacg gaggtgactg gtacttcaac gtgtggggag caggtaccac ggtcactgtc 720
tctgcaggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccacgt cgacaagctt 2100
tccggcggag gtggcagcga gaaaagcgaa gaaataaatg aaaaagattt gcgaaaaaag 2160
tctgaattgc agggaacagc tttaggcaat cttaaacaaa tctattatta caatgaaaaa 2220
gctaaaactg aaaataaaga gagtcacgat caatttttac agcatactat attgtttaaa 2280
ggctttttta cagatcattc gtggtataac gatttattag tagattttga ttcaaaggat 2340
attgttgata aatataaagg gaaaaaagta gacttgtatg gtgcttatta tggttatcaa 2400
tgtgcgggtg gtacaccaaa caaaacagct tgtatgtatg gtggtgtaac gttacatgat 2460
aataatcgat tgaccgaaga gaaaaaagtg ccgatcaatt tatggctaga cggtaaacaa 2520
aatacagtac ctttggaaac ggttaaaacg aataagaaaa atgtaactgt tcaggagttg 2580
gatcttcaag caagacgtta tttacaggaa aaatataatt tatataactc tgatgttttt 2640
gatgggaagg ttcagagggg attaatcgtg tttcatactt ctacagaacc ttcggttaat 2700
tacgatttat ttggtgctca aggacagtat tcaaatacac tattaagaat atatagagat 2760
aataaaacga ttaactctga aaacatgcat attgatatat atttatatac aagt 2814
<210> 22
<211> 938
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile
20 25 30
His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Pro
115 120 125
Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Met Ser Cys Lys
130 135 140
Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His Trp Val Lys Gln
145 150 155 160
Thr Pro Gly Arg Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asn
165 170 175
Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Gly
210 215 220
Gly Asp Trp Tyr Phe Asn Val Trp Gly Ala Gly Thr Thr Val Thr Val
225 230 235 240
Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His Val Asp Lys Leu Ser Gly Gly Gly
690 695 700
Gly Ser Glu Lys Ser Glu Glu Ile Asn Glu Lys Asp Leu Arg Lys Lys
705 710 715 720
Ser Glu Leu Gln Gly Thr Ala Leu Gly Asn Leu Lys Gln Ile Tyr Tyr
725 730 735
Tyr Asn Glu Lys Ala Lys Thr Glu Asn Lys Glu Ser His Asp Gln Phe
740 745 750
Leu Gln His Thr Ile Leu Phe Lys Gly Phe Phe Thr Asp His Ser Trp
755 760 765
Tyr Asn Asp Leu Leu Val Asp Phe Asp Ser Lys Asp Ile Val Asp Lys
770 775 780
Tyr Lys Gly Lys Lys Val Asp Leu Tyr Gly Ala Tyr Tyr Gly Tyr Gln
785 790 795 800
Cys Ala Gly Gly Thr Pro Asn Lys Thr Ala Cys Met Tyr Gly Gly Val
805 810 815
Thr Leu His Asp Asn Asn Arg Leu Thr Glu Glu Lys Lys Val Pro Ile
820 825 830
Asn Leu Trp Leu Asp Gly Lys Gln Asn Thr Val Pro Leu Glu Thr Val
835 840 845
Lys Thr Asn Lys Lys Asn Val Thr Val Gln Glu Leu Asp Leu Gln Ala
850 855 860
Arg Arg Tyr Leu Gln Glu Lys Tyr Asn Leu Tyr Asn Ser Asp Val Phe
865 870 875 880
Asp Gly Lys Val Gln Arg Gly Leu Ile Val Phe His Thr Ser Thr Glu
885 890 895
Pro Ser Val Asn Tyr Asp Leu Phe Gly Ala Gln Gly Gln Tyr Ser Asn
900 905 910
Thr Leu Leu Arg Ile Tyr Arg Asp Asn Lys Thr Ile Asn Ser Glu Asn
915 920 925
Met His Ile Asp Ile Tyr Leu Tyr Thr Ser
930 935
<210> 23
<211> 726
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
gatattctgc tgacccagag cccggtgatc ctgagtgttt ccccgggcga acgtgtgtca 60
ttttcgtgtc gcgcgagcca gtctattggt accaatatcc actggtatca gcaacgtacg 120
aacggctctc cgcgcctgct gattaaatac gccagtgaat ccatttcagg catcccgagc 180
cgcttttcgg gcagcggttc tggcaccgat ttcacgctga gtattaactc cgtggaatca 240
gaagatatcg cagactatta ctgccagcaa aacaataact ggccgaccac gtttggtgct 300
ggcaccaaac tggaactgaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctcaag tccaactgaa acaatcgggt ccgggtctgg tccaaccgtc ccaatcactg 420
agcatcacct gtaccgtgtc gggcttctcg ctgaccaatt atggtgtgca ttgggttcgt 480
cagagtccgg gcaaaggtct ggaatggctg ggcgttattt ggtccggcgg taataccgat 540
tacaacaccc cgtttacgag tcgcctgtcc atcaataaag acaactcgaa aagccaggtg 600
tttttcaaaa tgaattcact gcaatcgaac gataccgcga tttattactg cgcacgtgct 660
ctgacgtatt acgactatga atttgcctac tggggccagg gtaccctggt gacggttagc 720
gcggcc 726
<210> 24
<211> 242
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn
20 25 30
Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile
35 40 45
Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser
65 70 75 80
Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr
85 90 95
Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Lys Gln
115 120 125
Ser Gly Pro Gly Leu Val Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys
130 135 140
Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr Gly Val His Trp Val Arg
145 150 155 160
Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Ser Gly
165 170 175
Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn
180 185 190
Lys Asp Asn Ser Lys Ser Gln Val Phe Phe Lys Met Asn Ser Leu Gln
195 200 205
Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr
210 215 220
Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
225 230 235 240
Ala Ala
<210> 25
<211> 2088
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
gatattctgc tgacccagag cccggtgatc ctgagtgttt ccccgggcga acgtgtgtca 60
ttttcgtgtc gcgcgagcca gtctattggt accaatatcc actggtatca gcaacgtacg 120
aacggctctc cgcgcctgct gattaaatac gccagtgaat ccatttcagg catcccgagc 180
cgcttttcgg gcagcggttc tggcaccgat ttcacgctga gtattaactc cgtggaatca 240
gaagatatcg cagactatta ctgccagcaa aacaataact ggccgaccac gtttggtgct 300
ggcaccaaac tggaactgaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctcaag tccaactgaa acaatcgggt ccgggtctgg tccaaccgtc ccaatcactg 420
agcatcacct gtaccgtgtc gggcttctcg ctgaccaatt atggtgtgca ttgggttcgt 480
cagagtccgg gcaaaggtct ggaatggctg ggcgttattt ggtccggcgg taataccgat 540
tacaacaccc cgtttacgag tcgcctgtcc atcaataaag acaactcgaa aagccaggtg 600
tttttcaaaa tgaattcact gcaatcgaac gataccgcga tttattactg cgcacgtgct 660
ctgacgtatt acgactatga atttgcctac tggggccagg gtaccctggt gacggttagc 720
gcggccggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccac 2088
<210> 26
<211> 696
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn
20 25 30
Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile
35 40 45
Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser
65 70 75 80
Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr
85 90 95
Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Lys Gln
115 120 125
Ser Gly Pro Gly Leu Val Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys
130 135 140
Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr Gly Val His Trp Val Arg
145 150 155 160
Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Ser Gly
165 170 175
Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn
180 185 190
Lys Asp Asn Ser Lys Ser Gln Val Phe Phe Lys Met Asn Ser Leu Gln
195 200 205
Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr
210 215 220
Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
225 230 235 240
Ala Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His
690 695
<210> 27
<211> 2814
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
gatattctgc tgacccagag cccggtgatc ctgagtgttt ccccgggcga acgtgtgtca 60
ttttcgtgtc gcgcgagcca gtctattggt accaatatcc actggtatca gcaacgtacg 120
aacggctctc cgcgcctgct gattaaatac gccagtgaat ccatttcagg catcccgagc 180
cgcttttcgg gcagcggttc tggcaccgat ttcacgctga gtattaactc cgtggaatca 240
gaagatatcg cagactatta ctgccagcaa aacaataact ggccgaccac gtttggtgct 300
ggcaccaaac tggaactgaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctcaag tccaactgaa acaatcgggt ccgggtctgg tccaaccgtc ccaatcactg 420
agcatcacct gtaccgtgtc gggcttctcg ctgaccaatt atggtgtgca ttgggttcgt 480
cagagtccgg gcaaaggtct ggaatggctg ggcgttattt ggtccggcgg taataccgat 540
tacaacaccc cgtttacgag tcgcctgtcc atcaataaag acaactcgaa aagccaggtg 600
tttttcaaaa tgaattcact gcaatcgaac gataccgcga tttattactg cgcacgtgct 660
ctgacgtatt acgactatga atttgcctac tggggccagg gtaccctggt gacggttagc 720
gcggccggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccacgt cgacaagctt 2100
