CN111032696B - Dctn1蛋白质与ret蛋白质的融合蛋白 - Google Patents

Dctn1蛋白质与ret蛋白质的融合蛋白 Download PDF

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CN111032696B
CN111032696B CN201880054583.2A CN201880054583A CN111032696B CN 111032696 B CN111032696 B CN 111032696B CN 201880054583 A CN201880054583 A CN 201880054583A CN 111032696 B CN111032696 B CN 111032696B
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林晃平
石田圭司
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Taiho Pharmaceutical Co Ltd
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Abstract

提供了一种其中DCTN1蛋白质的一部分与RET蛋白的一部分融合的新的多肽；编码多肽的多核苷酸；用于检测多核苷酸或多肽的方法；筛选抑制多核苷酸的表达或者多肽的表达和/或活性的化合物的方法；和含有抑制RET的化合物作为活性成分的药物组合物。

Description

DCTN1蛋白质与RET蛋白质的融合蛋白
发明领域
本发明涉及作为DCTN1蛋白质和RET蛋白质的融合蛋白的多肽；编码多肽的多核苷酸；检测多肽或多核苷酸的方法；靶向多肽或多核苷酸的化合物；和筛选化合物的方法。
背景技术
在日本，癌症是导致疾病死亡的主要原因，因此必须改善其治疗方法。尽管受甲状腺癌影响的人数正在增加，但由于大多数情况下该病进展缓慢，因此通过在初始阶段进行适当的治疗，表现出高的生存率。但是，这种疾病几乎没有主观症状，因此早期诊断对于适当的治疗来说是必不可少的。
甲状腺癌按组织学类型分为乳突癌、滤泡癌、髓样癌、未分化癌和恶性淋巴瘤。乳突癌约占甲状腺癌的80％，并且，未分化癌的发生率较低，但已知其预后很差(非专利文献1)。
已知乳突癌的发展主要是由于癌基因的激活，并且已经揭示发生相互排除的遗传异常，例如BRAF突变基因(50-60％)、RAS突变基因(10-20％)和RET融合基因(5-10％)。研究还报告了在非小细胞肺癌中，RET融合基因与其他驱动突变基因(例如EGFR突变基因)相互排斥地以1-2％的频率存在(非专利文献2-5)。
药物治疗是晚期甲状腺癌治疗的主要方式，并且已经批准了许多种多激酶抑制剂。但是，表现出对于驱动突变基因具有特异性的效果的药物仍未获批准。一项研究报告了RET融合基因阳性的肺癌患者表现出获益于抑制RET(非专利文献6)；在甲状腺癌中，鉴别基因异常(例如突变基因或融合基因)也是必要的，这可以作为药物效果的指标。
对于鉴别可能导致癌症的突变基因(突变蛋白)、融合基因(融合蛋白)等存在有强烈的需求；这是因为这种鉴别将阐明癌症的性质，并极大地有助于开发靶向这些突变基因或融合基因的新的癌症治疗药物或测试方法。但是，尚未完全阐明可能驱动癌症发展的突变基因、融合基因等，并且，鉴别与药物的治疗效果有关的基因异常将具有重大的意义。
引用列表
非专利文献
非专利文献1：Cancer,115(16),pp.3801-7(2009)
非专利文献2：Oncogene,22(29),pp.4578-80(2003)
非专利文献3：Cell,159(3),pp.676-90(2014)
非专利文献4：Cancer Discov.,3(6),pp.630-5(2013)
非专利文献5：Nature,511(7511),pp.543-50(2014)
非专利文献6：Lancet Respir Med.,5(1),pp.42-50(2017)
发明内容
技术问题
本发明的一个目的是提供一种新的多肽，该多肽是包括至少一部分RET蛋白质的融合蛋白；编码多肽的多核苷酸；检测多肽或多核苷酸的方法；靶向多肽或多核苷酸的化合物；和筛选化合物的方法。
解决问题的手段
本发明人进行了广泛的研究以实现该目的，并且在源自甲状腺癌患者的细胞中鉴别出其中DCTN1蛋白质的一部分与RET蛋白质的一部分融合的新型多肽，以及编码该多肽的多核苷酸。发明人还发现了用于在癌细胞中检测本发明的多核苷酸或多肽的方法，以及筛选抑制多核苷酸的表达或者多肽的表达和/或活性的化合物的方法。这是一项新的发现，不能从现有技术中预测，在各种各样的蛋白质当中，含有DCTN1蛋白质的N-端部分和RET蛋白质的C-端部分的组合的融合蛋白天然地在细胞内发生；而且，由于DCTN1和RET的融合基因起到癌症驱动因子(cancer driver)的作用，因此融合蛋白可用于癌症的诊断。他们还发现了一种药物组合物，其含有抑制RET的化合物作为活性成分，用于治疗具有多肽和/或多核苷酸的表达的癌症患者，从而完成了本发明。
具体地，本发明提供以下主题。
第1项.
一种多肽，其中DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合。
第2项.
根据第1项所述的多肽，其选自以下(a)至(c)：
(a)多肽，其包括SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ IDNO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ IDNO：22或SEQ ID NO：24所示的氨基酸序列；
(b)多肽，其包括在SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQID NO：22或SEQ ID NO：24所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列；和
(c)多肽，其包括与SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQID NO：22或SEQ ID NO：24所示的氨基酸序列具有至少90％同一性的氨基酸序列。
第3项.
一种多核苷酸，其编码根据第1或2项所述的多肽。
第4项.
根据第3项所述的多核苷酸，其选自以下(d)至(f)：
(d)多核苷酸，其编码包括SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ ID NO：22或SEQ ID NO：24所示的氨基酸序列的多肽；
(e)多核苷酸，其编码包括在SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ IDNO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ IDNO：20、SEQ ID NO：22或SEQ ID NO：24所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列的多肽；和
(f)多核苷酸，其编码包括与SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ IDNO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ IDNO：20、SEQ ID NO：22或SEQ ID NO：24所示的氨基酸序列具有至少90％同一性的氨基酸序列的多肽。
第5项.
根据第3项所述的多核苷酸，其选自以下(g)至(i)：
(g)多核苷酸，其包括SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQID NO：9、SEQ ID NO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQID NO：21或SEQ ID NO：23所示的碱基序列；
(h)多核苷酸，其在严格条件下与包括与SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQ ID NO：9、SEQ ID NO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQ ID NO：21或SEQ ID NO：23所示的碱基序列互补的碱基序列的多核苷酸杂交；和
(i)多核苷酸，其与SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQID NO：9、SEQ ID NO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQID NO：21或SEQ ID NO：23所示的碱基序列具有至少90％同一性。
第6项.
一种表达载体，其包括根据第3-5项中任一项所述的多核苷酸。
第7项.
一种用根据第3-5项中任一项所述的多核苷酸转染的细胞。
第8项.
一种抗体，其特异性结合根据第1或2项所述的多肽。
第9项.
一种用于检测样品中根据第1或2项所述的多肽的存在的方法。
第10项.
