CN116217732A - 靶向cd22和cd19的双特异性抗体及其嵌合抗原受体和应用 - Google Patents

靶向cd22和cd19的双特异性抗体及其嵌合抗原受体和应用 Download PDF

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CN116217732A
CN116217732A CN202111478760.XA CN202111478760A CN116217732A CN 116217732 A CN116217732 A CN 116217732A CN 202111478760 A CN202111478760 A CN 202111478760A CN 116217732 A CN116217732 A CN 116217732A
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黄霞
赵永春
赵文旭
幺瑞娜
沈俊杰
杨智
陈军
洪娟
徐艳敏
齐亚男
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Chongqing Jingzhun Biological Industrial Technology Institute Co ltd
Chongqing Precision Biotech Co ltd
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Abstract

本发明属于细胞工程技术领域，具体涉及一种靶向CD22和CD19的双特异性抗体及其嵌合抗原受体和应用。该双特异性抗体中，CD22轻链与CD22重链通过linker4连接构成CD22单链抗体，CD19轻链与CD19重链通过linker4连接构成CD19单链抗体，其包括以下几种结构中的一种：(1)CD22单链抗体与CD19单链抗体通过linker1或liner2连接；(2)CD19单链抗体的两端通过linker3分别连接CD22轻链和CD22重链。本发明提供的包含双特异性抗体的嵌合抗原受体可同时靶向两个不同的肿瘤抗原，提高对肿瘤细胞杀伤，减少其免疫逃逸的几率，降低CAR‑T治疗后肿瘤复发率。

Description

靶向CD22和CD19的双特异性抗体及其嵌合抗原受体和应用
技术领域
本发明属于细胞工程技术领域，具体涉及一种靶向CD22和CD19的双特异性抗体及其嵌合抗原受体和应用。
背景技术
嵌合抗原受体(chimeric antigen receptor,CAR)修饰的T细胞作为一种免疫治疗策略，在肿瘤治疗中受到广泛的重视和应用。CAR的结构一般由胞外靶向连接区(常为具有抗原识别功能的单链抗体)，铰链区，跨膜区和胞内信号转导区四部分组成。目前根据胞内信号转导区加入共刺激分子的数量，将CAR分为一代(无共刺激分子)、二代(含一种共刺激分子)和三代(含两种共刺激分子)。目前应用最多的是二代CAR。
B细胞恶性肿瘤如B细胞淋巴细胞白血病(B-ALL)和淋巴瘤是B淋巴细胞异常克隆性增殖所致的恶性疾病，尽管目前化疗效果显著，但仍有15％的儿童B-ALL和60％的成人BALL患者因对化疗药物耐药而预后不佳；淋巴瘤患者经一线方案治疗后仍有15％复发，这些患者进行造血干细胞移植后，仍有50％最终复发。因此，寻找有效的细胞免疫治疗手段来治疗B细胞恶性肿瘤一直是血液学界研究热点。
CD19和CD22是B淋巴细胞表面特有的抗原分子。CD19几乎表达于所有的B-ALL和淋巴瘤细胞表面，在B系正常造血细胞表面表达，而在正常非造血组织中没有表达。CD22是一种B系分化抗原，表达于B细胞发育的各个阶段，B细胞分化为浆细胞后不再表达CD22；60％～80％的B细胞恶性肿瘤表达CD22，90％以上的弥漫大B细胞淋巴瘤(DLBCLs)和滤泡淋巴瘤(FLs)为CD22阳性；几乎所有的B前体细胞急性淋巴细胞白血病(B-ALL)表达CD22；慢性淋巴细胞白血病、毛细胞白血病(HCL)也具有CD22表达。与CD19抗原相似，CD22同样是B细胞限制性表达，不表达于其他实质细胞，亦不表达于造血干细胞，因此作为B细胞肿瘤抗原的特异性高，已经成为B细胞恶性肿瘤中的主要治疗靶点。
目前，尽管CD19-CAR-T在难治、复发B-ALL的治疗中取得了较好疗效，但仍有患者在治疗中出现CD19抗原突变或丢失等异常，致使CD19-CAR-T细胞不能识别及杀伤B-ALL细胞，导致疾病复发。CD22与CD19在肿瘤细胞表面具有广泛的共表达，且在CD19抗原丢失后CD22仍然保留，但CD22在肿瘤细胞表面的表达丰度相对较弱。因此，针对CD22抗原需要一个高亲和力的单链抗体，同时CD19、CD22双靶点的CAR-T细胞可以有效避免抗原变异，减少复发，提高抗B系恶性肿瘤的疗效。
发明内容
有鉴于此，针对现有技术中CD19CAR-T治疗期间因B-ALL的逃避机制等而产生复发或无效，本发明目的在于提供一种同时靶向CD22和CD19的双靶点的抗体及嵌合抗原受体。
所述靶向CD22和CD19的双特异性抗体，包括CD22轻链、CD22重链、CD19轻链、CD19重链，所述CD22轻链的可变区包含L-CDR1、L-CDR2和L-CDR3，所述CD22重链的可变区包含H-CDR1、H-CDR2和H-CDR3；
所述L-CDR1、L-CDR2和L-CDR3选自以下氨基酸序列组合中一种：
(1)QSVSSNL、GAS、QQYHSWPPLT；
(2)QSVSSTY、GAS、QQRSNWLT；
(3)QTIDNY、AAS、QQSYSTPPMYT；
(4)QSISSY、SAS、QQSFTTPFT；
(5)QSISSY、AAS、QQYSSYPHT；
所述H-CDR1、H-CDR2和H-CDR选自以下氨基酸序列组合中一种：
(1)GYTFTSYG、ISAYNGNT、ARDFQGIAVADLDY；
(2)GYTFTSYY、INPSGGST、ARGVGSYAMDV；
(3)GFKFDDYP、ISWDGKTT、AKDRSRTGWGYGGMDV；
(4)GDSVSSNSAA、TYYRSKWYN、ARSVGIARWDV；
(5)GDSVSSNSAA、TYYRSKWYN、ARATGTASGWFDP。
优选地，所述CD22轻链包含如SEQ ID NO:1或SEQ ID NO:66所示的氨基酸序列或其功能性变体；所述CD22重链包含如SEQ ID NO:2或SEQ ID NO:67所示的氨基酸序列或其功能性变体。
优选地，所述CD19轻链包含如SEQ ID NO:3或SEQ ID NO:68所示的氨基酸序列或其功能性变体；所述CD19重链包含如SEQ ID NO:4或SEQ ID NO:69所示的氨基酸序列或其功能性变体。
优选地，所述双特异性抗体包括以下几种结构中的一种：
(1)CD22VL-linker4-CD22VH-Linker1-CD19VL-linker4-CD19VH；
(2)CD19VL-linker4-CD19VH-Linker1-CD22VL-linker4-CD22VH；
(3)CD22VL-linker4-CD22VH-Linker2-CD19VL-linker4-CD19VH；
(4)CD22VL-linker3-CD19VL-linker4-CD19VH-linker3-CD22VH；
(5)CD22VH-linker3-CD19VL-linker4-CD19VH-linker3-CD22VL。
优选地，所述linker4包含如SEQ ID NO:5所示的氨基酸序列或其功能性变体；和/或所述linker1包含如SEQ ID NO:6所示的氨基酸序列或其功能性变体；和/或所述linker2包含如SEQ ID NO:7所示的氨基酸序列或其功能性变体；和/或所述linker3包含如SEQ IDNO:8所示的氨基酸序列或其功能性变体。
具体地，对以上结构(1)-结构(5)的双特异性抗体进行说明：
所述结构(1)中，所述CD19VL分别有CD19VL-1、CD19VL-2两种选择，其氨基酸序列分别如SEQ ID NO:3或SEQ ID NO:68所示；所述CD19VH分别有CD19VH-1、CD19VH-2两种选择，其氨基酸序列分别如SEQ ID NO:4或SEQ ID NO:69所示，即结构(1)的双特异性抗体包含如SEQ ID NO:9或SEQ ID NO:76所示的氨基酸序列或其功能性变体；其中，
所述结构(2)中，其双特异性抗体包含如SEQ ID NO:10所示的氨基酸序列或其功能性变体；
所述结构(3)中，其双特异性抗体包含如SEQ ID NO:11所示的氨基酸序列或其功能性变体；
所述结构(4)中，其双特异性抗体包含如SEQ ID NO:12所示的氨基酸序列或其功能性变体；
所述结构(5)中，所述CD22VL分别有CD22VL-1、CD22VL-2两种选择，其氨基酸序列分别如SEQ ID NO:1或SEQ ID NO:66所示；所述CD22VH分别有CD22VH-1、CD22VH-2两种选择，其氨基酸序列分别如SEQ ID NO:2或SEQ ID NO:67所示，即结构(5)的双特异性抗体包含如SEQ ID NO:13或SEQ ID NO:76所示的氨基酸序列或其功能性变体。
进一步，结构(1)-结构(5)中，编码CD22轻链-1(CD22VL-1)的核苷酸序列如SEQ IDNO:36所示，编码CD22重链-1(CD22VH-1)的核苷酸序列如SEQ ID NO:37所示；编码CD22轻链-2(CD22VL-2)的核苷酸序列如SEQ ID NO:70所示，编码CD22重链-2(CD22VH-2)的核苷酸序列如SEQ ID NO:71所示。
其中，编码CD19轻链-1(CD19VL-1)的核苷酸序列如SEQ ID NO:38所示，编码CD19重链-1(CD19VH-1)的核苷酸序列如SEQ ID NO:39所示；编码CD19轻链-2(CD19VL-2)的核苷酸序列如SEQ ID NO:68所示，编码CD19重链-2(CD19VH-2)的核苷酸序列如SEQ ID NO:69所示。
其中，编码连接CD22轻链(CD22轻链-1或CD22轻链-2)和CD22重链(CD22重链-1或CD22重链-2)的linker4的核苷酸序列均如SEQ ID NO:40所示；编码连接CD19轻链-1和CD19重链-1的linker4的核苷酸序列如SEQ ID NO:41所示；编码连接CD19轻链-2和CD19重链-2的linker4的核苷酸序列如SEQ ID NO:74所示。即说明，连接CD22轻链和CD22重链的linker4与连接CD19轻链和CD19重链的linker4的氨基酸序列相同，编码其氨基酸序列的核苷酸不同。
其中，编码连接CD22单链抗体与CD19单链抗体的linker1的核苷酸序列如SEQ IDNO:42所示；编码连接CD22单链抗体与CD19单链抗体的linker2的核苷酸序列如SEQ ID NO:43所示。
其中，编码与CD19轻链(CD19VL)相连的Linker3的核苷酸如SEQ ID NO:44所示，编码与CD19重链(CD19VH)相连的Linker3的核苷酸如SEQ ID NO:45所示。即说明，与CD19轻链(CD19VL)相连的Linker3与CD19重链(CD19VH)相连的Linker3的氨基酸相同，编码其氨基酸的核苷酸序列不同。
本发明进一步还提供一种靶向CD22和CD19的嵌合抗原受体，嵌合抗原受体中的CD22序列可以与现有的所有公开的CD19 scFV序列一起构建双靶点的嵌合抗原受体，或者也可以跟其他靶点序列一起构建双靶点。
所述嵌合抗原受体包括以下两种结构中的一种：
(1)所述嵌合抗原受体依次连接胞外结构域、铰链区、跨膜区、胞内信号域，所述胞外结构域包含前任一所述的双特异性抗体，结构为以下结构中的一种：
(a)CD22VL-linker4-CD22VH-Linker1-CD19VL-linker4-CD19VH-8h-8TM-BBZ；或
(b)CD19VL-linker4-CD19VH-Linker1-CD22VL-linker4-CD22VH-8h-8TM-BBZ；或
(c)CD22VL-linker4-CD22VH-Linker2-CD19VL-linker4-CD19VH-8h-8TM-BBZ；或
(d)CD22VL-linker3-CD19VL-linker4-CD19VH-linker3-CD22VH-8h-8TM-BBZ；或
(e)CD22VH-linker3-CD19VL-linker4-CD19VH-linker3-CD22VL-8h-8TM-BBZ；或
(2)CD19VL-linker4-CD19VH-8h-8TM-BBZ-2A-CD22VL-linker4-CD22VH-8h`-28TM-28Z`；
所述结构(2)中，CD19VL包含如SEQ ID NO:3所示的氨基酸序列或其功能性变体，CD19VH包含如SEQ ID NO:4所示的氨基酸序列或其功能性变体，CD22VL包含如SEQ ID NO:1所示的氨基酸序列或其功能性变体，CD22VH包含如SEQ ID NO:3所示的氨基酸序列或其功能性变体，linker4包含如SEQ ID NO:5所示的氨基酸序列或其功能性变体。
进一步，结构(2)中，所述2A包含如SEQ ID NO:14、SEQ ID NO:60或SEQ ID NO:61所示的氨基酸序列或其功能性变体，编码其氨基酸序列的核苷酸序列如SEQ ID NO:46、SEQID NO:62或SEQ ID NO:63所示。
优选地，上述嵌合抗原受体的结构中，所述8h包含如SEQ ID NO:15所示的氨基酸序列或其功能性变体；所述8h`包含如SEQ ID NO:16所示的氨基酸序列或其功能性变体。其中，结构(1)的(a)-(e)中，编码铰链区(8h)的氨基酸序列的核苷酸序列如SEQ ID NO:47所示；结构(2)中，编码铰链区(8h)的氨基酸序列的核苷酸序列如SEQ ID NO:48所示；编码铰链区(8h`)的氨基酸序列的核苷酸序列如SEQ ID NO:49所示。即说明，结构(1)中的8h铰链区与结构(2)中的铰链区(8h)的氨基酸序列相同，编码其氨基酸序列的核苷酸不同。
优选地，上述嵌合抗原受体的结构中，所述8TM包含如SEQ ID NO:17所示的氨基酸序列或其功能性变体；所述28TM包含如SEQ ID NO:18所示的氨基酸序列或其功能性变体。其中，结构(1)的(a)-(e)中，编码跨膜区(8TM)的氨基酸序列的核苷酸序列如SEQ ID NO:50所示；结构(2)中，编码跨膜区(8TM)的氨基酸序列的核苷酸序列如SEQ ID NO:51所示；编码跨膜区(28TM)的氨基酸序列的核苷酸序列如SEQ ID NO:52所示。即说明，结构(1)中的跨膜区(8TM)与结构(2)的跨膜区(8TM)的氨基酸序列相同，编码其氨基酸序列的核苷酸不同。
优选地，上述嵌合抗原受体的结构中，所述BBZ中Z的包含如SEQ ID NO:19所示的氨基酸序列或其功能性变体，所述BBZ中的BB包含如SEQ ID NO:20所示的氨基酸序列或其功能性变体；所述28Z`中的Z`包含如SEQ ID NO:21所示的氨基酸序列或其功能性变体，所述28Z`中的28包含如SEQ ID NO:22所示的氨基酸序列或其功能性变体。其中，结构(1)的(a)-(e)中，编码信号传导区(Z)的氨基酸的核苷酸序列如SEQ ID NO:53所示，编码共刺激结构域(BB)的氨基酸的核苷酸序列如SEQ ID NO:54所示；结构(2)中，编码信号传导区(Z)的氨基酸的核苷酸序列如SEQ ID NO:55所示，编码共刺激结构域(BB)的氨基酸的核苷酸序列如SEQ ID NO:56所示；编码信号传导区(Z`)的氨基酸的核苷酸序列如SEQ ID NO:57所示，编码共刺激结构域(28)的氨基酸的核苷酸序列如SEQ ID NO:58所示。即说明结构(1)中的信号传导区(Z)的氨基酸与结构(2)中的信号传导区(Z)的氨基酸相同，编码其氨基酸的核苷酸不同；结构(1)中的共刺激结构域(BB)的氨基酸与结构(2)中的共刺激结构域(BB)的氨基酸相同，编码其氨基酸的核苷酸不同。
优选地，所述嵌合抗原受体包含如SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ ID NO:77、SEQ ID NO:78、SEQ ID NO:83或SEQ ID NO:84所示的氨基酸序列或其功能性变体。其中，SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:77、SEQ ID NO:78为结构(1)中不同结构对应的嵌合抗原受体；SEQ ID NO:29、SEQ ID NO:83、SEQ ID NO:84为结构(2)中，不同的2A对应的嵌合抗原受体。
进一步，所述结构(1)中，嵌合抗原受体连接信号肽，所述信号肽包含如SEQ IDNO:23所示的氨基酸序列或其功能性变体；或所述结构(2)中，由2A连接的两组嵌合抗原受体中至少一组连接信号肽，所述信号肽包含如SEQ ID NO:23所示的氨基酸序列或其功能性变体。其中，编码所述信号肽的核苷酸序列如SEQ ID NO:59或SEQ ID NO:65所示。
其中，连接结构(e)的信号肽，若结构(e)中的CD22VL选自CD22VL-2和CD22VH-2(SEQ ID NO:66、SEQ ID NO:67)时，编码连接结构(e)的信号的核酸序列如SEQ ID NO:65所示。其余结构如SEQ ID NO:59所示。
在某些实施例中，所述CAR结构包含天然杀伤细胞受体(NKR)的一种或多种组分，因而形成NKR-CAR。NKR组分可以是来自以下任何天然杀伤细胞受体的跨膜结构域、铰链结构域或胞质结构域：杀伤细胞免疫球蛋白样受体(KIR)，例如KIR2DL1、KIR2DL2/L3、KIR2DL4、KIR2DL5A、KIR2DL5B、KIR2DS1、KIR2DS2、KIR2DS3、KIR2DS4、DIR2DS5、KIR3DL1/S1、KIR3DL2、KIR3DL3、KIR2DP1和KIR3DP1；天然细胞毒性受体(NCR)，例如，NKp30、NKp44、NKp46；免疫细胞受体的信号传导淋巴细胞活化分子(SLAM)家族，例如，CD48、CD229、2B4、CD84、NTB-A、CRA、BLAME和CD2F-10；Fc受体(FcR)，例如，CD16、和CD64；和Ly49受体，例如，LY49A、LY49C。所述的NKR-CAR分子可以与衔接分子或胞内信号结构域(例如，DAP12)相互作用。
本发明目的在于还提供一种核酸序列，所述核酸序列编码前任一所述的嵌合抗原受体。
具体地：编码上述嵌合抗原受体结构(1)中的(a)的核酸序列如sequence NO.30或SEQ ID NO:82所示；编码上述嵌合抗原受体结构(1)中的(b)的核酸序列如sequence NO.31所示；编码上述嵌合抗原受体结构(1)中的(c)的的核酸序列如sequence NO.32所示；编码上述嵌合抗原受体结构(1)中的(d)的的核酸序列如sequence NO.33所示；编码上述嵌合抗原受体结构(1)中的(e)的的核酸序列如sequence NO.34或SEQ ID NO:82所示；编码上述嵌合抗原受体结构(2)的核酸序列如sequence NO.35、SEQ ID NO:85或SEQ ID NO:86所示。
本发明目的在于还提供一种包含前任一所述的核酸序列的表达载体，比如包含编码前述上述嵌合抗原受体结构(1)中(a)-(e)或结构(2)核酸序列。
进一步，所述核酸序列中还连接启动子，所述启动子选自巨细胞病毒立即早期基因启动子(CMV)、延伸因子1α启动子(EF1-α)、磷酸甘油酸激酶-1启动子(PGK)、泛素-C启动子(UBQ-C)、巨细胞病毒增强子/鸡β-肌动蛋白启动子(CAG)、多瘤病毒增强子/单纯疱疹胸苷激酶启动子(MC1)、β肌动蛋白启动子(β-ACT)、猿猴病毒40启动子(SV40)、骨髓增生肉瘤病毒增强子、阴性对照区缺失的dl587rev引物结合位点取代的(MND)启动子、缺氧启动子中一种。
进一步，所述缺氧启动子的序列如SEQ ID NO:62所示。
进一步，所述表达载体选自慢病毒表达载体、逆转录病毒表达载体、腺病毒表达载体、腺相关病毒表达载体、DNA载体，RNA载体、质粒中的任一种。
在某些实施例中，所述慢病毒载体选自基本上由以下组成的群组：人免疫缺陷病毒1(HIV-1)、人免疫缺陷病毒2(HIV-2)、维斯纳-梅迪病毒(visna-maedi virus，VMV)病毒、山羊关节炎-脑炎病毒(CAEV)、马传染性贫血病毒(EIAV)、猫免疫缺陷病毒(FIV)、牛免疫缺陷病毒(BIV)和猿猴免疫缺陷病毒(SIV)。
在某些实施例中，载体包含左(5')逆转录病毒LTR、Psi(Ψ)包装信号、中心多嘌呤段/DNA瓣(cPPT/FLAP)、逆转录病毒导出元件、可操作地连接到编码本文所涵盖的CAR的多核苷酸的启动子和右(3')逆转录病毒LTR。
在某些实施例中，CAR包含乙型肝炎病毒转录后调节元件(HPRE)或土拔鼠转录后调节元件(WPRE)以及优化的土拔鼠转录后调节元件(oPRE)。
在某些实施例中，所述5'LTR的启动子经异源启动子置换。
在某些实施例中，所述异源启动子是巨细胞病毒(CMV)启动子、劳斯肉瘤病毒(Rous Sarcoma Virus，RSV)启动子或猿猴病毒40(SV40)启动子。
在某些实施例中，所述5'LTR或3'LTR是慢病毒LTR。
在某些实施例中，所述3'LTR是自我失活(SIN)LTR。
在某些实施例中，编码本文所涵盖的CAR的多核苷酸包含优化的Kozark序列。
在某些实施例中，包含CAR的载体可以包含分泌型抗PD-1ScFv；在某些实施例中，包含CAR的载体包含PD-1共轭转导肽(如PD-1-CD28-CD137-CD3信号结构)；在某些实施例中，包含CAR的载体两个以上CAR组合，如2个以上靶向不同抗原或同一抗原的不同识别位点的CAR组合。
本发明目的在于还提供一种工程化的细胞，所述细胞中转导有前任一所述的核酸序列或前任一所述的表达载体；优选的，所述细胞为T细胞、T细胞前体或NK细胞。
本发明目的在于还提供一种细胞制品，所述细胞制品包括前任一所述的工程化的细胞。
进一步，所述细胞制品还包括其他可增强CAR表达活性的活性剂。
在某些具体实施例中，所述活性剂为免疫抑制剂，例如环孢素(cyclosporin)、硫唑嘌呤(azathioprine)、甲氨蝶呤(methotrexate)、霉酚酸酯(mycophenolate)和FK506、抗体或其它免疫清除剂(immunoablativeagents)例如CAMPATH、抗CD3抗体或其它抗体治疗、环磷酰胺(cytoxan)、氟达拉滨(fludarabine)、环孢素(cyclosporin)、FK506、雷帕霉素(rapamycin)、霉酚酸(mycophenolicacid)、类固醇(steroids)、FR901228、细胞因子和辐射。
在某些具体实施例中，增强CAR表达细胞活性的活性剂可以是阻断抑制性分子的活性剂。抑制性分子如PD1可以在一些实施方案中降低CAR表达细胞发动免疫效应子反应的能力。抑制性分子包括PD1、PD-L1、CTLA4、TIM3、LAG3、VISTA、BTLA、TIGIT,LAIR1、CD160、2B4、CEACAM(CEACAM-1、CEACAM-3、CEACAM-5)、LAG3、VISTA、BTLA、TIG、LAIR1、CD160、2B4、CD80、CD86、B7-H3(CD276)、B7-H4(VTCN1)、HVEM(TNFRSF14或CD270)、KIR、A2aR、MHC I类、MHC II类、GAL9、腺苷、TGFR(TGFRβ)和TGFRβ。所述抑制性分子的胞外结构域可以融合到跨膜结构域和胞内信号传导结构域，比如PD1 CAR。
进一步，多种另外的治疗剂可以与本文所述的组合物联合使用。例如，潜在有用的另外的治疗剂包括PD-1抑制剂，例如纳武单抗(nivolumab)派姆单抗(pembrolizumab)派姆单抗、阿巴伏单抗(pidilizumab)和阿特珠单抗(Atezolizumab)。
具体地，适合于与本发明组合使用的另外的治疗剂包括但不限于伊布替尼(ibrutinib)奥法木单抗(ofa tumuma b)利妥昔单抗(rituxima b)贝伐单抗(bevacizumab)曲妥珠单抗(tras tuzuma b)trastuzumabemtansine伊马替尼(imatinib)西妥昔单抗(cetuximab)帕尼单抗(panitumumab)卡妥索单抗(Catumaxomab)、替伊莫单抗(ibritumomab)、奥法木单抗、托西莫单抗(tositumomab)、本妥昔单抗(brentuximab)、阿仑单抗(alemtuzumab)、吉妥珠单抗(gemtuzumab)、厄洛替尼(erlotinib)、吉非替尼(gefitinib)、凡德他尼(vandetanib)、阿法替尼afatinib)、拉帕替尼(lapatinib)、来那替尼(neratinib)、阿昔替尼(axitinib)、马赛替尼(masitini b)、帕唑帕尼(pa zopa ni b)、舒尼替尼(sunitini b)、索拉非尼(sorafenib)、toceranib、来他替尼(lestaurtinib)、阿昔替尼(axitinib)、西地尼布(cediranib)、乐伐替尼(lenvatinib)、尼达尼布(nintedanib)、帕唑帕尼(pazopanib)、瑞格非尼(regorafenib)、司马沙尼(semaxanib)、索拉非尼(sorafenib)、舒尼替尼(sunitinib)、tivozanib、toceranib、凡德他尼、恩曲替尼(entrectinib)、卡博替尼(cabozantinib)、伊马替尼(imatinib)、达沙替尼(dasatinib)、尼洛替尼(nilotinib)、帕纳替尼(pona tini b)、雷多替尼(rad otini b)、博舒替尼(bosutini b)、来他替尼(lestaurtinib)、鲁索替尼(ruxolitinib)、pacritinib、考比替尼(cobimetinib)、司美替尼(selumetinib)、曲美替尼(trametinib)、binimetinib、阿雷替尼(alectinib)、色瑞替尼(ceritinib)、克唑替尼(crizotinib)、阿柏西普(aflibercept)、adipotide、地尼白介素。
进一步，所述细胞制品还可以与一些治疗手段组合使用，所述治疗手段可以是手术、化疗、放射。
本发明目的在于还提供一种药物组合物，所述药物组合物包括前任一所述的双特异性抗体或前任一所述的嵌合抗原受体或前任一所述的核酸序列或前任一所述的表达载体或前任一所述的工程化的细胞或前任一所述的细胞制品。
本发明目的在于还提供一种前任一所述的双特异性抗体或前任一所述的嵌合抗原受体或前任一所述的核酸序列或前任一所述的表达载体或前任一所述的工程化的细胞或前任一所述的细胞制品或前任一所述的药物组合物在制备抗肿瘤药物中的应用。
进一步，所述抗肿瘤药物为抗急性淋巴样白血病药物、抗慢性淋巴细胞白血病药物、抗慢性髓性白血病药物、抗非霍奇金淋巴瘤药物、抗霍奇金淋巴瘤药物、抗前列腺癌药物、抗结直肠癌药物、抗乳腺癌药物、抗卵巢癌药物、抗宫颈癌药物、抗胰腺癌药物、抗肺癌药物、抗肾癌药物、抗肝癌药物、抗脑癌药物或抗皮肤癌药物。
本发明中，“BB”为共刺激结构域CD137的简写，Z为信号传导区CD3ζ的简写，28为共刺激结构域CD28的简写。
本发明中，术语“功能性变体”通常是指包括与其具有基本上相同的功能(例如，可以具备所述嵌合抗原受体的性质)，且与其具有至少85％(例如，至少85％，至少90％，至少91％，至少92％，至少93％，至少94％，至少95％，至少96％，至少97％，至少98％，至少99％，或至少100％)序列同一性的氨基酸序列。在某些实施方式中，所述氨基酸序列的变体为与其具有基本上相同的功能。
本发明中，“h”和“h`”代表铰链区，“TM”代表跨膜区，“BB”代表CD137,“Z”和“Z`”代表CD3ζ。
本发明有益效果在于
本发明提供的靶向CD22和CD19的双特异性抗体及嵌合抗原受体克服了现有技术中双靶点CAR的技术效果缺陷，可以同时靶向两个不同的肿瘤抗原，提高对肿瘤细胞杀伤，减少其免疫逃逸的几率，降低CAR-T治疗后肿瘤复发率，且CAR转导效率高，适合工业化生产，经试验验证可同时靶向CD19和CD22的双特异性位点的CAR-T细胞具有很好的抗肿瘤效果。
附图说明
图1为dCAR1、dCAR4对不同靶细胞的杀伤效果。
图2为dCAR1、dCAR3、dCAR5对不同靶细胞的杀伤效果。
图3为dCAR1、dCAR3、dCAR6对不同靶细胞的杀伤效果。
图4为dCAR2对不同靶细胞的杀伤效果。
图5为静脉注射dCAR1、dCAR2、dCAR4后小鼠体内活体成像的荧光曲线变化。
图6为静脉注射dCAR1、dCAR6后小鼠体内活体成像的荧光曲线变化。
图7为dCAR8、dCAR9对不同靶细胞的杀伤效果。
图8为dCAR1、dCAR7对不同靶细胞的杀伤效果。
具体实施方式
所举实施例是为了更好地对本发明进行说明，但并不是本发明的内容仅局限于所举实施例。所以熟悉本领域的技术人员根据上述发明内容对实施方案进行非本质的改进和调整，仍属于本发明的保护范围。
实施例1靶向CD19和CD22的CAR T质粒构建
通过PCR扩增获得CD19或CD22 ScFv，再通过限制性内酶酶切的方式，按不同排列顺序组合，将序列连接于含不同Linker，相同信号结构的慢病毒载体中，即获得不同结构的靶向CD19和CD22的双靶向CAR T质粒，结构如下：
dCAR1：CD22VL-linker4-CD22VH-Linker1-CD19(1)VL-linker4-CD19(1)VH-8h(a)-8TM-BBZ(a)；
dCAR2：CD19(1)VL-linker4-CD19(1)VH-Linker1-CD22VL-linker4-CD22VH-8h(a)-8TM-BBZ(a)；
dCAR3：CD22VL-linker4-CD22VH-Linker2-CD19(1)VL-linker4-CD19(1)VH-8h(a)-8TM-BBZ(a)；