tccggcggag gtggcagcga gaaaagcgaa gaaataaatg aaaaagattt gcgaaaaaag 2160
tctgaattgc agggaacagc tttaggcaat cttaaacaaa tctattatta caatgaaaaa 2220
gctaaaactg aaaataaaga gagtcacgat caatttttac agcatactat attgtttaaa 2280
ggctttttta cagatcattc gtggtataac gatttattag tagattttga ttcaaaggat 2340
attgttgata aatataaagg gaaaaaagta gacttgtatg gtgcttatta tggttatcaa 2400
tgtgcgggtg gtacaccaaa caaaacagct tgtatgtatg gtggtgtaac gttacatgat 2460
aataatcgat tgaccgaaga gaaaaaagtg ccgatcaatt tatggctaga cggtaaacaa 2520
aatacagtac ctttggaaac ggttaaaacg aataagaaaa atgtaactgt tcaggagttg 2580
gatcttcaag caagacgtta tttacaggaa aaatataatt tatataactc tgatgttttt 2640
gatgggaagg ttcagagggg attaatcgtg tttcatactt ctacagaacc ttcggttaat 2700
tacgatttat ttggtgctca aggacagtat tcaaatacac tattaagaat atatagagat 2760
aataaaacga ttaactctga aaacatgcat attgatatat atttatatac aagt 2814
<210> 28
<211> 938
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn
20 25 30
Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile
35 40 45
Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser
65 70 75 80
Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr
85 90 95
Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Lys Gln
115 120 125
Ser Gly Pro Gly Leu Val Gln Pro Ser Gln Ser Leu Ser Ile Thr Cys
130 135 140
Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr Gly Val His Trp Val Arg
145 150 155 160
Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Ser Gly
165 170 175
Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr Ser Arg Leu Ser Ile Asn
180 185 190
Lys Asp Asn Ser Lys Ser Gln Val Phe Phe Lys Met Asn Ser Leu Gln
195 200 205
Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala Arg Ala Leu Thr Tyr Tyr
210 215 220
Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
225 230 235 240
Ala Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His Val Asp Lys Leu Ser Gly Gly Gly
690 695 700
Gly Ser Glu Lys Ser Glu Glu Ile Asn Glu Lys Asp Leu Arg Lys Lys
705 710 715 720
Ser Glu Leu Gln Gly Thr Ala Leu Gly Asn Leu Lys Gln Ile Tyr Tyr
725 730 735
Tyr Asn Glu Lys Ala Lys Thr Glu Asn Lys Glu Ser His Asp Gln Phe
740 745 750
Leu Gln His Thr Ile Leu Phe Lys Gly Phe Phe Thr Asp His Ser Trp
755 760 765
Tyr Asn Asp Leu Leu Val Asp Phe Asp Ser Lys Asp Ile Val Asp Lys
770 775 780
Tyr Lys Gly Lys Lys Val Asp Leu Tyr Gly Ala Tyr Tyr Gly Tyr Gln
785 790 795 800
Cys Ala Gly Gly Thr Pro Asn Lys Thr Ala Cys Met Tyr Gly Gly Val
805 810 815
Thr Leu His Asp Asn Asn Arg Leu Thr Glu Glu Lys Lys Val Pro Ile
820 825 830
Asn Leu Trp Leu Asp Gly Lys Gln Asn Thr Val Pro Leu Glu Thr Val
835 840 845
Lys Thr Asn Lys Lys Asn Val Thr Val Gln Glu Leu Asp Leu Gln Ala
850 855 860
Arg Arg Tyr Leu Gln Glu Lys Tyr Asn Leu Tyr Asn Ser Asp Val Phe
865 870 875 880
Asp Gly Lys Val Gln Arg Gly Leu Ile Val Phe His Thr Ser Thr Glu
885 890 895
Pro Ser Val Asn Tyr Asp Leu Phe Gly Ala Gln Gly Gln Tyr Ser Asn
900 905 910
Thr Leu Leu Arg Ile Tyr Arg Asp Asn Lys Thr Ile Asn Ser Glu Asn
915 920 925
Met His Ile Asp Ile Tyr Leu Tyr Thr Ser
930 935
<210> 29
<211> 726
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60
atcacttgcc aggcgagtca ggacattagc aactatttaa attggtatca gcagaaacca 120
gggaaagccc ctaagctcct gatctacgat gcatccaatt tggaaacagg ggtcccatca 180
aggttcagtg gaagtggatc tgggacagat tttactttca ccatcagcag cctgcagcct 240
gaagatattg caacatattt ctgtcaacac tttgatcatc tccctctcgc tttcggcgga 300
gggaccaagg tggagatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctcagg tgcagctgca ggagtcgggc ccaggactgg tgaagccttc ggagaccctg 420
tccctcacct gcactgtctc tggtggctcc gtcagtagtg gtgactacta ctggacctgg 480
atccggcagt ccccagggaa gggactggag tggattgggc atatctatta cagtgggaac 540
accaactaca acccctccct caagagtcga ctcaccatat caatagacac gtccaagacc 600
cagttctccc tgaagctgag ctctgtgacc gctgcggaca cggccatcta ttactgtgtg 660