一种引物或探针，其用于检测样品中根据第3-5项中任一项所述的多核苷酸的存在，该引物或探针是选自以下(j)至(l)的多核苷酸：
(j)多核苷酸，其是选自与编码DCTN1蛋白质的多核苷酸杂交的探针和与编码RET蛋白质的多核苷酸杂交的探针的至少一种探针；
(k)多核苷酸，其是与编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的融合点杂交的探针；和
(1)多核苷酸，其是一组设计成将融合点夹在编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的有义引物和反义引物。
第11项.
一种检测样品中根据第3-5项中任一项所述的多核苷酸的存在的方法。
第12项.
一种诊断患者患有癌症的方法，通过根据第9或11项所述的检测方法在来源于患者的样品中检测到根据第1或2项所述的多肽或者根据第3-5项中任一项所述的多核苷酸的存在时，诊断患者患有癌症。
第13项.
一种用于治疗癌症的药物组合物，所述癌症对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性，该组合物包括抑制RET的化合物作为活性成分。
第14项.
一种筛选化合物的方法，所述化合物抑制根据第1或2项所述的多肽的表达和/或活性或者根据第3-5项中任一项所述的多核苷酸的表达，该方法包括以下步骤(1)和(2)：
(1)使根据第1或2项所述的多肽、表达根据第1或2项所述的多肽或者根据第3-5项中任一项所述的多核苷酸的细胞、或者根据第7项所述的细胞与测试化合物接触的步骤；和
(2)测量步骤(1)中根据第1或2项所述的多肽的表达和/或活性或者根据第3-5项中任一项所述的多核苷酸的表达是否受到抑制的步骤，或者测量步骤(1)中细胞的生长是否受到抑制的步骤。
第15项.
一种使用根据第1或2项所述的多肽或者根据第3-5项中任一项所述的多核苷酸作为确定使用抑制RET的化合物的化学疗法是否有效的指标的方法，
该方法包括：当通过根据第9项所述的检测方法在样品中检测到根据第1或2项所述的多肽时，和/或当通过根据第11项所述的检测方法在样品中检测到根据第3-5项中任一项所述的多核苷酸的存在时，确定使用抑制RET的化合物的化学疗法是有效的。
第16项.
一种用于检测癌症的生物标志物，该生物标志物包括选自其中DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合的多肽和编码所述多肽的多核苷酸的至少一种。
第17项.
一种治疗癌症的方法，该方法包括
向对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症患者给予使用抑制RET的化合物的化学疗法。
第18项.
一种治疗癌症的方法，该方法包括
检测来源于测试受试者的样品中根据第1或2项所述的多肽的存在和/或根据第3-5项中任一项所述的多核苷酸的存在，和
当检测到根据第1或2项所述的多肽的存在和/或检测到根据第3-5项中任一项所述的多核苷酸的存在时，对测试受试者给予使用抑制RET的化合物的化学疗法。
第19项.
一种抑制RET的化合物，用于治疗癌症患者，所述患者对于DCTN1基因和RET基因的融合基因呈阳性，和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性。
第20项.
抑制RET的化合物在制造用于癌症治疗的药物组合物中的用途，所述药物组合物用于治疗对于DCTN1基因和RET基因的融合基因呈阳性、和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症患者。
第21项.
一种用于制造试剂的方法，该试剂用于使用检测样品中根据第1或2项所述的多肽的存在的手段，和/或检测样品中根据第3-5项中任一项所述的多核苷酸的存在的手段，确定使用抑制RET的化合物的化学疗法是否有效。
第22项.
一种抗DCTN1抗体和抗RET抗体的组合，用于检测根据第3-5项中任一项所述的多核苷酸的存在。
第23项.
根据第8项所述的抗体、根据第22项所述的抗体的组合、或者根据第10项所述的引物或探针在制造检测试剂中的用途，所述检测试剂用于检测根据第1或2项所述的多肽的存在、或者根据第3-5项中任一项所述的多核苷酸的存在。
发明的有益效果
本发明阐明了本发明的多核苷酸和/或多肽在癌细胞中是特异性表达的。本发明的多核苷酸、多肽和表达多核苷酸和/或多肽的细胞可以用于筛选抑制本发明的多核苷酸的表达或多肽的表达和/或活性的化合物的方法。利用本发明的多核苷酸和/或多肽的存在作为指标，能够检测对于DCTN1基因和RET基因的融合基因呈阳性的靶标、和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的靶标。抑制RET的化合物可用作治疗癌症的药物组合物，上述癌症对于DCTN1基因和RET基因的融合基因呈阳性，和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性
附图说明
图1：使用Droplet Digital PCR(ddPCR)确认DCTN1-RET融合基因和GAPDH在甲状腺癌组织中的表达。
图2：使用Droplet Digital PCR(ddPCR)确认DCTN1-RET融合基因和GAPDH在正常甲状腺组织中的表达。
图3：确认全长DCTN1-RET融合基因在正常甲状腺组织和甲状腺癌组织中的表达。
图4：确认DCTN1-RET融合蛋白在表达DCTN1-RET融合基因的NIH/3T3细胞中的表达：
a)使用抗磷酸化RET抗体检测DCTN1-RET融合蛋白；
b)使用抗RET抗体检测DCTN1-RET融合蛋白；和
c)使用抗DCTN1抗体检测DCTN1-RET融合蛋白。
图5：确认表达DCTN1-RET融合基因的NIH/3T3细胞在三维培养基中的生长。
N＝3，平均值+SD。
图6：在体内确认表达DCTN1-RET融合基因的NIH/3T3细胞的致瘤性。
图7：在表达DCTN1-RET融合基因的NIH/3T3细胞中确认磷酸化RET的表达被RETsiRNA抑制。
图8：确认RET siRNA对表达DCTN1-RET融合基因的NIH/3T3细胞的生长抑制效果。
图9：确认抑制RET的化合物对表达DCTN1-RET融合基因的NIH/3T3细胞中的磷酸化RET的表达的抑制。
具体实施方式
本发明涉及新的多核苷酸或多肽；用于检测多核苷酸或多肽的方法；靶向多核苷酸或多肽的化合物；和筛选化合物的方法。
本发明提供一种多肽，其中DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合(其在下文中可以称为“本发明的多肽”)。本发明还提供编码该多肽的多核苷酸(其在下文中可以称为“本发明的多核苷酸”)。
本发明中的“DCTN1(动力蛋白激活蛋白(Dynactin)亚基1)蛋白质”也被称为与150kDa动力蛋白相关的多肽蛋白质或DAP-150蛋白质，并包括人或非人哺乳动物的DCTN1蛋白质，优选人DCTN1蛋白质。DCTN1蛋白质在人体内由位于2p13.1上的基因编码。在本发明中，“DCTN1蛋白质”包括同种型(其剪接变体)，并且人源DCTN1蛋白质的实例包括了包括GenPept登录号NP_004073、NP_075408、NP_001128512、NP_001128513、NP_001177765或NP_001177766所示的氨基酸序列的多肽。更具体地，实例包括了包括SEQ ID NO：25、SEQ IDNO：26、SEQ ID NO：27、SEQ ID NO：28、SEQ ID NO：29或SEQ ID NO：30所示的氨基酸序列的多肽。另外，在本发明中“DCTN1蛋白质的N-端部分”是指含有DCTN1蛋白质的N-端侧的部分或整个卷曲螺旋(coiled-coil)结构域的多肽，优选地，是指含有DCTN1蛋白质N-端侧的整个卷曲螺旋结构域的多肽。
在本发明中“RET蛋白质”也称为RET原癌基因蛋白质、RET受体酪氨酸激酶蛋白质或转染过程中重排的蛋白质；其包括人或非人哺乳动物的RET蛋白质，优选人RET蛋白质。RET蛋白质在人体内由位于10q11.2上的基因编码。在本发明中，“RET蛋白质”包括同种型(其剪接变体)，并且人源RET蛋白质的实例包括了包括GenPept登录号NP_066124或NP_065681所示的氨基酸序列的多肽。更具体地，实例包括了包括SEQ ID NO：31或SEQ ID NO：32所示的氨基酸序列的多肽。另外，在本发明中“RET蛋白质的C-端部分”是指含有RET蛋白质的C-端侧的激酶结构域的多肽。