dCAR4：CD22VL-linker3-CD19(1)VL-linker4-CD19(1)VH-linker3-CD22VH-8h(a)-8TM-BBZ(a)；
dCAR5：CD22VH-linker3-CD19(1)VL-linker4-CD19(1)VH-linker3-CD22VL-8h(a)-8TM-BBZ(a)；
dCAR6：CD19VL-linker4-CD19VH-8h(a)-8TM-BBZ(a)-2A-CD8a-CD22VL-linker4-CD22VH-8h(b)-28TM-28Z(b)；
dCAR7：CD22VH(m971)-linker3-CD19VL-linker4-CD19VH-linker3-CD22VH(m971)-8h(a)-8TM-BBZ(a)；
dCAR8:CD8a-CD22VL-linker4-CD22VH-Linker1-CD19(2)VL-linker4-CD19(2)VH-8h(a)-8TM-BBZ(a)；
dCAR9：启动子-CD8a-CD22VL-linker4-CD22VH-Linker1-CD19(2)VL-linker4-CD19(2)VH-8h(a)-8TM-BBZ(a)；
其中，以上结构中，各个部件的氨基酸序列和核苷酸序列如下表1所示,
表1 CAR结构中各个部件的氨基酸和核苷酸序列表
Figure BDA0003394632760000111
实施例2制备慢病毒及感染T淋巴细胞
本实施例包装慢病毒采用磷酸钙法，具体为：用含10％FBS(w/v)的DMEM培养基培养293T细胞至较佳状态，包装质粒(RRE:REV:2G)和表达质粒按一定比列加入到1.5的离心管中，加入CaCl2和2×HBS，混匀后室温静置后加入到处理好的293T细胞培养液中，3-5h后再次换液至10mL含10％FBS的DMEM培养基，48h或72h后收集细胞上清，纯化病毒，进行滴度测定。
制备的慢病毒感染CHO细胞，分别将感染CHO细胞后的CD19 CAR-T、CD22 CAR-T、双CAR-T(19和22)采用CD19-FC，CD22-His流式标记试剂标记后检测双CAR结构中靶向CD19CAR和靶向CD22 CAR的表达，用Protein-L检测总的CAR表达，结果如图1所示：CD19 CAR-T仅能识别CD19-FC，CD22 CART仅能识别CD22-His，而双CAR-T(19和22)可以识别CD19-FC和CD22-His双靶点。
利用梯度离心法进行淋巴细胞分离；离心后，取第二层白色淋巴细胞层，生理盐水洗涤，加入含有10％FBS的RPMI 1640完全培养基培养，获得人PBMC细胞。获得的PBMC细胞经抗CD3、CD28单克隆抗体活化24h后，按一定的感染复数(MOI)感染已活化的PBMC，在病毒感染的第12天检测CAR-T的阳性率，检测方法为流式检测，抗体为：Protein-L-PE，Protein-L可识别抗体轻链，CAR抗原识别区的ScFv序列的轻链可被Protein-L识别，因此利用Protein-L可检测CAR阳性率和CAR表达强度。
实施例3
以CD19+CD22-Nalm6、CD19-CD22+Nalm6和Nalm6细胞作为阳性靶细胞，将CAR T细胞按1:2的比例铺于靶细胞中，并利用ACEA xCELLigence RTCA MP仪器检测不同CAR-T对靶细胞杀伤能力，实验步骤依据仪器说明书进行。
结果如表1和图1所示，加入CAR-T细胞24小时后，dCAR1、dCAR4对CD19+CD22-Nalm6、CD19-CD22+Nalm6和Nalm6靶细胞的杀伤效果均显著高于Control T组。
表1 dCAR1、dCAR4对不同靶细胞的杀伤效果
Figure BDA0003394632760000121
实施例4
以CD19+CD22-Nalm6、CD19-CD22+Nalm6和Nalm6细胞作为阳性靶细胞，将CAR T细胞按1:1的比例铺于靶细胞中，并利用ACEA xCELLigence RTCA MP仪器检测不同CAR-T对靶细胞杀伤能力，实验步骤依据仪器说明书进行。
结果如表2、表3和图2、图3所示，加入CAR-T细胞24小时后，dCAR1、dCAR3、dCAR5和dCAR6对CD19+CD22-Nalm6、CD19-CD22+Nalm6和Nalm6靶细胞均有明显的杀伤效果。
表2 dCAR1、dCAR3、dCAR5对不同靶细胞的杀伤效果
Figure BDA0003394632760000131
表3 dCAR1、dCAR3、dCAR6对不同靶细胞的杀伤效果
Figure BDA0003394632760000132
实施例5
以CD19-CD22+Nalm6和Nalm6细胞作为阳性靶细胞，CD19-CD22-Nalm6细胞作为阴性靶细胞，将CAR T细胞按1:4的比例铺于靶细胞中，并利用ACEA xCELLigence RTCA MP仪器检测不同CAR-T对靶细胞杀伤能力，实验步骤依据仪器说明书进行。
结果如表4和图4所示：加入CAR-T细胞24小时后，dCAR2对CD19-CD22+Nalm6、Nalm6和CD19-CD22-Nalm6靶细胞均有杀伤效果。
表4 dCAR2对不同靶细胞的杀伤效果
Figure BDA0003394632760000133
实施例6
选择6-8周鼠龄雌性NCG小鼠，在小鼠尾部以1×10^6/只细胞量静脉注射Nalm6-Luc-GFP细胞，构建人CD19+CD22+血液瘤荷瘤模型。在成瘤21天，尾静脉注射1*10^6CAR-T细胞/只；Control T组于第21天回输总数相同的T淋巴细胞。活体成像的荧光曲线结果如图5所示，相比于Control T对照组，dCAR1具有明显的体内药效。
实施例7
选择6-8周鼠龄雌性NOG小鼠，在小鼠尾部以每只1×10^6的患者肿瘤细胞进行静脉注射，构建人CD19+CD22+血液瘤荷瘤模型。在成瘤7天，尾静脉注射1*10^6CAR-T细胞/只；Control T组于第7天回输总数相同的T淋巴细胞。活体成像的荧光曲线结果如图6所示，相比于Control T对照组，dCAR1、dCAR6均具有明显的体内药效。
实施例8
以CD19+CD22-Nalm6、CD19-CD22+Nalm6和Nalm6细胞作为阳性靶细胞，将CAR T细胞按1:1的比例铺于靶细胞中，并利用ACEA xCELLigence RTCA MP仪器检测不同CAR-T对靶细胞杀伤能力，实验步骤依据仪器说明书进行。
结果如图7所示，与control T相比，dCAR8与dCAR9对CD22阳性的靶细胞具有明显的杀伤能力。
实施例9
以CD19+CD22-Nalm6、CD19-CD22+Nalm6和Nalm6细胞作为阳性靶细胞，将CAR T细胞按1:2的比例铺于靶细胞中，并利用ACEA xCELLigence RTCA MP仪器检测不同CAR-T对靶细胞杀伤能力，实验步骤依据仪器说明书进行。
结果如图8所示，可以看出现有的采用m971制备的双CAR对CD22阳性的靶细胞无杀伤能力，而采用本发明中的CD22抗体制备的双CAR(dCA1、dCAR7)对CD22阳性的靶细胞具有明显的杀伤能力。
最后说明的是，以上实施例仅用以说明本发明的技术方案而非限制，尽管参照较佳实施例对本发明进行了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的宗旨和范围，其均应涵盖在本发明的权利要求范围。
序列表
<110> 重庆精准生物技术有限公司，重庆精准生物产业技术研究院有限公司
<120> 靶向CD22和CD19的双特异性抗体及其嵌合抗原受体和应用
<130> 2021-11-30
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500 505
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Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met
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Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
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Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe
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Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu
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Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu
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Pro Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr
210 215 220
Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln
225 230 235 240
Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr
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Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly
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Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly
275 280 285
Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr
290 295 300
Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr
305 310 315 320
Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys
325 330 335
His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly
340 345 350
Ser Ser Val Thr Val Ser Ser Glu Ala Ala Ala Lys Glu Ala Ala Ala
355 360 365
Lys Glu Ala Ala Ala Lys Gly Gly Gly Gly Ser Gln Val Gln Leu Val
370 375 380
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser
385 390 395 400
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly Ile Ser Trp Val
405 410 415
Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Trp Ile Ser Ala
420 425 430
Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln Gly Arg Val Thr
435 440 445
Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met Glu Leu Arg Ser
450 455 460
Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Phe Gln
465 470 475 480
Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val
485 490 495
Thr Val Ser Ser
500
<210> 13
<211> 485
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu
50 55 60
Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile
115 120 125
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg
130 135 140
Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn
145 150 155 160
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile Tyr His
165 170 175
Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly
180 185 190
Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp
195 200 205
Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr Thr Phe
210 215 220
Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly Ser Gly
225 230 235 240
Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Gln
245 250 255
Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr
260 265 270
Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile
275 280 285
Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile Trp Gly
290 295 300
Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu Thr Ile
305 310 315 320
Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr Asn Met
325 330 335
Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr
340 345 350
Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser Val Thr
355 360 365
Val Ser Ser Gly Gly Gly Gly Ser Glu Thr Thr Leu Thr Gln Ser Pro
370 375 380
Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg
385 390 395 400
Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro
405 410 415
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr
420 425 430
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
435 440 445
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Tyr Cys
450 455 460
Gln Gln Tyr His Ser Trp Pro Pro Leu Thr Phe Gly Gly Gly Thr Lys
465 470 475 480
Leu Glu Ile Lys Arg
485
<210> 14
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val
1 5 10 15
Glu Glu Asn Pro Gly Pro
20
<210> 15
<211> 45
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala
1 5 10 15
Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly
20 25 30
Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp
35 40 45
<210> 16
<211> 24
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu
1 5 10 15
Ser Leu Val Ile Thr Leu Tyr Cys
20
<210> 17
<211> 27
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu
1 5 10 15
Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val
20 25
<210> 18
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly
1 5 10 15
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr
20 25 30
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys
35 40 45
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys
50 55 60
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg
65 70 75 80
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala
85 90 95
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
100 105 110
<210> 19
<211> 42
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met
1 5 10 15
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe
20 25 30
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu
35 40
<210> 20
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly
1 5 10 15
Gln Ser Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr
20 25 30
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys
35 40 45
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys
50 55 60
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg
65 70 75 80
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala
85 90 95
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
100 105 110
<210> 21
<211> 41
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Arg Ser Lys Arg Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr
1 5 10 15
Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro
20 25 30
Pro Arg Asp Phe Ala Ala Tyr Arg Ser
35 40
<210> 22
<211> 21
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro
20
<210> 23
<211> 731
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Tyr His Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Thr
100 105 110
Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln
115 120 125
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser
130 135 140
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly
145 150 155 160
Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
165 170 175
Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln
180 185 190
Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met
195 200 205
Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln
225 230 235 240
Gly Thr Leu Val Thr Val Ser Ser Glu Ala Ala Ala Lys Glu Ala Ala
245 250 255
Ala Lys Glu Ala Ala Ala Lys Asp Ile Gln Met Thr Gln Ser Pro Ser
260 265 270
Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala
275 280 285
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly
290 295 300
Lys Ala Pro Arg Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
305 310 315 320
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu
325 330 335
Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
340 345 350
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu
355 360 365
Ile Lys Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly
370 375 380
Ser Thr Lys Gly Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val
385 390 395 400
Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser
405 410 415
Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala
420 425 430
Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser
435 440 445
Thr Ser Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn
450 455 460
Gln Val Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr
465 470 475 480
Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp
485 490 495
Tyr Trp Gly Gln Gly Ser Ser Val Thr Val Ser Ser Thr Thr Thr Pro
500 505 510
Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu
515 520 525
Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His
530 535 540
Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu
545 550 555 560
Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr
565 570 575
Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe
580 585 590
Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg
595 600 605
Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser
610 615 620
Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln Leu Tyr
625 630 635 640
Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys
645 650 655
Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn
660 665 670
Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu
675 680 685
Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly
690 695 700
His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr
705 710 715 720
Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
725 730
<210> 24
<211> 731
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
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 Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln
115 120 125
Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser
130 135 140
Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser
145 150 155 160
Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile
165 170 175
Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu
180 185 190
Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr
195 200 205
Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr
210 215 220
Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser
225 230 235 240
Val Thr Val Ser Ser Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu
245 250 255
Ala Ala Ala Lys Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr Leu Ser
260 265 270
Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
275 280 285
Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
290 295 300
Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala
305 310 315 320
Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser
325 330 335
Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Tyr His
340 345 350
Ser Trp Pro Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
355 360 365
Arg Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser
370 375 380
Thr Lys Gly Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
385 390 395 400
Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe
405 410 415
Thr Ser Tyr Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
420 425 430
Glu Trp Met Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala
435 440 445
Gln Lys Leu Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser
450 455 460
Thr Ala Tyr Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val
465 470 475 480
Tyr Tyr Cys Ala Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp
485 490 495
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Thr Thr Thr Pro
500 505 510
Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu
515 520 525
Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His
530 535 540
Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu
545 550 555 560
Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr
565 570 575
Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe
580 585 590
Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg
595 600 605
Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser
610 615 620
Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln Leu Tyr
625 630 635 640
Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys
645 650 655
Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn
660 665 670
Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu
675 680 685
Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly
690 695 700
His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr
705 710 715 720
Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
725 730
<210> 25
<211> 751
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Tyr His Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Thr
100 105 110
Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln
115 120 125
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser
130 135 140
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly
145 150 155 160
Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
165 170 175
Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln
180 185 190
Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met
195 200 205
Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln
225 230 235 240
Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
245 250 255
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met
260 265 270
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr
275 280 285
Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr
290 295 300
Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile Tyr His Thr Ser
305 310 315 320
Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly
325 330 335
Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala
340 345 350
Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly
355 360 365
Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly Ser Gly Lys Pro
370 375 380
Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Gln Glu Ser
385 390 395 400
Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr
405 410 415
Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln
420 425 430
Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu
435 440 445
Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu Thr Ile Ser Lys
450 455 460
Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr Asn Met Asp Pro
465 470 475 480
Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly
485 490 495
Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser Val Thr Val Ser
500 505 510
Ser Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys
515 520 525
Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala
530 535 540
Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly
545 550 555 560
Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile
565 570 575
Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val
580 585 590
Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe
595 600 605
Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly
610 615 620
Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg
625 630 635 640
Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln
645 650 655
Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp
660 665 670
Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro
675 680 685
Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp
690 695 700
Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg
705 710 715 720
Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr
725 730 735
Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
740 745 750
<210> 26
<211> 723
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Tyr His Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Gly Gly
100 105 110
Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser
115 120 125
Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser
130 135 140
Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu
145 150 155 160
Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe
165 170 175
Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu
180 185 190
Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu
195 200 205
Pro Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr
210 215 220
Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln
225 230 235 240
Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr
245 250 255
Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly
260 265 270
Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly
275 280 285
Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr
290 295 300
Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr
305 310 315 320
Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys
325 330 335
His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly
340 345 350
Ser Ser Val Thr Val Ser Ser Glu Ala Ala Ala Lys Glu Ala Ala Ala
355 360 365
Lys Glu Ala Ala Ala Lys Gly Gly Gly Gly Ser Gln Val Gln Leu Val
370 375 380
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser
385 390 395 400
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly Ile Ser Trp Val
405 410 415
Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Trp Ile Ser Ala
420 425 430
Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln Gly Arg Val Thr
435 440 445
Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met Glu Leu Arg Ser
450 455 460
Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Phe Gln
465 470 475 480
Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val
485 490 495
Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala
500 505 510
Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg
515 520 525
Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys
530 535 540
Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu
545 550 555 560
Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu
565 570 575
Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln
580 585 590
Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly
595 600 605
Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr
610 615 620
Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg
625 630 635 640
Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met
645 650 655
Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu
660 665 670
Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys
675 680 685
Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu
690 695 700
Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu
705 710 715 720
Pro Pro Arg
<210> 27
<211> 708
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu
50 55 60
Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile
115 120 125
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg
130 135 140
Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn
145 150 155 160
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile Tyr His
165 170 175
Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly
180 185 190
Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp
195 200 205
Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr Thr Phe
210 215 220
Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly Ser Gly
225 230 235 240
Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Gln
245 250 255
Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr
260 265 270
Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile
275 280 285
Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile Trp Gly
290 295 300
Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu Thr Ile
305 310 315 320
Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr Asn Met
325 330 335
Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr
340 345 350
Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser Val Thr
355 360 365
Val Ser Ser Gly Gly Gly Gly Ser Glu Thr Thr Leu Thr Gln Ser Pro
370 375 380
Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg
385 390 395 400
Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro
405 410 415
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr
420 425 430
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
435 440 445
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Tyr Cys
450 455 460
Gln Gln Tyr His Ser Trp Pro Pro Leu Thr Phe Gly Gly Gly Thr Lys
465 470 475 480
Leu Glu Ile Lys Arg Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
485 490 495
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
500 505 510
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
515 520 525
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
530 535 540
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys
545 550 555 560
Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr
565 570 575
Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly
580 585 590
Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala
595 600 605
Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg
610 615 620
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu
625 630 635 640
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn
645 650 655
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met
660 665 670
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly
675 680 685
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala
690 695 700
Leu Pro Pro Arg
705
<210> 28
<211> 965
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
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 Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln
115 120 125
Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser
130 135 140
Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser
145 150 155 160
Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile
165 170 175
Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu
180 185 190
Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr
195 200 205
Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr
210 215 220
Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser
225 230 235 240
Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
245 250 255
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
260 265 270
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
275 280 285
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
290 295 300
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys
305 310 315 320
Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr
325 330 335
Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly
340 345 350
Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala
355 360 365
Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg
370 375 380
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu
385 390 395 400
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn
405 410 415
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met
420 425 430
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly
435 440 445
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala
450 455 460
Leu Pro Pro Arg Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln
465 470 475 480
Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Glu Thr Thr Leu Thr Gln
485 490 495
Ser Pro Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser
500 505 510
Cys Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln
515 520 525
Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg
530 535 540
Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu
545 550 555 560
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr
565 570 575
Tyr Cys Gln Gln Tyr His Ser Trp Pro Pro Leu Thr Phe Gly Gly Gly
580 585 590
Thr Lys Leu Glu Ile Lys Arg Gly Ser Thr Ser Gly Ser Gly Lys Pro
595 600 605
Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Val Gln Ser
610 615 620
Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys
625 630 635 640
Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly Ile Ser Trp Val Arg Gln
645 650 655
Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Trp Ile Ser Ala Tyr Asn
660 665 670
Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln Gly Arg Val Thr Met Thr
675 680 685
Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met Glu Leu Arg Ser Leu Arg
690 695 700
Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Phe Gln Gly Ile
705 710 715 720
Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
725 730 735
Ser Ser Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala
740 745 750
Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg
755 760 765
Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys
770 775 780
Asp Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser
785 790 795 800
Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg
805 810 815
Ser Arg Gly Gly His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro
820 825 830
Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe
835 840 845
Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro
850 855 860
Ala Tyr Gln Gln Gly Gln Ser Gln Leu Tyr Asn Glu Leu Asn Leu Gly
865 870 875 880
Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro
885 890 895
Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr
900 905 910
Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly
915 920 925
Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln
930 935 940
Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln
945 950 955 960
Ala Leu Pro Pro Arg
965
<210> 29
<211> 2193
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 240
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 300
ggagggacca agctggagat caaacgtgga agcaccagcg gcagcggaaa gcctggcagc 360
ggcgagggca gcacaaaagg acaggtccag ctggtacagt ctggagctga ggtgaagaag 420
cctggggcct cagtgaaggt ctcctgcaag gcttctggtt acacctttac cagctatggt 480
atcagctggg tgcgacaggc ccctggacaa gggcttgagt ggatgggatg gatcagcgct 540
tacaatggta acacaaacta tgcacagaag ctccagggca gagtcaccat gaccacagac 600
acatccacga gcacagccta catggagctg aggagcctga gatctgacga cacggccgtg 660
tattactgtg cgagagactt ccagggtata gcagtggctg atcttgacta ctggggccag 720
ggaaccctgg tcacagtctc ctcagaggcc gctgcaaagg aagcagctgc caaagaggct 780
gcagccaagg acatccagat gacacagtcc cccagctccc tgtctgccag cgtgggcgac 840
cgggtgacca tcacatgcag agcctctcag gatatcagca agtatctgaa ctggtaccag 900
cagaagccag gcaaggcccc caggctgctg atctatcaca cctcccgcct gcactctgga 960
gtgccaagcc ggttctccgg atctggaagc ggaaccgact acaccctgac aatctctagc 1020
ctgcagcctg aggatttcgc cacatactat tgccagcagg gcaataccct gccatataca 1080
tttggcggag gaaccaggct ggagatcaag ggatccacat ctggaagcgg caagccagga 1140
tccggagagg gatctaccaa gggacaggtg cagctgcagg agtccggacc tggactggtg 1200
aagccaagcc agacactgtc cctgacctgt acagtgagcg gcgtgtccct gcctgattac 1260
ggcgtgtcct ggatcagaca gccacctggc aaggccctgg agtggctggg cgtgatctgg 1320
ggctctgaga ccacatacta ttccacctct ctgaagacca ggctgacaat ctctaaggac 1380
aacagcaaga atcaggtggt gctgaccatg acaaacatgg accctgtgga taccgccaca 1440
tactattgtg ccaagcacta ctattacggc ggcagctatg ccatggatta ctggggccag 1500
ggctcctctg tgaccgtgag ctccaccacg acgccagcgc cgcgaccacc aacaccggcg 1560
cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 1620
ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1680
gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg caaacggggc 1740
agaaagaaac tcctgtatat attcaaacaa ccatttatga gaccagtaca aactactcaa 1800
gaggaagatg gctgtagctg ccgatttcca gaagaagaag aaggaggatg tgaactgaga 1860
gtgaagttca gcaggagcgc agacgccccc gcgtacaagc agggccagaa ccagctctat 1920
aacgagctca atctaggacg aagagaggag tacgatgttt tggacaagag acgtggccgg 1980
gaccctgaga tggggggaaa gccgagaagg aagaaccctc aggaaggcct gtacaatgaa 2040
ctgcagaaag ataagatggc ggaggcctac agtgagattg ggatgaaagg cgagcgccgg 2100
aggggcaagg ggcacgatgg cctttaccag ggtctcagta cagccaccaa ggacacctac 2160
gacgcccttc acatgcaggc cctgccccct cgc 2193
<210> 30
<211> 2193
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
gacatccaga tgacacagtc ccccagctcc ctgtctgcca gcgtgggcga ccgggtgacc 60
atcacatgca gagcctctca ggatatcagc aagtatctga actggtacca gcagaagcca 120
ggcaaggccc ccaggctgct gatctatcac acctcccgcc tgcactctgg agtgccaagc 180
cggttctccg gatctggaag cggaaccgac tacaccctga caatctctag cctgcagcct 240
gaggatttcg ccacatacta ttgccagcag ggcaataccc tgccatatac atttggcgga 300
ggaaccaggc tggagatcaa gggatccaca tctggaagcg gcaagccagg atccggagag 360
ggatctacca agggacaggt gcagctgcag gagtccggac ctggactggt gaagccaagc 420
cagacactgt ccctgacctg tacagtgagc ggcgtgtccc tgcctgatta cggcgtgtcc 480
tggatcagac agccacctgg caaggccctg gagtggctgg gcgtgatctg gggctctgag 540
accacatact attccacctc tctgaagacc aggctgacaa tctctaagga caacagcaag 600
aatcaggtgg tgctgaccat gacaaacatg gaccctgtgg ataccgccac atactattgt 660
gccaagcact actattacgg cggcagctat gccatggatt actggggcca gggctcctct 720
gtgaccgtga gctccgaggc cgctgcaaag gaagcagctg ccaaagaggc tgcagccaag 780
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 840
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 900
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 960
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 1020
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 1080
ggagggacca agctggagat caaacgtgga agcaccagcg gcagcggaaa gcctggcagc 1140
ggcgagggca gcacaaaagg acaggtccag ctggtacagt ctggagctga ggtgaagaag 1200
cctggggcct cagtgaaggt ctcctgcaag gcttctggtt acacctttac cagctatggt 1260
atcagctggg tgcgacaggc ccctggacaa gggcttgagt ggatgggatg gatcagcgct 1320
tacaatggta acacaaacta tgcacagaag ctccagggca gagtcaccat gaccacagac 1380
acatccacga gcacagccta catggagctg aggagcctga gatctgacga cacggccgtg 1440
tattactgtg cgagagactt ccagggtata gcagtggctg atcttgacta ctggggccag 1500
ggaaccctgg tcacagtctc ctcaaccacg acgccagcgc cgcgaccacc aacaccggcg 1560
cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 1620
ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1680
gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg caaacggggc 1740
agaaagaaac tcctgtatat attcaaacaa ccatttatga gaccagtaca aactactcaa 1800
gaggaagatg gctgtagctg ccgatttcca gaagaagaag aaggaggatg tgaactgaga 1860
gtgaagttca gcaggagcgc agacgccccc gcgtacaagc agggccagaa ccagctctat 1920
aacgagctca atctaggacg aagagaggag tacgatgttt tggacaagag acgtggccgg 1980
gaccctgaga tggggggaaa gccgagaagg aagaaccctc aggaaggcct gtacaatgaa 2040
ctgcagaaag ataagatggc ggaggcctac agtgagattg ggatgaaagg cgagcgccgg 2100
aggggcaagg ggcacgatgg cctttaccag ggtctcagta cagccaccaa ggacacctac 2160
gacgcccttc acatgcaggc cctgccccct cgc 2193
<210> 31
<211> 2208
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 240
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 300
ggagggacca agctggagat caaacgtgga agcaccagcg gcagcggaaa gcctggcagc 360
ggcgagggca gcacaaaagg acaggtccag ctggtacagt ctggagctga ggtgaagaag 420
cctggggcct cagtgaaggt ctcctgcaag gcttctggtt acacctttac cagctatggt 480
atcagctggg tgcgacaggc ccctggacaa gggcttgagt ggatgggatg gatcagcgct 540
tacaatggta acacaaacta tgcacagaag ctccagggca gagtcaccat gaccacagac 600
acatccacga gcacagccta catggagctg aggagcctga gatctgacga cacggccgtg 660
tattactgtg cgagagactt ccagggtata gcagtggctg atcttgacta ctggggccag 720
ggaaccctgg tcacagtctc ctcaggaggc ggcggttcag gtggtggcgg atctggcgga 780
ggtggttccg gaggtggagg ttcagacatc cagatgacac agtcccccag ctccctgtct 840
gccagcgtgg gcgaccgggt gaccatcaca tgcagagcct ctcaggatat cagcaagtat 900
ctgaactggt accagcagaa gccaggcaag gcccccaggc tgctgatcta tcacacctcc 960
cgcctgcact ctggagtgcc aagccggttc tccggatctg gaagcggaac cgactacacc 1020
ctgacaatct ctagcctgca gcctgaggat ttcgccacat actattgcca gcagggcaat 1080
accctgccat atacatttgg cggaggaacc aggctggaga tcaagggatc cacatctgga 1140
agcggcaagc caggatccgg agagggatct accaagggac aggtgcagct gcaggagtcc 1200
ggacctggac tggtgaagcc aagccagaca ctgtccctga cctgtacagt gagcggcgtg 1260
tccctgcctg attacggcgt gtcctggatc agacagccac ctggcaaggc cctggagtgg 1320
ctgggcgtga tctggggctc tgagaccaca tactattcca cctctctgaa gaccaggctg 1380
acaatctcta aggacaacag caagaatcag gtggtgctga ccatgacaaa catggaccct 1440
gtggataccg ccacatacta ttgtgccaag cactactatt acggcggcag ctatgccatg 1500
gattactggg gccagggctc ctctgtgacc gtgagctcca ccacgacgcc agcgccgcga 1560
ccaccaacac cggcgcccac catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc 1620
cggccagcgg cggggggcgc agtgcacacg agggggctgg acttcgcctg tgatatctac 1680
atctgggcgc ccttggccgg gacttgtggg gtccttctcc tgtcactggt tatcaccctt 1740
tactgcaaac ggggcagaaa gaaactcctg tatatattca aacaaccatt tatgagacca 1800
gtacaaacta ctcaagagga agatggctgt agctgccgat ttccagaaga agaagaagga 1860
ggatgtgaac tgagagtgaa gttcagcagg agcgcagacg cccccgcgta caagcagggc 1920
cagaaccagc tctataacga gctcaatcta ggacgaagag aggagtacga tgttttggac 1980
aagagacgtg gccgggaccc tgagatgggg ggaaagccga gaaggaagaa ccctcaggaa 2040
ggcctgtaca atgaactgca gaaagataag atggcggagg cctacagtga gattgggatg 2100
aaaggcgagc gccggagggg caaggggcac gatggccttt accagggtct cagtacagcc 2160
accaaggaca cctacgacgc ccttcacatg caggccctgc cccctcgc 2208
<210> 32
<211> 2124
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 240
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 300
ggagggacca agctggagat caaacgtggg ggtggaggct ctgacatcca gatgacacag 360
tcccccagct ccctgtctgc cagcgtgggc gaccgggtga ccatcacatg cagagcctct 420
caggatatca gcaagtatct gaactggtac cagcagaagc caggcaaggc ccccaggctg 480
ctgatctatc acacctcccg cctgcactct ggagtgccaa gccggttctc cggatctgga 540
agcggaaccg actacaccct gacaatctct agcctgcagc ctgaggattt cgccacatac 600
tattgccagc agggcaatac cctgccatat acatttggcg gaggaaccag gctggagatc 660
aagggatcca catctggaag cggcaagcca ggatccggag agggatctac caagggacag 720
gtgcagctgc aggagtccgg acctggactg gtgaagccaa gccagacact gtccctgacc 780
tgtacagtga gcggcgtgtc cctgcctgat tacggcgtgt cctggatcag acagccacct 840
ggcaaggccc tggagtggct gggcgtgatc tggggctctg agaccacata ctattccacc 900
tctctgaaga ccaggctgac aatctctaag gacaacagca agaatcaggt ggtgctgacc 960
atgacaaaca tggaccctgt ggataccgcc acatactatt gtgccaagca ctactattac 1020
ggcggcagct atgccatgga ttactggggc cagggctcct ctgtgaccgt gagctccggt 1080
ggaggcgggt ctcaggtcca gctggtacag tctggagctg aggtgaagaa gcctggggcc 1140
tcagtgaagg tctcctgcaa ggcttctggt tacaccttta ccagctatgg tatcagctgg 1200
gtgcgacagg cccctggaca agggcttgag tggatgggat ggatcagcgc ttacaatggt 1260
aacacaaact atgcacagaa gctccagggc agagtcacca tgaccacaga cacatccacg 1320
agcacagcct acatggagct gaggagcctg agatctgacg acacggccgt gtattactgt 1380
gcgagagact tccagggtat agcagtggct gatcttgact actggggcca gggaaccctg 1440
gtcacagtct cctcaaccac gacgccagcg ccgcgaccac caacaccggc gcccaccatc 1500
gcgtcgcagc ccctgtccct gcgcccagag gcgtgccggc cagcggcggg gggcgcagtg 1560
cacacgaggg ggctggactt cgcctgtgat atctacatct gggcgccctt ggccgggact 1620
tgtggggtcc ttctcctgtc actggttatc accctttact gcaaacgggg cagaaagaaa 1680
ctcctgtata tattcaaaca accatttatg agaccagtac aaactactca agaggaagat 1740
ggctgtagct gccgatttcc agaagaagaa gaaggaggat gtgaactgag agtgaagttc 1800
agcaggagcg cagacgcccc cgcgtacaag cagggccaga accagctcta taacgagctc 1860
aatctaggac gaagagagga gtacgatgtt ttggacaaga gacgtggccg ggaccctgag 1920
atggggggaa agccgagaag gaagaaccct caggaaggcc tgtacaatga actgcagaaa 1980
gataagatgg cggaggccta cagtgagatt gggatgaaag gcgagcgccg gaggggcaag 2040
gggcacgatg gcctttacca gggtctcagt acagccacca aggacaccta cgacgccctt 2100
cacatgcagg ccctgccccc tcgc 2124
<210> 33
<211> 2124
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
caggtccagc tggtacagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60
tcctgcaagg cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180
gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240
atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagacttc 300
cagggtatag cagtggctga tcttgactac tggggccagg gaaccctggt cacagtctcc 360
tcagggggtg gaggctctga catccagatg acacagtccc ccagctccct gtctgccagc 420
gtgggcgacc gggtgaccat cacatgcaga gcctctcagg atatcagcaa gtatctgaac 480
tggtaccagc agaagccagg caaggccccc aggctgctga tctatcacac ctcccgcctg 540
cactctggag tgccaagccg gttctccgga tctggaagcg gaaccgacta caccctgaca 600
atctctagcc tgcagcctga ggatttcgcc acatactatt gccagcaggg caataccctg 660
ccatatacat ttggcggagg aaccaggctg gagatcaagg gatccacatc tggaagcggc 720
aagccaggat ccggagaggg atctaccaag ggacaggtgc agctgcagga gtccggacct 780
ggactggtga agccaagcca gacactgtcc ctgacctgta cagtgagcgg cgtgtccctg 840
cctgattacg gcgtgtcctg gatcagacag ccacctggca aggccctgga gtggctgggc 900
gtgatctggg gctctgagac cacatactat tccacctctc tgaagaccag gctgacaatc 960
tctaaggaca acagcaagaa tcaggtggtg ctgaccatga caaacatgga ccctgtggat 1020
accgccacat actattgtgc caagcactac tattacggcg gcagctatgc catggattac 1080
tggggccagg gctcctctgt gaccgtgagc tccggtggag gcgggtctga aacgacactc 1140
acgcagtctc cagccaccct gtctgtgtct ccaggggaaa gagccaccct ctcctgcagg 1200
gccagtcaga gtgttagcag caacttagcc tggtaccagc agaaacctgg ccaggctccc 1260
agactcctca tctatggtgc atccaccagg gccactggca tcccagccag gttcagtggc 1320
agtgggtctg ggaccgagtt cactctcacc atcagcagcc tgcagtctga ggattttgca 1380
atttattact gtcagcagta tcatagctgg cctcccctca ctttcggcgg agggaccaag 1440
ctggagatca aacgtaccac gacgccagcg ccgcgaccac caacaccggc gcccaccatc 1500
gcgtcgcagc ccctgtccct gcgcccagag gcgtgccggc cagcggcggg gggcgcagtg 1560