agggaccgag taactggtgc ttttgatatc tggggccaag ggacaatggt caccgtctct 720
tcagcg 726
<210> 30
<211> 242
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln His Phe Asp His Leu Pro Leu
85 90 95
Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu
115 120 125
Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys
130 135 140
Thr Val Ser Gly Gly Ser Val Ser Ser Gly Asp Tyr Tyr Trp Thr Trp
145 150 155 160
Ile Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Ile Gly His Ile Tyr
165 170 175
Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Leu Thr
180 185 190
Ile Ser Ile Asp Thr Ser Lys Thr Gln Phe Ser Leu Lys Leu Ser Ser
195 200 205
Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Val Arg Asp Arg Val
210 215 220
Thr Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser
225 230 235 240
Ser Ala
<210> 31
<211> 2088
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60
atcacttgcc aggcgagtca ggacattagc aactatttaa attggtatca gcagaaacca 120
gggaaagccc ctaagctcct gatctacgat gcatccaatt tggaaacagg ggtcccatca 180
aggttcagtg gaagtggatc tgggacagat tttactttca ccatcagcag cctgcagcct 240
gaagatattg caacatattt ctgtcaacac tttgatcatc tccctctcgc tttcggcgga 300
gggaccaagg tggagatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctcagg tgcagctgca ggagtcgggc ccaggactgg tgaagccttc ggagaccctg 420
tccctcacct gcactgtctc tggtggctcc gtcagtagtg gtgactacta ctggacctgg 480
atccggcagt ccccagggaa gggactggag tggattgggc atatctatta cagtgggaac 540
accaactaca acccctccct caagagtcga ctcaccatat caatagacac gtccaagacc 600
cagttctccc tgaagctgag ctctgtgacc gctgcggaca cggccatcta ttactgtgtg 660
agggaccgag taactggtgc ttttgatatc tggggccaag ggacaatggt caccgtctct 720
tcagcgggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccac 2088
<210> 32
<211> 696
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln His Phe Asp His Leu Pro Leu
85 90 95
Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu
115 120 125
Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys
130 135 140
Thr Val Ser Gly Gly Ser Val Ser Ser Gly Asp Tyr Tyr Trp Thr Trp
145 150 155 160
Ile Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Ile Gly His Ile Tyr
165 170 175
Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Leu Thr
180 185 190
Ile Ser Ile Asp Thr Ser Lys Thr Gln Phe Ser Leu Lys Leu Ser Ser
195 200 205
Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Val Arg Asp Arg Val
210 215 220
Thr Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser
225 230 235 240
Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His
690 695
<210> 33
<211> 2814
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60
atcacttgcc aggcgagtca ggacattagc aactatttaa attggtatca gcagaaacca 120
gggaaagccc ctaagctcct gatctacgat gcatccaatt tggaaacagg ggtcccatca 180
aggttcagtg gaagtggatc tgggacagat tttactttca ccatcagcag cctgcagcct 240
gaagatattg caacatattt ctgtcaacac tttgatcatc tccctctcgc tttcggcgga 300
gggaccaagg tggagatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctcagg tgcagctgca ggagtcgggc ccaggactgg tgaagccttc ggagaccctg 420
tccctcacct gcactgtctc tggtggctcc gtcagtagtg gtgactacta ctggacctgg 480
atccggcagt ccccagggaa gggactggag tggattgggc atatctatta cagtgggaac 540
accaactaca acccctccct caagagtcga ctcaccatat caatagacac gtccaagacc 600
cagttctccc tgaagctgag ctctgtgacc gctgcggaca cggccatcta ttactgtgtg 660
agggaccgag taactggtgc ttttgatatc tggggccaag ggacaatggt caccgtctct 720
tcagcgggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccacgt cgacaagctt 2100
tccggcggag gtggcagcga gaaaagcgaa gaaataaatg aaaaagattt gcgaaaaaag 2160
tctgaattgc agggaacagc tttaggcaat cttaaacaaa tctattatta caatgaaaaa 2220
gctaaaactg aaaataaaga gagtcacgat caatttttac agcatactat attgtttaaa 2280
ggctttttta cagatcattc gtggtataac gatttattag tagattttga ttcaaaggat 2340
attgttgata aatataaagg gaaaaaagta gacttgtatg gtgcttatta tggttatcaa 2400