在本发明中，“DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合的多肽”是其中含有DCTN1蛋白质的N-端侧的部分或整个卷曲螺旋结构域的多肽与含有RET蛋白质的C-端侧的激酶结构域的多肽融合的多肽，优选为其中含有DCTN1蛋白质的N-端侧的整个卷曲螺旋结构域的多肽与含有RET蛋白质的C-端侧的激酶结构域的多肽融合的多肽，更优选为选自以下(a)至(c)的多肽。这些多肽优选具有激酶活性和/或细胞增殖作用。
(a)多肽，其包括SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ IDNO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ IDNO：22或SEQ ID NO：24所示的氨基酸序列；
(b)多肽，其包括在SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQID NO：22或SEQ ID NO：24所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列；和
(c)多肽，其包括与SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQID NO：22或SEQ ID NO：24所示的氨基酸序列具有至少90％同一性的氨基酸序列。
更优选地，多肽选自以下(a)至(c)。这些多肽优选具有激酶活性或细胞增殖作用。
(a)多肽，其包括SEQ ID NO：18所示的氨基酸序列。
(b)多肽，其包括在SEQ ID NO：18所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列。
(c)多肽，其包括与SEQ ID NO：18所示的氨基酸序列具有至少90％同一性的氨基酸序列。
在本发明中“DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合的多肽”包括了包括在SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ ID NO：10、SEQ IDNO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ ID NO：22或SEQID NO：24所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列的多肽(上文(b)项)。包括DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合的这类氨基酸序列的多肽的实例包括了包括SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQID NO：22或SEQ ID NO：24所示的氨基酸序列的多肽(其中DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合)的同种型。这些多肽优选具有激酶活性或细胞增殖作用。如本文所用，“取代、缺失或添加的几个氨基酸”是指，例如，优选1-10个氨基酸，更优选1-5个氨基酸。“添加”包括在N-端或C-端添加一个至几个氨基酸，或者在两个端均添加一个至几个氨基酸。
一个或几个氨基酸被取代的多肽的实例包括其中在作为包括GenPept登录号NP_066124(SEQ ID NO：31)或NP_065681(SEQ ID NO：32)所示氨基酸序列的RET蛋白质的关守位点(gatekeeper site)的804位(SEQ ID NO：2和SEQ ID NO：4中的1325位、SEQ ID NO：6和SEQ ID NO：8中的1191位、SEQ ID NO：10和SEQ ID NO：12中的1300位、SEQ ID NO：14和SEQID NO：16中的1186位、SEQ ID NO：18和SEQ ID NO：20中的1283位、SEQ ID NO：22和SEQ IDNO：24中的1318位)上的缬氨酸在被亮氨酸、蛋氨酸或谷氨酸取代的多肽；以及其中在806位(SEQ ID NO：2和SEQ ID NO：4中的1327位、SEQ ID NO：6和SEQ ID NO：8中的1193位、SEQ IDNO：10和SEQ ID NO：12中的1302位、SEQ ID NO：14和SEQ ID NO：16中的1188位、SEQ ID NO：18和SEQ ID NO：20中的1285位、SEQ ID NO：22和SEQ ID NO：24中的1320位)上的酪氨酸被半胱氨酸、谷氨酸、丝氨酸、组氨酸或天冬酰胺取代的多肽。
实例还包括关守位点以外的位置处的氨基酸，但不限于其中在768位(SEQ ID NO：2和SEQ ID NO：4中的1289位、SEQ ID NO：6和SEQ ID NO：8中的1155位、SEQ ID NO：10和SEQID NO：12中的1264位、SEQ ID NO：14和SEQ ID NO：16中的1150位、SEQ ID NO：18和SEQ IDNO：20中的1247位、SEQ ID NO：22和SEQ ID NO：24中的1282位)上的谷氨酸被天冬氨酸取代的多肽；其中在883位(SEQ ID NO：2和SEQ ID NO：4中的1404位、SEQ ID NO：6和SEQ ID NO：8中的1270位、SEQ ID NO：10和SEQ ID NO：12中的1379位、SEQ ID NO：14和SEQ ID NO：16中的1265位、SEQ ID NO：18和SEQ ID NO：20中的1362位、SEQ ID NO：22和SEQ ID NO：24中的1397位)上的丙氨酸被苯丙氨酸或丝氨酸取代的多肽；其中在884位(SEQ ID NO：2和SEQ IDNO：4中的1405位、SEQ ID NO：6和SEQ ID NO：8中的1271位、SEQ ID NO：10和SEQ ID NO：12中的1380位、SEQ ID NO：14和SEQ ID NO：16中的1266位、SEQ ID NO：18和SEQ ID NO：20中的1363位、SEQ ID NO：22和SEQ ID NO：24中的1398位)上的谷氨酸被缬氨酸取代的多肽；其中在891位(SEQ ID NO：2和SEQ ID NO：4中的1412位、SEQ ID NO：6和SEQ ID NO：8中的1278位、SEQ ID NO：10和SEQ ID NO：12中的1387位、SEQ ID NO：14和SEQ ID NO：16中的1273位、SEQ ID NO：18和SEQ ID NO：20中的1370位、SEQ ID NO：22和SEQ ID NO：24中的1405位)上的丝氨酸被丙氨酸或亮氨酸取代的多肽；以及其中在918位(SEQ ID NO：2和SEQ ID NO：4中的1439位、SEQ ID NO：6和SEQ ID NO：8中的1305位、SEQ ID NO：10和SEQ ID NO：12中的1414位、SEQ ID NO：14和SEQ ID NO：16中的1300位、SEQ ID NO：18和SEQ ID NO：20中的1397位、SEQ ID NO：22和SEQ ID NO：24中的1432位)上的蛋氨酸被苏氨酸取代的多肽。
本发明的DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合的多肽包括了包括与SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ ID NO：22或SEQ IDNO：24所示的氨基酸序列在其适当比对时具有至少90％同一性的氨基酸序列的多肽(上文(c)项)。这些多肽优选具有激酶活性或细胞增殖作用。
与SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ ID NO：10、SEQ IDNO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ ID NO：22或SEQID NO：24所示的氨基酸序列的同一性优选为至少90％，更优选为至少95％，再更优选为至少98％。氨基酸序列的同一性可以通过常用方法来计算。