cacacgaggg ggctggactt cgcctgtgat atctacatct gggcgccctt ggccgggact 1620
tgtggggtcc ttctcctgtc actggttatc accctttact gcaaacgggg cagaaagaaa 1680
ctcctgtata tattcaaaca accatttatg agaccagtac aaactactca agaggaagat 1740
ggctgtagct gccgatttcc agaagaagaa gaaggaggat gtgaactgag agtgaagttc 1800
agcaggagcg cagacgcccc cgcgtacaag cagggccaga accagctcta taacgagctc 1860
aatctaggac gaagagagga gtacgatgtt ttggacaaga gacgtggccg ggaccctgag 1920
atggggggaa agccgagaag gaagaaccct caggaaggcc tgtacaatga actgcagaaa 1980
gataagatgg cggaggccta cagtgagatt gggatgaaag gcgagcgccg gaggggcaag 2040
gggcacgatg gcctttacca gggtctcagt acagccacca aggacaccta cgacgccctt 2100
cacatgcagg ccctgccccc tcgc 2124
<210> 34
<211> 2895
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
gacatccaga tgacacagtc ccccagctcc ctgtctgcca gcgtgggcga ccgggtgacc 60
atcacatgca gagcctctca ggatatcagc aagtatctga actggtacca gcagaagcca 120
ggcaaggccc ccaggctgct gatctatcac acctcccgcc tgcactctgg agtgccaagc 180
cggttctccg gatctggaag cggaaccgac tacaccctga caatctctag cctgcagcct 240
gaggatttcg ccacatacta ttgccagcag ggcaataccc tgccatatac atttggcgga 300
ggaaccaggc tggagatcaa gggatccaca tctggaagcg gcaagccagg atccggagag 360
ggatctacca agggacaggt gcagctgcag gagtccggac ctggactggt gaagccaagc 420
cagacactgt ccctgacctg tacagtgagc ggcgtgtccc tgcctgatta cggcgtgtcc 480
tggatcagac agccacctgg caaggccctg gagtggctgg gcgtgatctg gggctctgag 540
accacatact attccacctc tctgaagacc aggctgacaa tctctaagga caacagcaag 600
aatcaggtgg tgctgaccat gacaaacatg gaccctgtgg ataccgccac atactattgt 660
gccaagcact actattacgg cggcagctat gccatggatt actggggcca gggctcctct 720
gtgaccgtga gctccacaac cacgcctgct ccacggcctc ctacccccgc accgactatt 780
gccagccaac cactcagcct gcggcccgag gcctgtagac ctgctgctgg cggggctgtc 840
cacacaagag gcctggattt tgcgtgcgac atttacatat gggcaccact cgcaggaacc 900
tgcggcgtgc tcctgctttc cctggtgatc acactgtact gtaagagagg gcggaaaaag 960
ctgctctaca tctttaagca gcccttcatg cgacctgttc agaccaccca ggaagaggac 1020
ggatgcagtt gcagattccc tgaggaagag gaaggcggct gtgagctcag ggtgaaattc 1080
tccagaagtg ccgatgcacc tgcctataaa caggggcaga atcagctgta caacgaactg 1140
aacctcggca gacgagaaga atatgacgtg cttgacaagc gtcggggaag agatccagaa 1200
atgggcggga agcccaggcg caagaatcca caggagggtc tctacaacga gcttcagaag 1260
gacaaaatgg ccgaagcgta cagcgagatc ggcatgaagg gagagagacg cagaggcaaa 1320
ggccacgacg gactgtacca aggactgagc accgcgacaa aggatactta cgatgcgctg 1380
cacatgcaag cgcttccgcc acggggaagc ggagctacta acttcagcct gctgaagcag 1440
gctggagacg tggaggagaa ccctggacct gaaacgacac tcacgcagtc tccagccacc 1500
ctgtctgtgt ctccagggga aagagccacc ctctcctgca gggccagtca gagtgttagc 1560
agcaacttag cctggtacca gcagaaacct ggccaggctc ccagactcct catctatggt 1620
gcatccacca gggccactgg catcccagcc aggttcagtg gcagtgggtc tgggaccgag 1680
ttcactctca ccatcagcag cctgcagtct gaggattttg caatttatta ctgtcagcag 1740
tatcatagct ggcctcccct cactttcggc ggagggacca agctggagat caaacgtgga 1800
agcaccagcg gcagcggaaa gcctggcagc ggcgagggca gcacaaaagg acaggtccag 1860
ctggtacagt ctggagctga ggtgaagaag cctggggcct cagtgaaggt ctcctgcaag 1920
gcttctggtt acacctttac cagctatggt atcagctggg tgcgacaggc ccctggacaa 1980
gggcttgagt ggatgggatg gatcagcgct tacaatggta acacaaacta tgcacagaag 2040
ctccagggca gagtcaccat gaccacagac acatccacga gcacagccta catggagctg 2100
aggagcctga gatctgacga cacggccgtg tattactgtg cgagagactt ccagggtata 2160
gcagtggctg atcttgacta ctggggccag ggaaccctgg tcacagtctc ctcaaagccc 2220
accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg 2280
tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg 2340
gacttcgcct gcgacttctg ggtgctggtc gtggtgggtg gcgtgctggc ctgctacagc 2400
ctgctggtga cagtggcctt catcatcttt tgggtgagga gcaagcggag cagaggcggc 2460
cacagcgact acatgaacat gaccccccgg aggcctggcc ccacccggaa gcactaccag 2520
ccctacgccc ctcccaggga cttcgccgcc taccggagcc gggtgaagtt cagccggagc 2580
gccgacgccc ctgcctacca gcagggccag agccagctgt acaacgagct gaacctgggc 2640
cggagggagg agtacgacgt gctggacaag cggagaggcc gggaccctga gatgggcggc 2700
aagccccgga gaaagaaccc tcaggagggc ctgtataacg aactgcagaa agacaagatg 2760
gccgaggcct acagcgagat cggcatgaag ggcgagcggc ggaggggcaa gggccacgac 2820
ggcctgtacc agggcctgag caccgccacc aaggatacct acgacgccct gcacatgcag 2880
gccctgcccc ctcgc 2895
<210> 35
<211> 327
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 240
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 300
ggagggacca agctggagat caaacgt 327
<210> 36
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
caggtccagc tggtacagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60
tcctgcaagg cttctggtta cacctttacc agctatggta tcagctgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatgg atcagcgctt acaatggtaa cacaaactat 180
gcacagaagc tccagggcag agtcaccatg accacagaca catccacgag cacagcctac 240
atggagctga ggagcctgag atctgacgac acggccgtgt attactgtgc gagagacttc 300
cagggtatag cagtggctga tcttgactac tggggccagg gaaccctggt cacagtctcc 360
tca 363
<210> 37
<211> 321
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
gacatccaga tgacacagtc ccccagctcc ctgtctgcca gcgtgggcga ccgggtgacc 60
atcacatgca gagcctctca ggatatcagc aagtatctga actggtacca gcagaagcca 120
ggcaaggccc ccaggctgct gatctatcac acctcccgcc tgcactctgg agtgccaagc 180
cggttctccg gatctggaag cggaaccgac tacaccctga caatctctag cctgcagcct 240
gaggatttcg ccacatacta ttgccagcag ggcaataccc tgccatatac atttggcgga 300
ggaaccaggc tggagatcaa g 321
<210> 38
<211> 360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
caggtgcagc tgcaggagtc cggacctgga ctggtgaagc caagccagac actgtccctg 60
acctgtacag tgagcggcgt gtccctgcct gattacggcg tgtcctggat cagacagcca 120
cctggcaagg ccctggagtg gctgggcgtg atctggggct ctgagaccac atactattcc 180
acctctctga agaccaggct gacaatctct aaggacaaca gcaagaatca ggtggtgctg 240
accatgacaa acatggaccc tgtggatacc gccacatact attgtgccaa gcactactat 300
tacggcggca gctatgccat ggattactgg ggccagggct cctctgtgac cgtgagctcc 360
<210> 39
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
ggaagcacca gcggcagcgg aaagcctggc agcggcgagg gcagcacaaa agga 54
<210> 40
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 40
ggatccacat ctggaagcgg caagccagga tccggagagg gatctaccaa ggga 54
<210> 41
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
gaggccgctg caaaggaagc agctgccaaa gaggctgcag ccaag 45
<210> 42
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
ggaggcggcg gttcaggtgg tggcggatct ggcggaggtg gttccggagg tggaggttca 60
<210> 43
<211> 15
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
gggggtggag gctct 15
<210> 44
<211> 15
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
ggtggaggcg ggtct 15
<210> 45
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct 60
ggacct 66
<210> 46
<211> 135
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg 60
tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg 120
gacttcgcct gtgat 135
<210> 47
<211> 134
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
caaccacgcc tgctccacgg cctcctaccc ccgcaccgac tattgccagc caaccactca 60
gcctgcggcc cgaggcctgt agacctgctg ctggcggggc tgtccacaca agaggcctgg 120
attttgcgtg cgac 134
<210> 48
<211> 141
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
aagcccacca cgacgccagc gccgcgacca ccaacaccgg cgcccaccat cgcgtcgcag 60
cccctgtccc tgcgcccaga ggcgtgccgg ccagcggcgg ggggcgcagt gcacacgagg 120
gggctggact tcgcctgcga c 141
<210> 49
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
atctacatct gggcgccctt ggccgggact tgtggggtcc ttctcctgtc actggttatc 60
accctttact gc 72
<210> 50
<211> 72
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
atttacatat gggcaccact cgcaggaacc tgcggcgtgc tcctgctttc cctggtgatc 60
acactgtact gt 72
<210> 51
<211> 81
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
ttctgggtgc tggtcgtggt gggtggcgtg ctggcctgct acagcctgct ggtgacagtg 60
gccttcatca tcttttgggt g 81
<210> 52
<211> 336
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
agagtgaagt tcagcaggag cgcagacgcc cccgcgtaca agcagggcca gaaccagctc 60
tataacgagc tcaatctagg acgaagagag gagtacgatg ttttggacaa gagacgtggc 120
cgggaccctg agatgggggg aaagccgaga aggaagaacc ctcaggaagg cctgtacaat 180
gaactgcaga aagataagat ggcggaggcc tacagtgaga ttgggatgaa aggcgagcgc 240
cggaggggca aggggcacga tggcctttac cagggtctca gtacagccac caaggacacc 300
tacgacgccc ttcacatgca ggccctgccc cctcgc 336
<210> 53
<211> 126
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
aaacggggca gaaagaaact cctgtatata ttcaaacaac catttatgag accagtacaa 60
actactcaag aggaagatgg ctgtagctgc cgatttccag aagaagaaga aggaggatgt 120
gaactg 126
<210> 54
<211> 336
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
agggtgaaat tctccagaag tgccgatgca cctgcctata aacaggggca gaatcagctg 60
tacaacgaac tgaacctcgg cagacgagaa gaatatgacg tgcttgacaa gcgtcgggga 120
agagatccag aaatgggcgg gaagcccagg cgcaagaatc cacaggaggg tctctacaac 180
gagcttcaga aggacaaaat ggccgaagcg tacagcgaga tcggcatgaa gggagagaga 240
cgcagaggca aaggccacga cggactgtac caaggactga gcaccgcgac aaaggatact 300
tacgatgcgc tgcacatgca agcgcttccg ccacgg 336
<210> 55
<211> 126
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
aagagagggc ggaaaaagct gctctacatc tttaagcagc ccttcatgcg acctgttcag 60
accacccagg aagaggacgg atgcagttgc agattccctg aggaagagga aggcggctgt 120
gagctc 126
<210> 56
<211> 336
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
cgggtgaagt tcagccggag cgccgacgcc cctgcctacc agcagggcca gagccagctg 60
tacaacgagc tgaacctggg ccggagggag gagtacgacg tgctggacaa gcggagaggc 120
cgggaccctg agatgggcgg caagccccgg agaaagaacc ctcaggaggg cctgtataac 180
gaactgcaga aagacaagat ggccgaggcc tacagcgaga tcggcatgaa gggcgagcgg 240
cggaggggca agggccacga cggcctgtac cagggcctga gcaccgccac caaggatacc 300
tacgacgccc tgcacatgca ggccctgccc cctcgc 336
<210> 57
<211> 123
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
aggagcaagc ggagcagagg cggccacagc gactacatga acatgacccc ccggaggcct 60
ggccccaccc ggaagcacta ccagccctac gcccctccca gggacttcgc cgcctaccgg 120
agc 123
<210> 58
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
atggccctgc ccgtgaccgc actcctgctg ccactggctc tcctgctgca cgctgcaagg 60
cct 63
<210> 59
<211> 18
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro
1 5 10 15
Gly Pro
<210> 60
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Gln Cys Thr Asn Tyr Ala Leu Leu Lys Leu Ala Gly Asp Val Glu Ser
1 5 10 15
Asn Pro Gly Pro
20
<210> 61
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg ccca 54
<210> 62
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
caatgtacta actacgcttt gttgaaactc gctggcgatg ttgaaagtaa ccccggtcct 60
<210> 63
<211> 278
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
atcccacagt gcatacgtgg gctccaacag gtcctcttgt cgagccacag tgcatacgtg 60
ggctccaaca ggtcctcttg tcgagccaca gtgcatacgt gggctccaac aggtcctctt 120
gtcgagccac agtgcatacg tgggctccaa caggtcctct tgtcgagcca cagtgcatac 180
gtgggctcca acaggtcctc ttgtcgagat ctggtaggcg tgtacggtgg gaggtctata 240
taagcagagc tcgtttagtg aaccgtcaga tcactagg 278
<210> 64
<211> 63
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
atggcactgc ctgtgaccgc cctgctgctg ccactggccc tgctgctgca cgcagcaagg 60
cca 63
<210> 65
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
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 Thr Ile Trp Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Asn Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Arg Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Ile Pro Gln
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 66
<211> 124
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn
20 25 30
Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu
35 40 45
Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala
50 55 60
Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn
65 70 75 80
Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val
85 90 95
Tyr Tyr Cys Ala Arg Glu Val Thr Gly Asp Leu Glu Asp Ala Phe Asp
100 105 110
Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser
115 120
<210> 67
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
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 Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 68
<211> 120
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ser Leu Lys
50 55 60
Thr Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu
65 70 75 80
Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Ser Ser Val Thr Val Ser Ser
115 120
<210> 69
<211> 321
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
gatatccaga tgacacagtc cccctcctct ctgagcgcct ccgtgggcga cagggtgacc 60
atcacatgcc gcgcctctca gaccatctgg agctacctga actggtatca gcagaggcca 120
ggcaaggcac ctaatctgct gatctacgca gccagctccc tgcagtccgg agtgccttct 180
aggttcagcg gaaggggatc tggaaccgac ttcaccctga caatctctag cctgcaggcc 240
gaggacttcg ccacatacta ttgtcagcag tcttatagca tcccacagac ctttggccag 300
ggcacaaagc tggagatcaa g 321
<210> 70
<211> 372
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
caggtgcagc tgcagcagag cggaccagga ctggtgaagc catcccagac cctgtctctg 60
acatgcgcca tctccggcga ttctgtgagc tccaacagcg ccgcctggaa ttggatccgg 120
cagtccccca gccggggcct ggagtggctg ggacggacat actatagatc caagtggtac 180
aacgattatg ccgtgagcgt gaagtcccgg atcaccatca accccgacac atctaagaat 240
cagttcagcc tgcagctgaa cagcgtgacc cctgaggaca cagccgtgta ctattgtgcc 300