tgtgcgggtg gtacaccaaa caaaacagct tgtatgtatg gtggtgtaac gttacatgat 2460
aataatcgat tgaccgaaga gaaaaaagtg ccgatcaatt tatggctaga cggtaaacaa 2520
aatacagtac ctttggaaac ggttaaaacg aataagaaaa atgtaactgt tcaggagttg 2580
gatcttcaag caagacgtta tttacaggaa aaatataatt tatataactc tgatgttttt 2640
gatgggaagg ttcagagggg attaatcgtg tttcatactt ctacagaacc ttcggttaat 2700
tacgatttat ttggtgctca aggacagtat tcaaatacac tattaagaat atatagagat 2760
aataaaacga ttaactctga aaacatgcat attgatatat atttatatac aagt 2814
<210> 34
<211> 938
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln His Phe Asp His Leu Pro Leu
85 90 95
Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Glu
115 120 125
Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys
130 135 140
Thr Val Ser Gly Gly Ser Val Ser Ser Gly Asp Tyr Tyr Trp Thr Trp
145 150 155 160
Ile Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Ile Gly His Ile Tyr
165 170 175
Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Leu Thr
180 185 190
Ile Ser Ile Asp Thr Ser Lys Thr Gln Phe Ser Leu Lys Leu Ser Ser
195 200 205
Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Val Arg Asp Arg Val
210 215 220
Thr Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser
225 230 235 240
Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His Val Asp Lys Leu Ser Gly Gly Gly
690 695 700
Gly Ser Glu Lys Ser Glu Glu Ile Asn Glu Lys Asp Leu Arg Lys Lys
705 710 715 720
Ser Glu Leu Gln Gly Thr Ala Leu Gly Asn Leu Lys Gln Ile Tyr Tyr
725 730 735
Tyr Asn Glu Lys Ala Lys Thr Glu Asn Lys Glu Ser His Asp Gln Phe
740 745 750
Leu Gln His Thr Ile Leu Phe Lys Gly Phe Phe Thr Asp His Ser Trp
755 760 765
Tyr Asn Asp Leu Leu Val Asp Phe Asp Ser Lys Asp Ile Val Asp Lys
770 775 780
Tyr Lys Gly Lys Lys Val Asp Leu Tyr Gly Ala Tyr Tyr Gly Tyr Gln
785 790 795 800
Cys Ala Gly Gly Thr Pro Asn Lys Thr Ala Cys Met Tyr Gly Gly Val
805 810 815
Thr Leu His Asp Asn Asn Arg Leu Thr Glu Glu Lys Lys Val Pro Ile
820 825 830
Asn Leu Trp Leu Asp Gly Lys Gln Asn Thr Val Pro Leu Glu Thr Val
835 840 845
Lys Thr Asn Lys Lys Asn Val Thr Val Gln Glu Leu Asp Leu Gln Ala
850 855 860
Arg Arg Tyr Leu Gln Glu Lys Tyr Asn Leu Tyr Asn Ser Asp Val Phe
865 870 875 880
Asp Gly Lys Val Gln Arg Gly Leu Ile Val Phe His Thr Ser Thr Glu
885 890 895
Pro Ser Val Asn Tyr Asp Leu Phe Gly Ala Gln Gly Gln Tyr Ser Asn
900 905 910
Thr Leu Leu Arg Ile Tyr Arg Asp Asn Lys Thr Ile Asn Ser Glu Asn
915 920 925
Met His Ile Asp Ile Tyr Leu Tyr Thr Ser
930 935
<210> 35
<211> 726
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
gacatccaga tgacccagtc tccatcctcc gtgtctgcat ctatagggga cagagtcacc 60
atcacttgtc gggcgagtca gggtattgac aactggttag gctggtatca gcagaagcca 120
gggaaagccc ctaagctcct gatctatgat gcatccaatt tggacactgg ggtcccatca 180
aggttcagcg gcagtggatc tgggacatat ttcactctca ctatcagcag cctgcaggct 240
gaagactttg cagtttactt ttgtcaacag gctaaggctt tcccccccac tttcggcgga 300
gggaccaagg tggacatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctgagg tgcagctggt gcagtctggg ggaggcctgg tcaagcctgg ggggtccctg 420
agactctcct gtgcagcctc tggattcacc ttcagtagct atagcatgaa ctgggtccgc 480
caggctccag ggaaggggct ggagtgggtc tcatccatta gtagtagtag tagttacata 540
tactacgcag actcagtgaa gggccgattc accatctcca gagacaacgc caagaactca 600
ctgtatctgc aaatgaacag cctgagagcc gaggacacgg ctgtgtatta ctgtgcgaga 660
gttaccgatg cttttgatat ctggggccaa gggacaatgg tcaccgtctc ttcagcttcc 720
acgaaa 726
<210> 36
<211> 242
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Ile Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asp Asn Trp
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Leu Asp Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Tyr Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ala Lys Ala Phe Pro Pro
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Gln
115 120 125