除构成本发明的多肽的氨基酸序列以外，本发明的多肽还可以包括构成蛋白质标记的氨基酸。可用的标记的实例包括本领域技术人员公知的那些标记；例如，可用的标记包括用于提高表达效率的标记和用于提高提纯效率的标记，例如His标记、Myc标记和FLAG标记。
本发明的多核苷酸编码其中DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合的多肽，并且优选为选自以下(d)至(i)的多核苷酸。这些多核苷酸优选编码具有激酶活性或细胞增殖作用的多肽。
(d)多核苷酸，其编码包括SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ ID NO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ ID NO：22或SEQ ID NO：24所示的氨基酸序列的多肽。
(e)多核苷酸，其编码包括在SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ IDNO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ IDNO：20、SEQ ID NO：22或SEQ ID NO：24所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列的多肽。
(f)多核苷酸，其编码包括与SEQ ID NO：2、SEQ ID NO：4、SEQ ID NO：6、SEQ IDNO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ ID NO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ IDNO：20、SEQ ID NO：22或SEQ ID NO：24所示的氨基酸序列具有至少90％同一性的氨基酸序列的多肽。
(g)多核苷酸，其包括SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQID NO：9、SEQ ID NO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQID NO：21或SEQ ID NO：23所示的碱基序列。
(h)多核苷酸，其在严格条件下与包括与SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQ ID NO：9、SEQ ID NO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQ ID NO：21或SEQ ID NO：23所示的碱基序列互补的碱基序列的多核苷酸杂交。
(i)多核苷酸，其与SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQID NO：9、SEQ ID NO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQID NO：21或SEQ ID NO：23所示的碱基序列具有至少90％同一性。
更优选地，本发明的多核苷酸选自以下(d)至(i)。这些多核苷酸优选编码具有激酶活性或细胞增殖作用的多肽。
(d)多核苷酸，其编码包括SEQ ID NO：18所示的氨基酸序列的多肽。
(e)多核苷酸，其编码包括在SEQ ID NO：18所示的氨基酸序列中取代、缺失或添加一个或几个氨基酸的氨基酸序列的多肽。
(f)多核苷酸，其编码包括与SEQ ID NO：18所示的氨基酸序列具有至少90％同一性的氨基酸序列的多肽。
(g)多核苷酸，其包括SEQ ID NO：17所示的碱基序列。
(h)多核苷酸，其在严格条件下与包括与SEQ ID NO：17所示的碱基序列互补的碱基序列的多核苷酸杂交。
(i)多核苷酸，其与SEQ ID NO：17所示的碱基序列具有至少90％同一性。
本发明的多核苷酸不仅包括其双链DNA，而且还包括构成双链DNA的各种类型的单链DNA和RNA，例如有义链和反义链。反义链可以用作探针等。DNA包括通过克隆、化学合成或其组合获得的DNA，例如cDNA和基因组DNA。另外，除编码本发明的多肽的碱基序列以外，本发明的多核苷酸还可以添加有碱基序列，例如非翻译区(UTR)序列。
如本文所用，严格条件包括例如在《分子克隆：实验室手册(Molecular Cloning:ALaboratory Manual)》(第二版，J.Sambrook等，1989)中描述的条件。具体地，严格条件包括使得含有6×SSC(1×SSC组成：0.15M氯化钠、0.015M柠檬酸钠、pH 7.0)、0.5％SDS、5×Denhalt和100mg/mL鲱鱼精DNA的溶液与探针一起在65℃下等温处理8-16小时以进行杂交的条件。
与SEQ ID NO：1、SEQ ID NO：3、SEQ ID NO：5、SEQ ID NO：7、SEQ ID NO：9、SEQ IDNO：11、SEQ ID NO：13、SEQ ID NO：15、SEQ ID NO：17、SEQ ID NO：19、SEQ ID NO：21或SEQID NO：23所示的碱基序列的同一性优选为至少90％，更优选为至少95％，再更优选为至少98％。碱基序列的同一性可以通过常用方法来计算。
在本说明书中，“具有激酶活性或细胞增殖作用”中的术语“具有激酶活性”是指具有使酪氨酸磷酸化的酶活性。另外，“具有激酶活性或细胞增殖作用”中的术语“具有细胞增殖作用”是指，与未转染本发明的多核苷酸和/或多肽的细胞相比，将本发明的多核苷酸和/或多肽转染到细胞中提高了细胞增殖能力。例如，可以如下确认该作用：将多核苷酸和/或多肽转染到依赖于细胞因子增殖的细胞系中，如果细胞系不依赖于细胞因子而增殖，则多核苷酸和/或多肽具有细胞增殖作用。
例如，使用保留DCTN1基因和RET基因的融合基因的从甲状腺癌等制备的cDNA文库或基因组DNA文库，使用与本发明的多核苷酸的部分碱基序列特异性杂交的引物，可以提取本发明的多核苷酸。对于该引物，可以使用任何序列和任何长度的任何引物，只要该引物与本发明的多核苷酸的至少一部分或其反义链特异性杂交即可。也可以使用人工合成多核苷酸的方法(Nat.Methods,11:499-507,2014)。
本发明的表达载体没有特别限定，只要表达载体包括本发明的多核苷酸、允许表达本发明的多肽即可。实例包括通过将本发明的多核苷酸插入根据使用宿主适当选择的已知表达载体而获得的表达载体。
对宿主没有特别限定，只要宿主是可以经历转化的活细胞即可，实例包括细菌，例如大肠杆菌(E.coli)和枯草芽孢杆菌(Bacillus subtilis)；真菌，例如酵母和丝状真菌；昆虫细胞，例如Sf9细胞；昆虫，例如蚕；动物细胞；和植物或植物来源的细胞。
用于插入本发明的多核苷酸的载体没有特别限定，只要载体在宿主中可复制即可。载体可以根据例如引入宿主的类型和引入方法而适当选择。实例包括质粒DNA、噬菌体DNA和病毒载体。对于用于构建表达载体的载体DNA，可以使用广泛流行且容易获得的载体DNA。实例包括pUC19(Takara Bio Inc.)、pTV118N(Takara Bio Inc.)、pMAMneo(ClontechLaboratories，Inc.)、pGEX(GE Healthcare)、pET160(Invitrogen)、pDEST(Invitrogen)、pIEx(Merck Millipore)和pBacPAK(Clontech Laboratories，Inc.)。病毒载体的实例包括DNA病毒和RNA病毒，例如杆状病毒载体、逆转录病毒载体、慢病毒载体(例如人免疫缺陷病毒或HIV)、腺病毒载体、腺相关病毒载体(AAV载体)、疱疹病毒、痘苗病毒、痘病毒、脊髓灰质炎病毒、信德比斯(Sindbis)病毒、仙台病毒和猿猴病毒40(SV-40)。
例如，通过原生质体方法、感受态细胞方法或电穿孔方法，可以进行使用表达载体的宿主的转化。获得的转化体可以在适当条件下在含有可以被宿主利用碳源、氮源、金属盐、维生素等的培养基中培养。
用根据本发明的多核苷酸转染的细胞的实例包括通过本发明的表达载体转化的细胞，以及已经通过基因组编辑引入本发明的多核苷酸的细胞。使用的细胞包括上文列出的宿主细胞。用于确认细胞是否已被表达载体转化的方法的实例包括：检测本发明的多肽的存在的方法，和检测本发明的多核苷酸的存在的方法。