agagaggtga ccggcgacct ggaggatgcc tttgacatct ggggccaggg caccatggtg 360
acagtgtcta gc 372
<210> 71
<211> 321
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
gacatccaga tgacccagag cccttcttct ctgagcgcca gcgtgggaga cagagtgacc 60
atcacttgca gggccagcca ggacatcagc aagtacctga attggtacca gcagaagcca 120
ggcaaggccc ctagactgct gatctaccac acaagcagac tgcacagcgg agtgcctagc 180
agattcagcg gcagcggaag cggaaccgac tacaccctga ccatcagcag cctgcagcca 240
gaggacttcg ccacctacta ctgccagcag ggcaacacac tgccttacac cttcggcgga 300
ggcacaagac tggagatcaa g 321
<210> 72
<211> 360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
caggtgcagc tgcaggaaag cggaccagga ctggtgaagc cttctcagac cctgagcctg 60
acttgcaccg tgtcaggagt gtccctgcca gattacggcg tgtcttggat cagacagccc 120
ccaggaaagg ccctggagtg gctgggagtg atttggggaa gcgagaccac ctactacaac 180
agcagcctga agacccggct gaccatcagc aaggacaaca gcaagaacca ggtggtgctg 240
accatgacca acatggaccc cgtggacacc gccacctact attgcgccaa gcactactac 300
tacggcggaa gctacgccat ggactattgg ggccagggaa gcagcgtgac cgtgtctagc 360
<210> 73
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
ggcagcacaa gcggaagcgg caaaccagga agcggagaag gaagcaccaa ggga 54
<210> 74
<211> 486
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn
20 25 30
Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu
35 40 45
Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala
50 55 60
Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn
65 70 75 80
Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val
85 90 95
Tyr Tyr Cys Ala Arg Glu Val Thr Gly Asp Leu Glu Asp Ala Phe Asp
100 105 110
Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly
115 120 125
Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
130 135 140
Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys
145 150 155 160
Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu
165 170 175
Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser
180 185 190
Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln
195 200 205
Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro
210 215 220
Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser
225 230 235 240
Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val
245 250 255
Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu
260 265 270
Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val
275 280 285
Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val
290 295 300
Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg
305 310 315 320
Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met
325 330 335
Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His
340 345 350
Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser
355 360 365
Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr
370 375 380
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile
385 390 395 400
Thr Cys Arg Ala Ser Gln Thr Ile Trp Ser Tyr Leu Asn Trp Tyr Gln
405 410 415
Gln Arg Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr Ala Ala Ser Ser
420 425 430
Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Arg Gly Ser Gly Thr
435 440 445
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Phe Ala Thr
450 455 460
Tyr Tyr Cys Gln Gln Ser Tyr Ser Ile Pro Gln Thr Phe Gly Gln Gly
465 470 475 480
Thr Lys Leu Glu Ile Lys
485
<210> 75
<211> 508
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Tyr His Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Thr
100 105 110
Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln
115 120 125
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser
130 135 140
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly
145 150 155 160
Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
165 170 175
Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln
180 185 190
Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met
195 200 205
Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln
225 230 235 240
Gly Thr Leu Val Thr Val Ser Ser Glu Ala Ala Ala Lys Glu Ala Ala
245 250 255
Ala Lys Glu Ala Ala Ala Lys Asp Ile Gln Met Thr Gln Ser Pro Ser
260 265 270
Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala
275 280 285
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly
290 295 300
Lys Ala Pro Arg Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
305 310 315 320
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu
325 330 335
Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
340 345 350
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu
355 360 365
Ile Lys Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly
370 375 380
Ser Thr Lys Gly Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val
385 390 395 400
Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser
405 410 415
Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala
420 425 430
Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn
435 440 445
Ser Ser Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn
450 455 460
Gln Val Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr
465 470 475 480
Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp
485 490 495
Tyr Trp Gly Gln Gly Ser Ser Val Thr Val Ser Ser
500 505
<210> 76
<211> 709
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn
20 25 30
Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu
35 40 45
Trp Leu Gly Arg Thr Tyr Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala
50 55 60
Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn
65 70 75 80
Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val
85 90 95
Tyr Tyr Cys Ala Arg Glu Val Thr Gly Asp Leu Glu Asp Ala Phe Asp
100 105 110
Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly
115 120 125
Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
130 135 140
Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Lys
145 150 155 160
Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu
165 170 175
Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser
180 185 190
Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln
195 200 205
Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro
210 215 220
Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser
225 230 235 240
Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val
245 250 255
Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu
260 265 270
Ser Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val
275 280 285
Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val
290 295 300
Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg
305 310 315 320
Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met
325 330 335
Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His
340 345 350
Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser
355 360 365
Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr
370 375 380
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile
385 390 395 400
Thr Cys Arg Ala Ser Gln Thr Ile Trp Ser Tyr Leu Asn Trp Tyr Gln
405 410 415
Gln Arg Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr Ala Ala Ser Ser
420 425 430
Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Arg Gly Ser Gly Thr
435 440 445
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Phe Ala Thr
450 455 460
Tyr Tyr Cys Gln Gln Ser Tyr Ser Ile Pro Gln Thr Phe Gly Gln Gly
465 470 475 480
Thr Lys Leu Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr
485 490 495
Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala
500 505 510
Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe
515 520 525
Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val
530 535 540
Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys
545 550 555 560
Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr
565 570 575
Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu
580 585 590
Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro
595 600 605
Ala Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly
610 615 620
Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro
625 630 635 640
Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr
645 650 655
Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly
660 665 670
Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln
675 680 685
Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln
690 695 700
Ala Leu Pro Pro Arg
705
<210> 77
<211> 731
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Tyr His Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Gly Ser Thr
100 105 110
Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln
115 120 125
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser
130 135 140
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Gly
145 150 155 160
Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
165 170 175
Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln
180 185 190
Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met
195 200 205
Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Asp Phe Gln Gly Ile Ala Val Ala Asp Leu Asp Tyr Trp Gly Gln
225 230 235 240
Gly Thr Leu Val Thr Val Ser Ser Glu Ala Ala Ala Lys Glu Ala Ala
245 250 255
Ala Lys Glu Ala Ala Ala Lys Asp Ile Gln Met Thr Gln Ser Pro Ser
260 265 270
Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala
275 280 285
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly
290 295 300
Lys Ala Pro Arg Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
305 310 315 320
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu
325 330 335
Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
340 345 350
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Arg Leu Glu
355 360 365
Ile Lys Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly
370 375 380
Ser Thr Lys Gly Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val
385 390 395 400
Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser
405 410 415
Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Gly Lys Ala
420 425 430
Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn
435 440 445
Ser Ser Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn
450 455 460
Gln Val Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr
465 470 475 480
Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp
485 490 495
Tyr Trp Gly Gln Gly Ser Ser Val Thr Val Ser Ser Thr Thr Thr Pro
500 505 510
Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu
515 520 525
Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His
530 535 540
Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu
545 550 555 560
Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr
565 570 575
Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe
580 585 590
Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg
595 600 605
Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser
610 615 620
Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln Leu Tyr
625 630 635 640
Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys
645 650 655
Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn
660 665 670
Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu
675 680 685
Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly
690 695 700
His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr
705 710 715 720
Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg
725 730
<210> 78
<211> 1458
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
caggtgcagc tgcagcagag cggaccagga ctggtgaagc catcccagac cctgtctctg 60
acatgcgcca tctccggcga ttctgtgagc tccaacagcg ccgcctggaa ttggatccgg 120
cagtccccca gccggggcct ggagtggctg ggacggacat actatagatc caagtggtac 180
aacgattatg ccgtgagcgt gaagtcccgg atcaccatca accccgacac atctaagaat 240
cagttcagcc tgcagctgaa cagcgtgacc cctgaggaca cagccgtgta ctattgtgcc 300
agagaggtga ccggcgacct ggaggatgcc tttgacatct ggggccaggg caccatggtg 360
acagtgtcta gcgggggtgg aggctctgac atccagatga cacagtcccc cagctccctg 420
tctgccagcg tgggcgaccg ggtgaccatc acatgcagag cctctcagga tatcagcaag 480
tatctgaact ggtaccagca gaagccaggc aaggccccca ggctgctgat ctatcacacc 540
tcccgcctgc actctggagt gccaagccgg ttctccggat ctggaagcgg aaccgactac 600
accctgacaa tctctagcct gcagcctgag gatttcgcca catactattg ccagcagggc 660
aataccctgc catatacatt tggcggagga accaggctgg agatcaaggg atccacatct 720
ggaagcggca agccaggatc cggagaggga tctaccaagg gacaggtgca gctgcaggag 780
tccggacctg gactggtgaa gccaagccag acactgtccc tgacctgtac agtgagcggc 840
gtgtccctgc ctgattacgg cgtgtcctgg atcagacagc cacctggcaa ggccctggag 900
tggctgggcg tgatctgggg ctctgagacc acatactatt ccacctctct gaagaccagg 960
ctgacaatct ctaaggacaa cagcaagaat caggtggtgc tgaccatgac aaacatggac 1020
cctgtggata ccgccacata ctattgtgcc aagcactact attacggcgg cagctatgcc 1080
atggattact ggggccaggg ctcctctgtg accgtgagct ccggtggagg cgggtctgat 1140
atccagatga cacagtcccc ctcctctctg agcgcctccg tgggcgacag ggtgaccatc 1200
acatgccgcg cctctcagac catctggagc tacctgaact ggtatcagca gaggccaggc 1260
aaggcaccta atctgctgat ctacgcagcc agctccctgc agtccggagt gccttctagg 1320
ttcagcggaa ggggatctgg aaccgacttc accctgacaa tctctagcct gcaggccgag 1380
gacttcgcca catactattg tcagcagtct tatagcatcc cacagacctt tggccagggc 1440
acaaagctgg agatcaag 1458
<210> 79
<211> 1524