Ser Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ser Met Asn Trp Val Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Ser Ser
165 170 175
Ser Ser Tyr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile
180 185 190
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Thr Asp Ala
210 215 220
Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser
225 230 235 240
Thr Lys
<210> 37
<211> 2088
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
gacatccaga tgacccagtc tccatcctcc gtgtctgcat ctatagggga cagagtcacc 60
atcacttgtc gggcgagtca gggtattgac aactggttag gctggtatca gcagaagcca 120
gggaaagccc ctaagctcct gatctatgat gcatccaatt tggacactgg ggtcccatca 180
aggttcagcg gcagtggatc tgggacatat ttcactctca ctatcagcag cctgcaggct 240
gaagactttg cagtttactt ttgtcaacag gctaaggctt tcccccccac tttcggcgga 300
gggaccaagg tggacatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctgagg tgcagctggt gcagtctggg ggaggcctgg tcaagcctgg ggggtccctg 420
agactctcct gtgcagcctc tggattcacc ttcagtagct atagcatgaa ctgggtccgc 480
caggctccag ggaaggggct ggagtgggtc tcatccatta gtagtagtag tagttacata 540
tactacgcag actcagtgaa gggccgattc accatctcca gagacaacgc caagaactca 600
ctgtatctgc aaatgaacag cctgagagcc gaggacacgg ctgtgtatta ctgtgcgaga 660
gttaccgatg cttttgatat ctggggccaa gggacaatgg tcaccgtctc ttcagcttcc 720
acgaaaggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccac 2088
<210> 38
<211> 696
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Ile Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asp Asn Trp
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Leu Asp Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Tyr Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ala Lys Ala Phe Pro Pro
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Gln
115 120 125
Ser Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ser Met Asn Trp Val Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Ser Ser
165 170 175
Ser Ser Tyr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile
180 185 190
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Thr Asp Ala
210 215 220
Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser
225 230 235 240
Thr Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His
690 695
<210> 39
<211> 2814
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
gacatccaga tgacccagtc tccatcctcc gtgtctgcat ctatagggga cagagtcacc 60
atcacttgtc gggcgagtca gggtattgac aactggttag gctggtatca gcagaagcca 120
gggaaagccc ctaagctcct gatctatgat gcatccaatt tggacactgg ggtcccatca 180
aggttcagcg gcagtggatc tgggacatat ttcactctca ctatcagcag cctgcaggct 240
gaagactttg cagtttactt ttgtcaacag gctaaggctt tcccccccac tttcggcgga 300
gggaccaagg tggacatcaa aggaggtgga ggttctggtg gaggaggatc tggaggtggt 360
ggttctgagg tgcagctggt gcagtctggg ggaggcctgg tcaagcctgg ggggtccctg 420
agactctcct gtgcagcctc tggattcacc ttcagtagct atagcatgaa ctgggtccgc 480
caggctccag ggaaggggct ggagtgggtc tcatccatta gtagtagtag tagttacata 540
tactacgcag actcagtgaa gggccgattc accatctcca gagacaacgc caagaactca 600
ctgtatctgc aaatgaacag cctgagagcc gaggacacgg ctgtgtatta ctgtgcgaga 660
gttaccgatg cttttgatat ctggggccaa gggacaatgg tcaccgtctc ttcagcttcc 720
acgaaaggag gtggaggttc tggtggagga ggatctggag gtggtggttc tgttatccac 780
gtgaccaagg aagtgaaaga agtggcaacg ctgtcctgtg gtcacaatgt ttctgttgaa 840
gagctggcac aaactcgcat ctactggcaa aaggagaaga aaatggtgct gactatgatg 900
tctggggaca tgaatatatg gcccgagtac aagaaccgga ccatctttga tatcactaat 960
aacctctcca ttgtgatcct ggctctgcgc ccatctgacg agggcacata cgagtgtgtt 1020
gttctgaagt atgaaaaaga cgctttcaag cgggaacacc tggctgaagt gacgttatca 1080
gtcaaagctg acttccctac acctagtata tctgactttg aaattccaac ttctaatatt 1140
agaaggataa tttgctcaac ctctggaggt tttccagagc ctcacctctc ctggttggaa 1200
aatggagaag aattaaatgc catcaacaca acagtttccc aagatcctga aactgagctc 1260
tatgctgtta gcagcaaact ggatttcaat atgacaacca accacagctt catgtgtctc 1320