“已经通过基因组编辑引入本发明的多核苷酸的细胞”优选为具有通过基因组编辑融合独立存在的DCTN1基因和RET基因而获得的基因的细胞，更优选为具有通过基因组编辑融合在分别独立存在的DCTN1基因和RET基因中DCTN1的外显子27和RET的外显子12而获得的基因的细胞。这些细胞可以通过常用的方法来制备，实例包括例如Cell Rep.,9(4),pp.1219-1227(2014)、Nat.Commun.,5,3728(2014)中描述的方法。用于确认细胞是否是通过基因组编辑导入本发明的多核苷酸的细胞的方法的实例包括：检测本发明的多肽的存在的方法，和检测本发明的多核苷酸的存在的方法。
本发明的多肽可以通过以下方法获得：通过在合适的条件下在适合于细胞培养的培养基中培养由本发明的表达载体转化的细胞，制备培养液和/或细胞；然后通过常规方法从培养液和/或细胞中收集并提纯蛋白质。本发明的多肽还可以通过以下方法获得：将含有本发明的多核苷酸的表达载体或编码本发明的多核苷酸的模板RNA或模板DNA并入无细胞蛋白质合成系统(例如人细胞系来源的细胞提取物、兔网织红细胞提取物、小麦胚芽提取物和大肠杆菌提取物)；在合适的条件下孵育产物；通过常规方法从获得的反应液中收集并提纯蛋白质。
在本发明中，与本发明的多肽特异性结合的抗体包括与DCTN1蛋白质的N-端部分和RET蛋白质的C-端部分的融合点特异性结合的抗体。抗体是指与DCTN1蛋白质的N-端部分和RET蛋白质的C-端部分的融合点特异性结合、但不结合野生型DCTN1并且不结合野生型RET蛋白质的抗体。
在本发明中，“DCTN1蛋白质的N-端部分和RET蛋白质的C-端部分的融合点”中的术语“融合点”是指源自DCTN1蛋白的N-端部分的多肽与源自RET蛋白质的C-端部分的多肽融合的点。SEQ ID NO：2中的融合点是具有SEQ ID NO：2的1-1233位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：2的1234-1635位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：4中的融合点是具有SEQ ID NO：4的1-1233位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：4的1234-1593位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：6中的融合点是具有SEQ ID NO：6的1-1099位氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：6的1100-1501位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：8中的融合点是具有SEQ ID NO：8的1-1099位氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：8的1100-1459位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：10中的融合点是具有SEQID NO：10的1-1208位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：10的1209-1610位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：12中的融合点是具有SEQ ID NO：12的1-1208位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：12的1209-1568位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ IDNO：14中的融合点是具有SEQ ID NO：14的1-1094位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：14的1095-1496位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：16中的融合点是具有SEQ ID NO：16的1-1094位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：16的1095-1454位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：18中的融合点是具有SEQ ID NO：18的1-1191位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：18的1192-1593位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：20中的融合点是具有SEQ ID NO：20的1-1191位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：20的1192-1551位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：22中的融合点是具有SEQID NO：22的1-1226位的氨基酸序列的多肽(源自DCTN1的N-端部分)与具有SEQ ID NO：22的1227-1628位的氨基酸序列的多肽(源自RET的C-端部分)融合的点。SEQ ID NO：24中的融合点是具有SEQ ID NO：24的1-1226位的氨基酸序列的多肽(源自DCTN1的N端部分)与具有SEQID NO：24的1227-1586位的氨基酸序列的多肽(源自RET的C端部分)融合的点。
抗体的实例包括免疫球蛋白(例如，IgA、IgD、IgE、IgG、IgM和IgY)、Fab片段、F(ab')2片段、单链抗体片段(scFv)、单结构域抗体和双抗体(Nat.Rev.Immunol.,6:343-357,2006)。这些包括，但不限于，例如人抗体、人源化抗体、嵌合抗体、小鼠抗体、美洲驼抗体和鸡抗体的单克隆抗体和多克隆抗体。
抗体可以通过各种已知方法制备，制备方法没有特别限定。已知方法包括以下方法：将本发明的多肽、含有DCTN1蛋白质的N-端部分和RET蛋白质的C-端部分的融合点的多肽片段等接种到免疫的动物中以激活动物的免疫系统，收集动物的血清以获得多克隆抗体；以及通过例如杂交瘤方法和噬菌体展示方法获得单克隆抗体的方法。
筛选抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物的方法可以通过包括以下步骤(1)和(2)的方法进行。
具体地，本发明的筛选方法通过包括以下步骤的方法进行：
(1)使本发明的多肽或表达本发明的多肽和/或多核苷酸的细胞与测试化合物接触的步骤；和
(2)测量步骤(1)中本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达是否受到抑制的步骤，或者测量步骤(1)中细胞的生长是否受到抑制的步骤。
更优选地，本发明的筛选方法是包括以下步骤(1)和(2)的方法。
(1)使表达本发明的多肽和/或多核苷酸的细胞与测试化合物接触的步骤。
(2)测量步骤(1)中的细胞生长是否受到抑制的步骤。
筛选抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物的方法可以通过包括以下步骤(1)至(3)的方法进行。
具体地，本发明的筛选方法通过包括以下步骤的方法进行：
(1)使本发明的多肽或者表达本发明的多肽和/或多核苷酸的细胞与测试化合物接触的步骤；
(2)测量步骤(1)中本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达是否受到抑制的步骤，或者测量步骤(1)中细胞的生长是否受到抑制的步骤；和
(3)当步骤(2)中本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达受到抑制时，或者当步骤(2)中步骤(1)中的细胞的生长受到抑制时，确定测试化合物抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的步骤。
更优选地，本发明的筛选方法是包括以下步骤(1)至(3)的方法。
(1)使表达本发明的多肽和/或多核苷酸的细胞与测试化合物接触的步骤。
(2)测量步骤(1)中的细胞的生长是否受到抑制的步骤。