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 240
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 300
ggagggacca agctggagat caaacgtgga agcaccagcg gcagcggaaa gcctggcagc 360
ggcgagggca gcacaaaagg acaggtccag ctggtacagt ctggagctga ggtgaagaag 420
cctggggcct cagtgaaggt ctcctgcaag gcttctggtt acacctttac cagctatggt 480
atcagctggg tgcgacaggc ccctggacaa gggcttgagt ggatgggatg gatcagcgct 540
tacaatggta acacaaacta tgcacagaag ctccagggca gagtcaccat gaccacagac 600
acatccacga gcacagccta catggagctg aggagcctga gatctgacga cacggccgtg 660
tattactgtg cgagagactt ccagggtata gcagtggctg atcttgacta ctggggccag 720
ggaaccctgg tcacagtctc ctcagaggcc gctgcaaagg aagcagctgc caaagaggct 780
gcagccaagg acatccagat gacccagagc ccttcttctc tgagcgccag cgtgggagac 840
agagtgacca tcacttgcag ggccagccag gacatcagca agtacctgaa ttggtaccag 900
cagaagccag gcaaggcccc tagactgctg atctaccaca caagcagact gcacagcgga 960
gtgcctagca gattcagcgg cagcggaagc ggaaccgact acaccctgac catcagcagc 1020
ctgcagccag aggacttcgc cacctactac tgccagcagg gcaacacact gccttacacc 1080
ttcggcggag gcacaagact ggagatcaag ggcagcacaa gcggaagcgg caaaccagga 1140
agcggagaag gaagcaccaa gggacaggtg cagctgcagg aaagcggacc aggactggtg 1200
aagccttctc agaccctgag cctgacttgc accgtgtcag gagtgtccct gccagattac 1260
ggcgtgtctt ggatcagaca gcccccagga aaggccctgg agtggctggg agtgatttgg 1320
ggaagcgaga ccacctacta caacagcagc ctgaagaccc ggctgaccat cagcaaggac 1380
aacagcaaga accaggtggt gctgaccatg accaacatgg accccgtgga caccgccacc 1440
tactattgcg ccaagcacta ctactacggc ggaagctacg ccatggacta ttggggccag 1500
ggaagcagcg tgaccgtgtc tagc 1524
<210> 80
<211> 2130
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
caggtgcagc tgcagcagag cggaccagga ctggtgaagc catcccagac cctgtctctg 60
acatgcgcca tctccggcga ttctgtgagc tccaacagcg ccgcctggaa ttggatccgg 120
cagtccccca gccggggcct ggagtggctg ggacggacat actatagatc caagtggtac 180
aacgattatg ccgtgagcgt gaagtcccgg atcaccatca accccgacac atctaagaat 240
cagttcagcc tgcagctgaa cagcgtgacc cctgaggaca cagccgtgta ctattgtgcc 300
agagaggtga ccggcgacct ggaggatgcc tttgacatct ggggccaggg caccatggtg 360
acagtgtcta gcgggggtgg aggctctgac atccagatga cacagtcccc cagctccctg 420
tctgccagcg tgggcgaccg ggtgaccatc acatgcagag cctctcagga tatcagcaag 480
tatctgaact ggtaccagca gaagccaggc aaggccccca ggctgctgat ctatcacacc 540
tcccgcctgc actctggagt gccaagccgg ttctccggat ctggaagcgg aaccgactac 600
accctgacaa tctctagcct gcagcctgag gatttcgcca catactattg ccagcagggc 660
aataccctgc catatacatt tggcggagga accaggctgg agatcaaggg atccacatct 720
ggaagcggca agccaggatc cggagaggga tctaccaagg gacaggtgca gctgcaggag 780
tccggacctg gactggtgaa gccaagccag acactgtccc tgacctgtac agtgagcggc 840
gtgtccctgc ctgattacgg cgtgtcctgg atcagacagc cacctggcaa ggccctggag 900
tggctgggcg tgatctgggg ctctgagacc acatactatt ccacctctct gaagaccagg 960
ctgacaatct ctaaggacaa cagcaagaat caggtggtgc tgaccatgac aaacatggac 1020
cctgtggata ccgccacata ctattgtgcc aagcactact attacggcgg cagctatgcc 1080
atggattact ggggccaggg ctcctctgtg accgtgagct ccggtggagg cgggtctgat 1140
atccagatga cacagtcccc ctcctctctg agcgcctccg tgggcgacag ggtgaccatc 1200
acatgccgcg cctctcagac catctggagc tacctgaact ggtatcagca gaggccaggc 1260
aaggcaccta atctgctgat ctacgcagcc agctccctgc agtccggagt gccttctagg 1320
ttcagcggaa ggggatctgg aaccgacttc accctgacaa tctctagcct gcaggccgag 1380
gacttcgcca catactattg tcagcagtct tatagcatcc cacagacctt tggccagggc 1440
acaaagctgg agatcaagac cacgacgcca gcgccgcgac caccaacacc ggcgcccacc 1500
atcgcgtcgc agcccctgtc cctgcgccca gaggcgtgcc ggccagcggc ggggggcgca 1560
gtgcacacga gggggctgga cttcgcctgt gatatctaca tctgggcgcc cttggccggg 1620
acttgtgggg tccttctcct gtcactggtt atcacccttt actgcaaacg gggcagaaag 1680
aaactcctgt atatattcaa acaaccattt atgagaccag tacaaactac tcaagaggaa 1740
gatggctgta gctgccgatt tccagaagaa gaagaaggag gatgtgaact gagagtgaag 1800
ttcagcagga gcgcagacgc ccccgcgtac aagcagggcc agaaccagct ctataacgag 1860
ctcaatctag gacgaagaga ggagtacgat gttttggaca agagacgtgg ccgggaccct 1920
gagatggggg gaaagccgag aaggaagaac cctcaggaag gcctgtacaa tgaactgcag 1980
aaagataaga tggcggaggc ctacagtgag attgggatga aaggcgagcg ccggaggggc 2040
aaggggcacg atggccttta ccagggtctc agtacagcca ccaaggacac ctacgacgcc 2100
cttcacatgc aggccctgcc ccctcgctaa 2130
<210> 81
<211> 2196
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
gaaacgacac tcacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120
ggccaggctc ccagactcct catctatggt gcatccacca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggaccgag ttcactctca ccatcagcag cctgcagtct 240
gaggattttg caatttatta ctgtcagcag tatcatagct ggcctcccct cactttcggc 300
ggagggacca agctggagat caaacgtgga agcaccagcg gcagcggaaa gcctggcagc 360
ggcgagggca gcacaaaagg acaggtccag ctggtacagt ctggagctga ggtgaagaag 420
cctggggcct cagtgaaggt ctcctgcaag gcttctggtt acacctttac cagctatggt 480
atcagctggg tgcgacaggc ccctggacaa gggcttgagt ggatgggatg gatcagcgct 540
tacaatggta acacaaacta tgcacagaag ctccagggca gagtcaccat gaccacagac 600
acatccacga gcacagccta catggagctg aggagcctga gatctgacga cacggccgtg 660
tattactgtg cgagagactt ccagggtata gcagtggctg atcttgacta ctggggccag 720
ggaaccctgg tcacagtctc ctcagaggcc gctgcaaagg aagcagctgc caaagaggct 780
gcagccaagg acatccagat gacccagagc ccttcttctc tgagcgccag cgtgggagac 840
agagtgacca tcacttgcag ggccagccag gacatcagca agtacctgaa ttggtaccag 900
cagaagccag gcaaggcccc tagactgctg atctaccaca caagcagact gcacagcgga 960
gtgcctagca gattcagcgg cagcggaagc ggaaccgact acaccctgac catcagcagc 1020
ctgcagccag aggacttcgc cacctactac tgccagcagg gcaacacact gccttacacc 1080
ttcggcggag gcacaagact ggagatcaag ggcagcacaa gcggaagcgg caaaccagga 1140
agcggagaag gaagcaccaa gggacaggtg cagctgcagg aaagcggacc aggactggtg 1200
aagccttctc agaccctgag cctgacttgc accgtgtcag gagtgtccct gccagattac 1260
ggcgtgtctt ggatcagaca gcccccagga aaggccctgg agtggctggg agtgatttgg 1320
ggaagcgaga ccacctacta caacagcagc ctgaagaccc ggctgaccat cagcaaggac 1380
aacagcaaga accaggtggt gctgaccatg accaacatgg accccgtgga caccgccacc 1440
tactattgcg ccaagcacta ctactacggc ggaagctacg ccatggacta ttggggccag 1500
ggaagcagcg tgaccgtgtc tagcaccacg acgccagcgc cgcgaccacc aacaccggcg 1560
cccaccatcg cgtcgcagcc cctgtccctg cgcccagagg cgtgccggcc agcggcgggg 1620
ggcgcagtgc acacgagggg gctggacttc gcctgtgata tctacatctg ggcgcccttg 1680
gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg caaacggggc 1740
agaaagaaac tcctgtatat attcaaacaa ccatttatga gaccagtaca aactactcaa 1800
gaggaagatg gctgtagctg ccgatttcca gaagaagaag aaggaggatg tgaactgaga 1860
gtgaagttca gcaggagcgc agacgccccc gcgtacaagc agggccagaa ccagctctat 1920
aacgagctca atctaggacg aagagaggag tacgatgttt tggacaagag acgtggccgg 1980
gaccctgaga tggggggaaa gccgagaagg aagaaccctc aggaaggcct gtacaatgaa 2040
ctgcagaaag ataagatggc ggaggcctac agtgagattg ggatgaaagg cgagcgccgg 2100
aggggcaagg ggcacgatgg cctttaccag ggtctcagta cagccaccaa ggacacctac 2160
gacgcccttc acatgcaggc cctgccccct cgctaa 2196
<210> 82
<211> 961
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
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 Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln
115 120 125
Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser
130 135 140
Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser
145 150 155 160
Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile
165 170 175
Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu
180 185 190
Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr
195 200 205
Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr
210 215 220
Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser
225 230 235 240
Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
245 250 255
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
260 265 270
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
275 280 285
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
290 295 300
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys
305 310 315 320
Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr
325 330 335
Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly
340 345 350
Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala
355 360 365
Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg
370 375 380
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu
385 390 395 400
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn
405 410 415
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met
420 425 430
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly
435 440 445
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala
450 455 460
Leu Pro Pro Arg Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val
465 470 475 480
Glu Glu Asn Pro Gly Pro Glu Thr Thr Leu Thr Gln Ser Pro Ala Thr
485 490 495
Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser
500 505 510
Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
515 520 525
Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile
530 535 540
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr
545 550 555 560
Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln
565 570 575
Tyr His Ser Trp Pro Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
580 585 590
Ile Lys Arg Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu
595 600 605
Gly Ser Thr Lys Gly Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
610 615 620
Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
625 630 635 640
Thr Phe Thr Ser Tyr Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln
645 650 655
Gly Leu Glu Trp Met Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn
660 665 670
Tyr Ala Gln Lys Leu Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser
675 680 685
Thr Ser Thr Ala Tyr Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr
690 695 700
Ala Val Tyr Tyr Cys Ala Arg Asp Phe Gln Gly Ile Ala Val Ala Asp
705 710 715 720
Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Lys Pro
725 730 735
Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala
740 745 750
Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly
755 760 765
Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Phe Trp Val
770 775 780
Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr
785 790 795 800
Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Gly Gly
805 810 815
His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg
820 825 830
Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg
835 840 845
Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln
850 855 860
Gly Gln Ser Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu
865 870 875 880
Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly
885 890 895
Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln
900 905 910
Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu
915 920 925
Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr
930 935 940
Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro
945 950 955 960
Arg
<210> 83
<211> 963
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
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 Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Arg Leu Glu Ile Lys Gly Ser Thr Ser Gly
100 105 110
Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Gln Val Gln
115 120 125
Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser
130 135 140
Leu Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr Gly Val Ser
145 150 155 160
Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu Gly Val Ile
165 170 175
Trp Gly Ser Glu Thr Thr Tyr Tyr Ser Thr Ser Leu Lys Thr Arg Leu
180 185 190
Thr Ile Ser Lys Asp Asn Ser Lys Asn Gln Val Val Leu Thr Met Thr
195 200 205
Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Lys His Tyr
210 215 220
Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Ser Ser
225 230 235 240
Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
245 250 255
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
260 265 270
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
275 280 285
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
290 295 300
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys
305 310 315 320
Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr
325 330 335
Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly
340 345 350
Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala
355 360 365
Tyr Lys Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg
370 375 380
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu
385 390 395 400
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn
405 410 415
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met
420 425 430
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly
435 440 445
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala
450 455 460
Leu Pro Pro Arg Gln Cys Thr Asn Tyr Ala Leu Leu Lys Leu Ala Gly
465 470 475 480
Asp Val Glu Ser Asn Pro Gly Pro Glu Thr Thr Leu Thr Gln Ser Pro
485 490 495
Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg
500 505 510
Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro
515 520 525
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr
530 535 540
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
545 550 555 560
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Ile Tyr Tyr Cys