atcaagtatg gacatttaag agtgaatcag accttcaact ggaatacaac caagcaagag 1380
cattttcctg ataacgtcga caagctttcc ggcggaggtg gcgctcctct gaagattcaa 1440
gcttatttca atgagactgc agacctgcca tgccaatttg caaactctca aaaccaaagc 1500
ctgagtgagc tagtagtatt ttggcaggac caggaaaact tggttctgaa tgaggtatac 1560
ttaggcaaag agaaatttga cagtgttcat tccaagtata tgggccgcac aagttttgat 1620
tcggacagtt ggaccctgag acttcacaat cttcagatca aggacaaggg cttgtatcaa 1680
tgtatcatcc atcacaaaaa gcccacagga atgattcgca tccaccagat gaattctgaa 1740
ctgtcagtgc ttgctaactt cagtcaacct gaaatagtac caatttctaa tataacagaa 1800
aatgtgtaca taaatttgac ctgctcatct atacacggtt acccagaacc taagaagatg 1860
agtgttttgc taagaaccaa gaattcaact atcgagtatg atggtattat gcagaaatct 1920
caagataatg tcacagaact gtacgacgtt tccatcagct tgtctgtttc attccctgat 1980
gttacgagca atatgaccat cttctgtatt ctggaaactg acaagacgcg gcttttatct 2040
tcacctttct ctatagagct tgaggaccct cagcctcccc cagaccacgt cgacaagctt 2100
tccggcggag gtggcagcga gaaaagcgaa gaaataaatg aaaaagattt gcgaaaaaag 2160
tctgaattgc agggaacagc tttaggcaat cttaaacaaa tctattatta caatgaaaaa 2220
gctaaaactg aaaataaaga gagtcacgat caatttttac agcatactat attgtttaaa 2280
ggctttttta cagatcattc gtggtataac gatttattag tagattttga ttcaaaggat 2340
attgttgata aatataaagg gaaaaaagta gacttgtatg gtgcttatta tggttatcaa 2400
tgtgcgggtg gtacaccaaa caaaacagct tgtatgtatg gtggtgtaac gttacatgat 2460
aataatcgat tgaccgaaga gaaaaaagtg ccgatcaatt tatggctaga cggtaaacaa 2520
aatacagtac ctttggaaac ggttaaaacg aataagaaaa atgtaactgt tcaggagttg 2580
gatcttcaag caagacgtta tttacaggaa aaatataatt tatataactc tgatgttttt 2640
gatgggaagg ttcagagggg attaatcgtg tttcatactt ctacagaacc ttcggttaat 2700
tacgatttat ttggtgctca aggacagtat tcaaatacac tattaagaat atatagagat 2760
aataaaacga ttaactctga aaacatgcat attgatatat atttatatac aagt 2814
<210> 40
<211> 938
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Ile Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asp Asn Trp
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Leu Asp Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Tyr Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ala Lys Ala Phe Pro Pro
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Gln
115 120 125
Ser Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ser Met Asn Trp Val Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Ser Ser
165 170 175
Ser Ser Tyr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile
180 185 190
Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Thr Asp Ala
210 215 220
Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser
225 230 235 240
Thr Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Val Ile His Val Thr Lys Glu Val Lys Glu Val Ala Thr Leu Ser
260 265 270
Cys Gly His Asn Val Ser Val Glu Glu Leu Ala Gln Thr Arg Ile Tyr
275 280 285
Trp Gln Lys Glu Lys Lys Met Val Leu Thr Met Met Ser Gly Asp Met
290 295 300
Asn Ile Trp Pro Glu Tyr Lys Asn Arg Thr Ile Phe Asp Ile Thr Asn
305 310 315 320
Asn Leu Ser Ile Val Ile Leu Ala Leu Arg Pro Ser Asp Glu Gly Thr
325 330 335
Tyr Glu Cys Val Val Leu Lys Tyr Glu Lys Asp Ala Phe Lys Arg Glu
340 345 350
His Leu Ala Glu Val Thr Leu Ser Val Lys Ala Asp Phe Pro Thr Pro
355 360 365
Ser Ile Ser Asp Phe Glu Ile Pro Thr Ser Asn Ile Arg Arg Ile Ile
370 375 380
Cys Ser Thr Ser Gly Gly Phe Pro Glu Pro His Leu Ser Trp Leu Glu
385 390 395 400
Asn Gly Glu Glu Leu Asn Ala Ile Asn Thr Thr Val Ser Gln Asp Pro
405 410 415
Glu Thr Glu Leu Tyr Ala Val Ser Ser Lys Leu Asp Phe Asn Met Thr
420 425 430
Thr Asn His Ser Phe Met Cys Leu Ile Lys Tyr Gly His Leu Arg Val
435 440 445
Asn Gln Thr Phe Asn Trp Asn Thr Thr Lys Gln Glu His Phe Pro Asp
450 455 460
Asn Val Asp Lys Leu Ser Gly Gly Gly Gly Ala Pro Leu Lys Ile Gln
465 470 475 480
Ala Tyr Phe Asn Glu Thr Ala Asp Leu Pro Cys Gln Phe Ala Asn Ser
485 490 495