(3)当步骤(2)中步骤(1)中的细胞的生长受到抑制时，确定测试化合物抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的步骤。
“表达本发明的多肽和/或多核苷酸的细胞”包括通过本发明的表达载体转化的细胞、通过基因组编辑引入本发明的多核苷酸的细胞、表达本发明的多肽和/或多核苷酸的原代培养细胞、表达本发明的多肽和/或多核苷酸的细胞系、和表达本发明的多肽和/或多核苷酸的癌症患者来源的细胞。确认细胞是否表达本发明的多肽和/或多核苷酸的方法的实例包括检测本发明的多肽的存在的方法和检测本发明的多核苷酸的存在的方法。
在本发明中，例如，术语“本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达受到抑制”中的“本发明的多肽的表达或本发明的多核苷酸的表达受到抑制”是指以下含义。使表达本发明的多肽和/或多核苷酸的细胞与测试化合物接触，使用检测本发明的多肽或多核苷酸的存在的方法来评估细胞中本发明的多肽或多核苷酸的表达水平。与没有接触过测试化合物的细胞相比，当与测试化合物接触过的细胞在统计学上显著降低了本发明的多肽或多核苷酸的表达水平时，确定本发明的多肽或多核苷酸的表达受到抑制。
例如，术语“本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达受到抑制”中的术语“本发明的多肽的活性受到抑制”是指以下含义。与没有接触过测试化合物的多肽或细胞相比，当与测试化合物接触过的本发明的多肽或表达本发明的多肽的细胞导致了统计学上显著降低的酪氨酸磷酸化百分比时，确定本发明的多肽的活性受到抑制。
与没有接触过测试化合物的细胞相比，当接触过测试化合物的表达本发明的多肽的细胞表现出统计学上显著抑制的细胞生长时，确定本发明的多肽的活性受到抑制。
在本发明中，“酪氨酸磷酸化”不仅包括RET蛋白质(包括与其他蛋白质融合的RET蛋白质)中的酪氨酸磷酸化，还包括RET信号传导下游的蛋白质中的酪氨酸磷酸化。RET信号传导下游的蛋白质的实例包括STAT、AKT和ERK。酪氨酸磷酸化优选为RET蛋白质(包括与其他蛋白质融合的RET蛋白质)中的酪氨酸磷酸化。
例如，通过蛋白质免疫印迹、免疫沉淀、免疫组织化学、ELISA或流式细胞术，可以使用磷酸化RET特异性抗体来测量“酪氨酸磷酸化百分比”。
在本发明中，“样品”不仅包括生物样品(例如细胞、组织、器官、体液(例如血液和淋巴液)、消化液和尿液)，还包括核酸提取物(例如基因组DNA提取物、mRNA提取物和由mRNA提取物制备的cDNA制剂和cRNA制剂)和从这些生物样品获得的蛋白质提取物。样品可以是经过福尔马林固定、酒精固定、冷冻处理或石蜡包埋的样品。使用的生物样品可以是从活体收集的那些，并且优选为源自癌症患者的样品，更优选为含有肿瘤细胞的样品。可以根据生物样品的类型适当地选择收集生物样品的方法。
本发明包括检测样品中本发明的多肽的存在的方法。
在本发明中，检测样品中本发明的多肽的存在的方法包括：根据常用方法例如ELISA、蛋白质免疫印迹或免疫组织化学染色的检测方法，其使用特异性结合本发明的多肽的抗体；和FRET(荧光共振能量转移)，其使用特异性结合DCTN1蛋白质的抗体和特异性结合RET蛋白质的抗体。检测方法优选为ELISA、蛋白质免疫印迹或免疫组织化学染色，其使用特异性结合本发明的多肽的抗体。
特异性结合DCTN1蛋白质的抗体和特异性结合RET蛋白质的抗体优选为与来自DCTN1蛋白质中的融合点的N-端部分结合的抗体，和与来自RET蛋白质中的融合点的C-端部分结合的抗体。这些抗体可以是市售产品，或者通过常规已知的方法制备。
在本发明中，检测样品中本发明的多肽的存在的方法优选地包括使用特异性结合本发明的多肽的抗体或者特异性结合DCTN1蛋白质的抗体和特异性结合RET蛋白质的抗体检测本发明的多肽的步骤；更优选地包括使用特异性结合本发明的多肽的抗体检测本发明的多肽的步骤。检测本发明的多肽的存在的手段没有特别限定，其实例包括特异性结合DCTN1蛋白质的抗体和特异性结合RET蛋白质的抗体的组合；和特异性结合本发明的多肽的抗体。
本发明包括用于检测样品中本发明的多核苷酸的存在的引物或探针。在本发明中，检测本发明的多肽的存在的手段没有特别限定，其实例包括用于检测本发明的多核苷酸的存在的引物或探针。
引物或探针包括选自以下(j)至(1)的多核苷酸：
(j)多核苷酸，其是选自与编码DCTN1蛋白质的多核苷酸杂交的探针和与编码RET蛋白质的多核苷酸杂交的探针的至少一种探针；
(k)多核苷酸，其是与编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的融合点杂交的探针；和
(1)多核苷酸，其是一组设计成将融合点夹在编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的有义引物和反义引物。
在本发明中，术语“编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的融合点”中的“融合点”是指编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸融合的点。SEQ ID NO：1中的融合点是具有SEQ ID NO：1的1-3699位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：1的3700-4905位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：3中的融合点是具有SEQ IDNO：3的1-3699位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ IDNO：3的3700-4779位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：5中的融合点是具有SEQ ID NO：5的1-3297位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：5的3298-4503位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：7中的融合点是具有SEQ ID NO：7的1-3297位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：7的3298-4377位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ IDNO：9中的融合点是具有SEQ ID NO：9的1-3624位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：9的3625-4830位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：11中的融合点是具有SEQ ID NO：11的1-3624位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：11的3625-4704位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：13中的融合点是具有SEQ ID NO：13的1-3282位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：13的3283-4488位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：15中的融合点是具有SEQ ID NO：15的1-3282位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：15的3283-4362位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：17中的融合点是具有SEQ ID NO：17的1-3573位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：17的3574-4779位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：19中的融合点是具有SEQ ID NO：19的1-3573位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：19的3574-4653位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：21中的融合点是具有SEQ ID NO：21的1-3678位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：21的3679-4884位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。SEQ ID NO：23中的融合点是具有SEQ ID NO：23的1-3678位的碱基序列的多核苷酸(源自编码DCTN1蛋白质的多核苷酸)与具有SEQ ID NO：23的3679-4758位的碱基序列的多核苷酸(源自编码RET蛋白质的多核苷酸)融合的点。