565 570 575
Gln Gln Tyr His Ser Trp Pro Pro Leu Thr Phe Gly Gly Gly Thr Lys
580 585 590
Leu Glu Ile Lys Arg Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser
595 600 605
Gly Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Val Gln Ser Gly Ala
610 615 620
Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser
625 630 635 640
Gly Tyr Thr Phe Thr Ser Tyr Gly Ile Ser Trp Val Arg Gln Ala Pro
645 650 655
Gly Gln Gly Leu Glu Trp Met Gly Trp Ile Ser Ala Tyr Asn Gly Asn
660 665 670
Thr Asn Tyr Ala Gln Lys Leu Gln Gly Arg Val Thr Met Thr Thr Asp
675 680 685
Thr Ser Thr Ser Thr Ala Tyr Met Glu Leu Arg Ser Leu Arg Ser Asp
690 695 700
Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Phe Gln Gly Ile Ala Val
705 710 715 720
Ala Asp Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
725 730 735
Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr
740 745 750
Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala
755 760 765
Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Phe
770 775 780
Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu
785 790 795 800
Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg
805 810 815
Gly Gly His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro
820 825 830
Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala
835 840 845
Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr
850 855 860
Gln Gln Gly Gln Ser Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg
865 870 875 880
Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met
885 890 895
Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu
900 905 910
Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys
915 920 925
Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu
930 935 940
Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu
945 950 955 960
Pro Pro Arg
<210> 84
<211> 2883
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
gacatccaga tgacacagtc ccccagctcc ctgtctgcca gcgtgggcga ccgggtgacc 60
atcacatgca gagcctctca ggatatcagc aagtatctga actggtacca gcagaagcca 120
ggcaaggccc ccaggctgct gatctatcac acctcccgcc tgcactctgg agtgccaagc 180
cggttctccg gatctggaag cggaaccgac tacaccctga caatctctag cctgcagcct 240
gaggatttcg ccacatacta ttgccagcag ggcaataccc tgccatatac atttggcgga 300
ggaaccaggc tggagatcaa gggatccaca tctggaagcg gcaagccagg atccggagag 360
ggatctacca agggacaggt gcagctgcag gagtccggac ctggactggt gaagccaagc 420
cagacactgt ccctgacctg tacagtgagc ggcgtgtccc tgcctgatta cggcgtgtcc 480
tggatcagac agccacctgg caaggccctg gagtggctgg gcgtgatctg gggctctgag 540
accacatact attccacctc tctgaagacc aggctgacaa tctctaagga caacagcaag 600
aatcaggtgg tgctgaccat gacaaacatg gaccctgtgg ataccgccac atactattgt 660
gccaagcact actattacgg cggcagctat gccatggatt actggggcca gggctcctct 720
gtgaccgtga gctccacaac cacgcctgct ccacggcctc ctacccccgc accgactatt 780
gccagccaac cactcagcct gcggcccgag gcctgtagac ctgctgctgg cggggctgtc 840
cacacaagag gcctggattt tgcgtgcgac atttacatat gggcaccact cgcaggaacc 900
tgcggcgtgc tcctgctttc cctggtgatc acactgtact gtaagagagg gcggaaaaag 960
ctgctctaca tctttaagca gcccttcatg cgacctgttc agaccaccca ggaagaggac 1020
ggatgcagtt gcagattccc tgaggaagag gaaggcggct gtgagctcag ggtgaaattc 1080
tccagaagtg ccgatgcacc tgcctataaa caggggcaga atcagctgta caacgaactg 1140
aacctcggca gacgagaaga atatgacgtg cttgacaagc gtcggggaag agatccagaa 1200
atgggcggga agcccaggcg caagaatcca caggagggtc tctacaacga gcttcagaag 1260
gacaaaatgg ccgaagcgta cagcgagatc ggcatgaagg gagagagacg cagaggcaaa 1320
ggccacgacg gactgtacca aggactgagc accgcgacaa aggatactta cgatgcgctg 1380
cacatgcaag cgcttccgcc acgggagggc agaggaagtc tgctaacatg cggtgacgtc 1440
gaggagaatc ctggcccaga aacgacactc acgcagtctc cagccaccct gtctgtgtct 1500
ccaggggaaa gagccaccct ctcctgcagg gccagtcaga gtgttagcag caacttagcc 1560
tggtaccagc agaaacctgg ccaggctccc agactcctca tctatggtgc atccaccagg 1620
gccactggca tcccagccag gttcagtggc agtgggtctg ggaccgagtt cactctcacc 1680
atcagcagcc tgcagtctga ggattttgca atttattact gtcagcagta tcatagctgg 1740
cctcccctca ctttcggcgg agggaccaag ctggagatca aacgtggaag caccagcggc 1800
agcggaaagc ctggcagcgg cgagggcagc acaaaaggac aggtccagct ggtacagtct 1860
ggagctgagg tgaagaagcc tggggcctca gtgaaggtct cctgcaaggc ttctggttac 1920
acctttacca gctatggtat cagctgggtg cgacaggccc ctggacaagg gcttgagtgg 1980
atgggatgga tcagcgctta caatggtaac acaaactatg cacagaagct ccagggcaga 2040
gtcaccatga ccacagacac atccacgagc acagcctaca tggagctgag gagcctgaga 2100
tctgacgaca cggccgtgta ttactgtgcg agagacttcc agggtatagc agtggctgat 2160
cttgactact ggggccaggg aaccctggtc acagtctcct caaagcccac cacgacgcca 2220
gcgccgcgac caccaacacc ggcgcccacc atcgcgtcgc agcccctgtc cctgcgccca 2280
gaggcgtgcc ggccagcggc ggggggcgca gtgcacacga gggggctgga cttcgcctgc 2340
gacttctggg tgctggtcgt ggtgggtggc gtgctggcct gctacagcct gctggtgaca 2400
gtggccttca tcatcttttg ggtgaggagc aagcggagca gaggcggcca cagcgactac 2460
atgaacatga ccccccggag gcctggcccc acccggaagc actaccagcc ctacgcccct 2520
cccagggact tcgccgccta ccggagccgg gtgaagttca gccggagcgc cgacgcccct 2580
gcctaccagc agggccagag ccagctgtac aacgagctga acctgggccg gagggaggag 2640
tacgacgtgc tggacaagcg gagaggccgg gaccctgaga tgggcggcaa gccccggaga 2700
aagaaccctc aggagggcct gtataacgaa ctgcagaaag acaagatggc cgaggcctac 2760
agcgagatcg gcatgaaggg cgagcggcgg aggggcaagg gccacgacgg cctgtaccag 2820
ggcctgagca ccgccaccaa ggatacctac gacgccctgc acatgcaggc cctgccccct 2880
cgc 2883
<210> 85
<211> 2889
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
gacatccaga tgacacagtc ccccagctcc ctgtctgcca gcgtgggcga ccgggtgacc 60
atcacatgca gagcctctca ggatatcagc aagtatctga actggtacca gcagaagcca 120
ggcaaggccc ccaggctgct gatctatcac acctcccgcc tgcactctgg agtgccaagc 180
cggttctccg gatctggaag cggaaccgac tacaccctga caatctctag cctgcagcct 240
gaggatttcg ccacatacta ttgccagcag ggcaataccc tgccatatac atttggcgga 300
ggaaccaggc tggagatcaa gggatccaca tctggaagcg gcaagccagg atccggagag 360
ggatctacca agggacaggt gcagctgcag gagtccggac ctggactggt gaagccaagc 420
cagacactgt ccctgacctg tacagtgagc ggcgtgtccc tgcctgatta cggcgtgtcc 480
tggatcagac agccacctgg caaggccctg gagtggctgg gcgtgatctg gggctctgag 540
accacatact attccacctc tctgaagacc aggctgacaa tctctaagga caacagcaag 600
aatcaggtgg tgctgaccat gacaaacatg gaccctgtgg ataccgccac atactattgt 660
gccaagcact actattacgg cggcagctat gccatggatt actggggcca gggctcctct 720
gtgaccgtga gctccacaac cacgcctgct ccacggcctc ctacccccgc accgactatt 780
gccagccaac cactcagcct gcggcccgag gcctgtagac ctgctgctgg cggggctgtc 840
cacacaagag gcctggattt tgcgtgcgac atttacatat gggcaccact cgcaggaacc 900
tgcggcgtgc tcctgctttc cctggtgatc acactgtact gtaagagagg gcggaaaaag 960
ctgctctaca tctttaagca gcccttcatg cgacctgttc agaccaccca ggaagaggac 1020
ggatgcagtt gcagattccc tgaggaagag gaaggcggct gtgagctcag ggtgaaattc 1080
tccagaagtg ccgatgcacc tgcctataaa caggggcaga atcagctgta caacgaactg 1140
aacctcggca gacgagaaga atatgacgtg cttgacaagc gtcggggaag agatccagaa 1200
atgggcggga agcccaggcg caagaatcca caggagggtc tctacaacga gcttcagaag 1260
gacaaaatgg ccgaagcgta cagcgagatc ggcatgaagg gagagagacg cagaggcaaa 1320
ggccacgacg gactgtacca aggactgagc accgcgacaa aggatactta cgatgcgctg 1380
cacatgcaag cgcttccgcc acggcaatgt actaactacg ctttgttgaa actcgctggc 1440
gatgttgaaa gtaaccccgg tcctgaaacg acactcacgc agtctccagc caccctgtct 1500
gtgtctccag gggaaagagc caccctctcc tgcagggcca gtcagagtgt tagcagcaac 1560
ttagcctggt accagcagaa acctggccag gctcccagac tcctcatcta tggtgcatcc 1620
accagggcca ctggcatccc agccaggttc agtggcagtg ggtctgggac cgagttcact 1680
ctcaccatca gcagcctgca gtctgaggat tttgcaattt attactgtca gcagtatcat 1740
agctggcctc ccctcacttt cggcggaggg accaagctgg agatcaaacg tggaagcacc 1800
agcggcagcg gaaagcctgg cagcggcgag ggcagcacaa aaggacaggt ccagctggta 1860
cagtctggag ctgaggtgaa gaagcctggg gcctcagtga aggtctcctg caaggcttct 1920
ggttacacct ttaccagcta tggtatcagc tgggtgcgac aggcccctgg acaagggctt 1980
gagtggatgg gatggatcag cgcttacaat ggtaacacaa actatgcaca gaagctccag 2040
ggcagagtca ccatgaccac agacacatcc acgagcacag cctacatgga gctgaggagc 2100
ctgagatctg acgacacggc cgtgtattac tgtgcgagag acttccaggg tatagcagtg 2160
gctgatcttg actactgggg ccagggaacc ctggtcacag tctcctcaaa gcccaccacg 2220
acgccagcgc cgcgaccacc aacaccggcg cccaccatcg cgtcgcagcc cctgtccctg 2280
cgcccagagg cgtgccggcc agcggcgggg ggcgcagtgc acacgagggg gctggacttc 2340
gcctgcgact tctgggtgct ggtcgtggtg ggtggcgtgc tggcctgcta cagcctgctg 2400
gtgacagtgg ccttcatcat cttttgggtg aggagcaagc ggagcagagg cggccacagc 2460
gactacatga acatgacccc ccggaggcct ggccccaccc ggaagcacta ccagccctac 2520
gcccctccca gggacttcgc cgcctaccgg agccgggtga agttcagccg gagcgccgac 2580
gcccctgcct accagcaggg ccagagccag ctgtacaacg agctgaacct gggccggagg 2640
gaggagtacg acgtgctgga caagcggaga ggccgggacc ctgagatggg cggcaagccc 2700
cggagaaaga accctcagga gggcctgtat aacgaactgc agaaagacaa gatggccgag 2760
gcctacagcg agatcggcat gaagggcgag cggcggaggg gcaagggcca cgacggcctg 2820
taccagggcc tgagcaccgc caccaaggat acctacgacg ccctgcacat gcaggccctg 2880
ccccctcgc 2889

Claims (23)

1.靶向CD22和CD19的双特异性抗体，包括CD22轻链、CD22重链、CD19轻链、CD19重链，其特征在于，所述CD22轻链的可变区包含L-CDR1、L-CDR2和L-CDR3，所述CD22重链的可变区包含H-CDR1、H-CDR2和H-CDR3；
所述L-CDR1、L-CDR2和L-CDR3选自以下氨基酸序列组合中一种：
1)QSVSSNL、GAS、QQYHSWPPLT；
2)QSVSSTY、GAS、QQRSNWLT；
3)QTIDNY、AAS、QQSYSTPPMYT；
4)QSISSY、SAS、QQSFTTPFT；
5)QSISSY、AAS、QQYSSYPHT；
所述H-CDR1、H-CDR2和H-CDR选自以下氨基酸序列组合中一种：
1)GYTFTSYG、ISAYNGNT、ARDFQGIAVADLDY；
2)GYTFTSYY、INPSGGST、ARGVGSYAMDV；
3)GFKFDDYP、ISWDGKTT、AKDRSRTGWGYGGMDV；
4)GDSVSSNSAA、TYYRSKWYN、ARSVGIARWDV；
5)GDSVSSNSAA、TYYRSKWYN、ARATGTASGWFDP。
2.根据权利要求1所述的双特异性抗体，其特征在于，所述CD22轻链包含如SEQ ID NO:1或SEQ ID NO:66所示的氨基酸序列或其功能性变体；所述CD22重链包含如SEQ ID NO:2或SEQ ID NO:67所示的氨基酸序列或其功能性变体。
3.根据权利要求1或2所述的双特异性抗体，其特征在于，所述CD19轻链包含如SEQ IDNO:3或SEQ ID NO:68所示的氨基酸序列或其功能性变体；所述CD19重链包含如SEQ ID NO:4或SEQ ID NO:69所示的氨基酸序列或其功能性变体。
4.根据权利要求3所述的双特异性抗体，其特征在于，所述双特异性抗体包括以下几种结构中的一种：
(1)CD22VL-linker4-CD22VH-Linker1-CD19VL-linker4-CD19VH；
(2)CD19VL-linker4-CD19VH-Linker1-CD22VL-linker4-CD22VH；
(3)CD22VL-linker4-CD22VH-Linker2-CD19VL-linker4-CD19VH；
(4)CD22VL-linker3-CD19VL-linker4-CD19VH-linker3-CD22VH；
(5)CD22VH-linker3-CD19VL-linker4-CD19VH-linker3-CD22VL。
5.根据权利要求4所述的双特异性抗体，其特征在于，所述linker4包含如SEQ ID NO:5所示的氨基酸序列或其功能性变体；和/或所述linker1包含如SEQ ID NO:6所示的氨基酸序列或其功能性变体；和/或所述linker2包含如SEQ ID NO:7所示的氨基酸序列或其功能性变体；和/或所述linker3包含如SEQ ID NO:8所示的氨基酸序列或其功能性变体。
6.根据权利要求1所述的双特异性抗体，其特征在于，所述双特异性抗体包含如SEQ IDNO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:75或SEQ IDNO:76所示的氨基酸序列或其功能性变体。
7.一种靶向CD22和CD19的嵌合抗原受体，其特征在于，包括以下几种结构中的一种：
(1)所述嵌合抗原受体依次连接胞外结构域、铰链区、跨膜区、胞内信号域，所述胞外结构域包含权利要求5所述的双特异性抗体，结构为以下结构中的一种：
(a)CD22VL-linker4-CD22VH-Linker1-CD19VL-linker4-CD19VH-8h-8TM-BBZ；或
(b)CD19VL-linker4-CD19VH-Linker1-CD22VL-linker4-CD22VH-8h-8TM-BBZ；或
(c)CD22VL-linker4-CD22VH-Linker2-CD19VL-linker4-CD19VH-8h-8TM-BBZ；或
(d)CD22VL-linker3-CD19VL-linker4-CD19VH-linker3-CD22VH-8h-8TM-BBZ；或
(e)CD22VH-linker3-CD19VL-linker4-CD19VH-linker3-CD22VL-8h-8TM-BBZ；或
(2)CD19VL-linker4-CD19VH-8h-8TM-BBZ-2A-CD22VL-linker4-CD22VH-8h`-28TM-28Z`；所述结构(2)中，CD19VL包含如SEQ ID NO:3所示的氨基酸序列或其功能性变体，CD19VH包含如SEQ ID NO:4所示的氨基酸序列或其功能性变体，CD22VL包含如SEQ ID NO:1所示的氨基酸序列或其功能性变体，CD22VH包含如SEQ ID NO:3所示的氨基酸序列或其功能性变体，linker4包含如SEQ ID NO:5所示的氨基酸序列或其功能性变体。
8.根据权利要求7所述的嵌合抗原受体，其特征在于，所述结构(2)中，所述2A包含如SEQ ID NO:14、SEQ ID NO:60或SEQ ID NO:61所示的氨基酸序列或其功能性变体。
9.根据权利要求7所述的嵌合抗原受体，其特征在于，所述8h包含如SEQ ID NO:15所示的氨基酸序列或其功能性变体；所述8h`包含如SEQ ID NO:16所示的氨基酸序列或其功能性变体。
10.根据权利要求7所述的嵌合抗原受体，其特征在于，所述8TM包含如SEQ ID NO:17所示的氨基酸序列或其功能性变体；所述28TM包含如SEQ ID NO:18所示的氨基酸序列或其功能性变体。
11.根据权利要求7所述的嵌合抗原受体，其特征在于，所述BBZ中Z的包含如SEQ IDNO:19所示的氨基酸序列或其功能性变体，所述BBZ中的BB包含如SEQ ID NO:20所示的氨基酸序列或其功能性变体；所述28Z`中的Z`包含如SEQ ID NO:21所示的氨基酸序列或其功能性变体，所述28Z`中的28包含如SEQ ID NO:22所示的氨基酸序列或其功能性变体。
12.根据权利要求7所述的嵌合抗原受体，其特征在于，所述嵌合抗原受体包含如SEQID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQID NO:77、SEQ ID NO:78、SEQ ID NO:83或SEQ ID NO:84所示的氨基酸序列或其功能性变体。
13.根据权利要求7-11任一所述的嵌合抗原受体，其特征在于，所述结构(1)中，嵌合抗原受体连接信号肽，所述信号肽包含如SEQ ID NO:23所示的氨基酸序列或其功能性变体；或所述结构(2)中，由2A连接的两组嵌合抗原受体中至少一组连接信号肽，所述信号肽包含如SEQ ID NO:23所示的氨基酸序列或其功能性变体。
14.核酸序列，其特征在于，编码权利要求7-13任一所述的嵌合抗原受体，其核酸序列包含如sequence NO.30、sequence NO.31、sequence NO.32、sequence NO.33、sequenceNO.34、sequence NO.35、sequence NO.81、sequence NO.82、SEQ ID NO:85或SEQ ID NO:86所示的序列。
15.根据权利要求14所述的核酸序列，其特征在于，所述核酸序列中还包括启动子，所述启动子选自巨细胞病毒立即早期基因启动子(CMV)、延伸因子1α启动子(EF1-α)、磷酸甘油酸激酶-1启动子(PGK)、泛素-C启动子(UBQ-C)、巨细胞病毒增强子/鸡β-肌动蛋白启动子(CAG)、多瘤病毒增强子/单纯疱疹胸苷激酶启动子(MC1)、β肌动蛋白启动子(β-ACT)、猿猴病毒40启动子(SV40)、骨髓增生肉瘤病毒增强子、阴性对照区缺失的dl587rev引物结合位点取代的(MND)启动子、缺氧启动子中一种。
16.根据权利要求15所述的核酸序列，其特征在于，缺氧启动子的序列如SEQ ID NO:62所示。
17.包含权利要求16所述的核酸序列的表达载体。
18.根据权利要求17所述的表达载体，其特征在于，所述表达载体选自慢病毒表达载体、逆转录病毒表达载体、腺病毒表达载体、腺相关病毒表达载体、DNA载体，RNA载体、质粒中的任一种。
19.一种工程化的细胞，其特征在于，所述细胞中转导有权利要求14-16所述的核酸序列或权利要求17-18任一所述的表达载体；优选地，所述细胞为T细胞、T细胞前体或NK细胞。
20.一种细胞制品，其特征在于，所述细胞制品包括权利要求19所述的工程化的细胞。
21.一种药物组合物，其特征在于，所述药物组合物包括权利要求1-6任一所述的双特异性抗体或权利要求7-13任一所述的嵌合抗原受体或权利要求14-16所述的核酸序列或权利要求17-18任一所述的表达载体或权利要求19所述的工程化的细胞或权利要求20所述的细胞制品。
22.权利要求1-6任一所述的双特异性抗体或权利要求7-13任一所述的嵌合抗原受体或权利要求14-16所述的核酸序列或权利要求17-18任一所述的表达载体或权利要求19所述的工程化的细胞或权利要求20所述的细胞制品或权利要求21所述的药物组合物在制备抗肿瘤药物中的应用。
23.根据权利要求22所述的应用，其特征在于，所述抗肿瘤药物为抗急性淋巴样白血病药物、抗慢性淋巴细胞白血病药物、抗慢性髓性白血病药物、抗非霍奇金淋巴瘤药物、抗霍奇金淋巴瘤药物、抗前列腺癌药物、抗结直肠癌药物、抗乳腺癌药物、抗卵巢癌药物、抗宫颈癌药物、抗胰腺癌药物、抗肺癌药物、抗肾癌药物、抗肝癌药物、抗脑癌药物或抗皮肤癌药物。
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