Gln Asn Gln Ser Leu Ser Glu Leu Val Val Phe Trp Gln Asp Gln Glu
500 505 510
Asn Leu Val Leu Asn Glu Val Tyr Leu Gly Lys Glu Lys Phe Asp Ser
515 520 525
Val His Ser Lys Tyr Met Gly Arg Thr Ser Phe Asp Ser Asp Ser Trp
530 535 540
Thr Leu Arg Leu His Asn Leu Gln Ile Lys Asp Lys Gly Leu Tyr Gln
545 550 555 560
Cys Ile Ile His His Lys Lys Pro Thr Gly Met Ile Arg Ile His Gln
565 570 575
Met Asn Ser Glu Leu Ser Val Leu Ala Asn Phe Ser Gln Pro Glu Ile
580 585 590
Val Pro Ile Ser Asn Ile Thr Glu Asn Val Tyr Ile Asn Leu Thr Cys
595 600 605
Ser Ser Ile His Gly Tyr Pro Glu Pro Lys Lys Met Ser Val Leu Leu
610 615 620
Arg Thr Lys Asn Ser Thr Ile Glu Tyr Asp Gly Ile Met Gln Lys Ser
625 630 635 640
Gln Asp Asn Val Thr Glu Leu Tyr Asp Val Ser Ile Ser Leu Ser Val
645 650 655
Ser Phe Pro Asp Val Thr Ser Asn Met Thr Ile Phe Cys Ile Leu Glu
660 665 670
Thr Asp Lys Thr Arg Leu Leu Ser Ser Pro Phe Ser Ile Glu Leu Glu
675 680 685
Asp Pro Gln Pro Pro Pro Asp His Val Asp Lys Leu Ser Gly Gly Gly
690 695 700
Gly Ser Glu Lys Ser Glu Glu Ile Asn Glu Lys Asp Leu Arg Lys Lys
705 710 715 720
Ser Glu Leu Gln Gly Thr Ala Leu Gly Asn Leu Lys Gln Ile Tyr Tyr
725 730 735
Tyr Asn Glu Lys Ala Lys Thr Glu Asn Lys Glu Ser His Asp Gln Phe
740 745 750
Leu Gln His Thr Ile Leu Phe Lys Gly Phe Phe Thr Asp His Ser Trp
755 760 765
Tyr Asn Asp Leu Leu Val Asp Phe Asp Ser Lys Asp Ile Val Asp Lys
770 775 780
Tyr Lys Gly Lys Lys Val Asp Leu Tyr Gly Ala Tyr Tyr Gly Tyr Gln
785 790 795 800
Cys Ala Gly Gly Thr Pro Asn Lys Thr Ala Cys Met Tyr Gly Gly Val
805 810 815
Thr Leu His Asp Asn Asn Arg Leu Thr Glu Glu Lys Lys Val Pro Ile
820 825 830
Asn Leu Trp Leu Asp Gly Lys Gln Asn Thr Val Pro Leu Glu Thr Val
835 840 845
Lys Thr Asn Lys Lys Asn Val Thr Val Gln Glu Leu Asp Leu Gln Ala
850 855 860
Arg Arg Tyr Leu Gln Glu Lys Tyr Asn Leu Tyr Asn Ser Asp Val Phe
865 870 875 880
Asp Gly Lys Val Gln Arg Gly Leu Ile Val Phe His Thr Ser Thr Glu
885 890 895
Pro Ser Val Asn Tyr Asp Leu Phe Gly Ala Gln Gly Gln Tyr Ser Asn
900 905 910
Thr Leu Leu Arg Ile Tyr Arg Asp Asn Lys Thr Ile Asn Ser Glu Asn
915 920 925
Met His Ile Asp Ile Tyr Leu Tyr Thr Ser
930 935
<210> 41
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
gtcgacaagc tttccggcgg aggtggc 27
<210> 42
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Val Asp Lys Leu Ser Gly Gly Gly Gly
1 5
<210> 43
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
ggaggtggag gttctggtgg aggaggatct ggaggtggtg gttct 45
<210> 44
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15

Claims (6)

1.癌靶向增强型抗肿瘤融合蛋白，其特征在于包括：a)抗体和b)诱导并激活T细胞的融合蛋白；所述抗体为单链抗体形式的Trastuzumab、Rituximab、Cetuximab、Panitumumab或Ramucirumab；
所述Trastuzumab的氨基酸序列用SEQ ID NO.12所示；
所述Rituximab的氨基酸序列用SEQ ID NO.18所示；
所述Cetuximab的氨基酸序列用SEQ ID NO.24所示；
所述Panitumumab的氨基酸序列用SEQ ID NO.30所示；
所述Ramucirumab的氨基酸序列用SEQ ID NO.36所示；
所述诱导并激活T细胞的融合蛋白为诱导并激活T细胞的共刺激分子B7.1、诱导并激活T细胞的共刺激分子B7.2和超抗原金黄色葡萄球菌肠毒素A融合；
所述诱导并激活T细胞的共刺激分子B7.1简称B7.1；所述诱导并激活T细胞的共刺激分子B7.2简称B7.2；超抗原金黄色葡萄球菌肠毒素A简称SEA；
B7.1-B7.2-SEA的氨基酸序列用SEQ ID NO.10所示。
2.根据权利要求1所述的癌靶向增强型抗肿瘤融合蛋白，其特征是所述癌靶向增强型抗肿瘤融合蛋白为：
Trastuzumab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID NO .16所示；
Rituximab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID NO .22所示；
Cetuximab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID NO .28所示；
Panitumumab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID NO .34所示；
Ramucirumab-B7.1-B7.2-SEA融合且氨基酸序列用SEQ ID NO .40所示。
3.重组载体，其特征在于含有编码权利要求2所述癌靶向增强型抗肿瘤融合蛋白的核苷酸序列；
编码SEQ ID No.16的核苷酸序列用SEQ ID NO .15所示；
编码SEQ ID No.22的核苷酸序列用SEQ ID NO .21所示；
编码SEQ ID No.28的核苷酸序列用SEQ ID NO .27所示；
编码SEQ ID No.34的核苷酸序列用SEQ ID NO .33所示；
或编码SEQ ID No.40的核苷酸序列用SEQ ID NO .39所示。
4.宿主细胞，其特征是含有权利要求3的重组载体。
5.癌靶向增强型抗肿瘤融合蛋白的制备方法，其特征是培养权利要求4的宿主细胞，收集癌靶向增强型抗肿瘤融合蛋白。
6.权利要求1或2的癌靶向增强型抗肿瘤融合蛋白在制备治疗癌症和抗实体肿瘤药物的应用。
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