在本发明中，根据常规已知方法，基于本发明的多核苷酸的序列信息，将引物或探针制备成与本发明的多核苷酸特异性杂交的多核苷酸。引物或探针的碱基数为10-50，优选15-50，更优选18-35。
引物或探针不需要完全互补，只要该引物或探针与本发明的多核苷酸特异性杂交即可。引物或探针是与相应碱基序列具有至少70％同一性、优选至少80％同一性、更优选至少90％同一性、更优选至少95％同一性、更优选至少98％同一性的多核苷酸。
本发明的引物或探针优选为(i)SEQ ID NO：69、(ii)SEQ ID NO：70或(iii)SEQ IDNO：71所示的多核苷酸，更优选为作为一组(iv)SEQ ID NO：69和SEQ ID NO：70所示的有义引物和反义引物的多核苷酸，更优选为作为一组(v)SEQ ID NO：69、SEQ ID NO：70和SEQ IDNO：71所示的有义引物、反义引物和探针的多核苷酸。
本发明包括检测样品中本发明的多核苷酸的存在的方法。
在本发明中，检测样品中本发明的多核苷酸的存在的方法是根据常规使用检测方法例如Northern印迹、Southern印迹、RT-PCR、实时PCR、数字PCR、DNA芯片、原位杂交和序列分析的检测方法。
在本发明中，检测样品中本发明的多核苷酸的存在的方法还包括检测包括本发明的多核苷酸的RET融合基因的多核苷酸的存在的方法。该方法包括对通过使用与编码RET蛋白质的多核苷酸杂交的引物(例如，与来自RET激酶结构域的3'侧的序列杂交的引物)的5'RACE技术扩增的PCR产物进行序列分析的方法。
在本发明中，检测样品中本发明的多核苷酸的存在的方法优选地包括使用本发明的引物或探针检测本发明的多核苷酸的步骤。
本发明包括用于治疗对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症的药物组合物，该组合物包括抑制RET的化合物作为活性成分。
更优选地，本发明包括用于治疗对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症的药物组合物，该组合物包括抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物作为活性成分。
在本发明中，术语“对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症”中的“对于DCTN1基因和RET基因的融合基因呈阳性的癌症”是指表达本发明的多核苷酸的癌症，优选地是指其中已经使用检测本发明的多核苷酸的存在的方法检测到本发明的多核苷酸的癌症。
在本发明中，术语“对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症”中的“对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症”是指表达本发明的多肽的癌症，优选地是指其中已经使用检测本发明的多肽的存在的方法检测到本发明的多肽的癌症。
根据本发明的用于癌症治疗的药物组合物的活性成分是抑制RET的化合物，更优选为抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物。也可以使用通过本发明的筛选方法选择的化合物作为活性成分。例如，可以使用已知抑制RET的化合物作为本发明的药物组合物的活性成分。抑制RET的化合物可以是抑制其他酪氨酸激酶的表达和/或活性的化合物，只要化合物可以抑制RET的表达和/或活性，更优选为抑制RET的活性和其他酪氨酸激酶的表达和/或活性的化合物。此类化合物的实例包括凡德他尼(vandetanib)、索拉非尼(sorafenib)、舒尼替尼(sunitinib)、莫替塞尼(motesanib)、卡博替尼(cabozantinib)、乐伐替尼(lenvatinib)，以及在WO2016/127074小册子、WO2017/043550小册子、WO2017/011776小册子和WO2017/146116小册子中描述的化合物。
用于治疗对于DCTN1基因和RET基因的融合基因呈阳性和/或对于DCTN1蛋白和RET蛋白的融合蛋白呈阳性的癌症的药物组合物的活性成分是抑制RET的化合物；更优选为凡德他尼、卡博替尼、乐伐替尼、WO2017/043550小册子中公开的式(1)表示的稠合嘧啶化合物和WO2017/146116小册子中公开的式(1)表示的稠合嘧啶化合物；更优选为凡德他尼、卡博替尼、乐伐替尼、在WO2017/043550小册子中公开的实施例化合物1-90和WO2017/146116小册子中公开的实施例化合物1-207；再更优选为凡德他尼、卡博替尼、乐伐替尼、4-氨基-1-(叔丁基)-N-(5-甲基-1H-吡唑-3-基)-1H-吡唑并[3,4-d]嘧啶-3-甲酰胺、4-氨基-7-(叔丁基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-7-(1-氟-2-甲基丙-2-基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-N-(5-甲基-1H-吡唑-3-基)-7-(1-甲基环丙基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-7-(2-环丙基丙-2-基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-(3-吗啉代丙-1-炔-1-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-((四氢-2H-吡喃-4-基)乙炔基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、(R)-4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-((四氢呋喃-2-基)甲氧基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺和4-氨基-N-[4-(甲氧基甲基)苯基]-6-((1-甲基-1H-吡唑-4-基)乙炔基)-7-(1-甲基环丙基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；并且特别优选为4-氨基-1-(叔丁基)-N-(5-甲基-1H-吡唑-3-基)-1H-吡唑并[3,4-d]嘧啶-3-甲酰胺、4-氨基-7-(叔丁基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-7-(1-氟-2-甲基丙-2-基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-N-(5-甲基-1H-吡唑-3-基)-7-(1-甲基环丙基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-7-(2-环丙基丙-2-基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-(3-吗啉代丙-1-炔-1-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、4-氨基-N-[4-((甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-((四氢-2H-吡喃-4-基)乙炔基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺、(R)-4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-((四氢呋喃-2-基)甲氧基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺和4-氨基-N-[4-(甲氧基甲基)苯基]-6-((1-甲基-1H-吡唑-4-基)乙炔基)-7-(1-甲基环丙基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺。
在本发明中，例如，术语“化合物可以抑制RET的表达和/或活性”中的“可以抑制RET的表达”是指以下含义。使表达RET的多肽和/或多核苷酸的细胞与测试化合物接触，并检测细胞中RET的多肽或多核苷酸的表达水平。当接触过测试化合物的细胞与没有接触过测试化合物的细胞相比表现出更低的RET的多肽或多核苷酸表达水平时，确定RET的表达受到抑制。此类化合物包括上述化合物、siRNA、miRNA和核酸(DNA、RNA)适体。siRNA的实例包括CACAUGUCAUCAAAUUGUATT(SEQ ID NO：74)、GGAUUGAAAACAAACUCUATT(SEQ ID NO：75)和GCUUGUCCCGAGAUGUUUATT(SEQ ID NO：76)；并且siRNA优选为CACAUGUCAUCAAAUUGUATT(SEQID NO：74)或GGAUUGAAAACAAACUCUATT(SEQ ID NO：75)。
术语“化合物可以抑制RET的表达和/或活性”中化合物是否“可以抑制RET的活性”可以使用酪氨酸磷酸化作为指标来确定。测量酪氨酸磷酸化的方法的实例包括WO2017/043550小册子中的测试例1中所述的方法。
另外，使用表达RET的多肽和/或多核苷酸的细胞，以细胞生长抑制作用作为指标，可以确定化合物能够抑制RET的活性。测量细胞生长抑制作用的方法的实例包括WO2017/043550小册子中的测试例3和测试例4中所述的方法。
本发明的药物组合物靶向的癌症没有特别限定，只要癌症表达本发明的多核苷酸和/或多肽即可，实例包括头颈癌、甲状腺癌、胃肠道癌(例如，食道癌、胃癌、十二指肠癌、肝癌、胆道癌(例如胆囊癌和胆管癌)、胰腺癌、小肠癌、肠癌(例如结直肠癌、结肠癌、直肠癌)和胃肠道间质瘤)、肺癌(非小细胞肺癌、小细胞肺癌)、乳腺癌、卵巢癌、子宫癌(例如宫颈癌、子宫内膜癌)、肾癌、膀胱癌、前列腺癌和皮肤癌。癌症优选为甲状腺癌或肺癌(非小细胞肺癌、小细胞肺癌)。如本文所用，癌症不仅包括原发性肿瘤，还包括已经扩散到其他器官(例如肝)的癌症。
含有抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物作为活性成分的制剂可以制备成含有药物载体的药物组合物的形式，以适合多种剂型。剂型的实例包括口服剂、注射剂、栓剂、油膏和贴剂。这些剂型可以通过本领域技术人员已知和通用的制备方法来制备。
使用的药物载体包括通常用作制剂材料的各种有机或无机载体物质，并且这些载体作为固体制剂的赋形剂、粘合剂、崩解剂、润滑剂、包衣剂等添加；作为液体制剂的溶剂、增溶剂、悬浮剂、张度剂(tonicity agent)、pH调节剂和缓冲剂、舒缓剂等添加。任选地可以使用的是用于制剂的添加剂，例如防腐剂、抗氧化剂、着色剂、调味剂和稳定剂。
在制备口服固体制剂时，将赋形剂(可选地与赋形剂、粘合剂、崩解剂、润滑剂、着色剂、调味剂等)添加到本发明的化合物中，然后根据常规方法制造片剂、包衣片剂、颗粒剂、散剂、胶囊剂等。
在制备口服液体制剂时，将pH调节剂和缓冲剂、稳定剂、调味剂等添加到本发明的化合物中，并且根据常规方法制造内部液体药物、糖浆药物、酏剂等。
在制备注射剂时，将pH调节剂和缓冲剂、稳定剂、张度剂、局部麻醉剂等添加到本发明的化合物中，然后根据常规方法制造皮下注射剂、肌肉内注射剂或静脉内注射剂。
本发明包括通过检测本发明的多肽的存在的方法或者通过检测本发明的多核苷酸的存在的方法检测到样品中存在本发明的多肽或本发明的多核苷酸的存在时诊断癌症的方法。在本发明中诊断的癌症包括列为本发明的药物组合物的靶标的那些癌症。如上所述，使用本发明的多肽或多核苷酸能够诊断癌症。因此，本发明的多肽和多核苷酸可以用作检测癌症的生物标志物。
本发明包括使用本发明的多肽或本发明的多核苷酸作为确定使用抑制RET的化合物的化学疗法是否有效的指标的方法，当通过本发明的检测方法在样品中检测到本发明的多肽时，和/或当通过本发明的检测方法在样品中检测到本发明的多核苷酸的存在时，该方法确定使用抑制RET的化合物的化学疗法是有效的。
更优选地，本发明包括使用本发明的多肽或本发明的多核苷酸作为确定使用抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物的化学疗法是否有效的指标的方法，当通过本发明的检测方法在样品中检测本发明的多肽时，和/或当通过本发明的检测方法在样品中检测到本发明的多核苷酸的存在时，该方法确定使用抑制本发明的多肽的表达和/或活性或者本发明的多核苷酸的表达的化合物的化学疗法是有效的。
更优选地，本发明包括使用本发明的多肽或本发明的多核苷酸作为确定使用本发明的筛选方法中获得的化合物的化学疗法是否有效的指标的方法，当通过本发明的检测方法在样品中检测到本发明的多肽时，和/或当通过本发明的检测方法在样品中检测到本发明的多核苷酸的存在时，该方法确定使用本发明的筛选方法中获得的化合物的化学疗法是有效的。
以下实施例详细描述本发明。但是，本发明不限于这些实施例。
实施例
实施例1：DCTN1基因与RET基因的融合基因(DCTN1-RET融合基因)的制备
1-1：源自临床样本的RNA的提取
根据以下方法，使用RNeasy Mini试剂盒(Qiagen)从购自Asterand Bioscience的人甲状腺癌组织中提取RNA。将600μL的缓冲液RLT加入至甲状腺癌组织，并施加至QIAshredder旋转柱，随后离心(16,000rpm，2分钟，室温)，从而收集滤液。将等量的70％乙醇水溶液加入到收集的滤液中。将其混合之后，将混合物加到RNeasy Mini柱上，然后离心(10,000rpm，15秒，室温)。将700μL Buffer RW1加入至RNeasy Mini柱中，并进行离心(10,000rpm，15秒，室温)。向其中进一步加入500μL Buffer RPE，并离心(10,000rpm，15秒，室温)。以相同的方式，再次加入500μL Buffer RPE，并离心(10,000rpm，2分钟，室温)。将RNeasy Mini柱再次离心(16,000rpm，1分钟，室温)，然后除去剩余的缓冲液。将40μL不含RNase的水加到RNeasy Mini柱上并离心(10,000rpm，1分钟，室温)，从而收集滤液作为总RNA。
1-2：源自临床样本的cDNA的制备
根据以下方法，使用SuperScript VILO cDNA合成试剂盒(Invitrogen)从以上1-1节中获得的总RNA合成cDNA。将500ng总RNA用不含RNase的水调节，得到14μL的量，并向其中加入4μL 5×VILO Reaction Mix和2μL 10×SuperScript酶混合物，并混合。将混合物在25℃下保温10分钟，然后在42℃下保温60分钟。为了终止反应，最后将混合物在85℃下孵育5分钟，从而获得cDNA。
1-3：克隆载体的制备和提纯
为了扩增DCTN1-RET融合基因，设计表1所示的引物：引物1(SEQ ID NO：33)作为有义引物，引物2(SEQ ID NO：34)作为反义引物，以及引物3(SEQ ID NO：35)作为有义引物，而引物4(SEQ ID NO：36)作为反义引物，用于巢式(nested)PCR。
表1
引物1 5'-TGTCCAGCTTTGTGCCTGATTGATGT-3' SEQ ID NO:33
引物2 5'-GCTGGGCACTGAAGAGAAAGGAATGC-3' SEQ ID NO:34
引物3 5'-AGCAGGATGAGTGCGGAGGCAAGC-3' SEQ ID NO:35
引物4 5'-TTAACTATCAAACGTGTCCATTAATTTTGCCGC-3' SEQ ID NO:36
根据以下方法，使用这些引物并使用KOD-Plus-Neo(Toyobo)，以上文1-2节中合成的cDNA为模板，扩增DCTN1-RET融合基因。将2μL cDNA、5μL用于KOD-Plus-Neo的10×PCR缓冲液、5μL 2mM dNTP、3μL 25mM MgSO4、1μL KOD-Plus-Neo、1.5μL引物1(10μM)、1.5μL引物2(10μM)和31μL双重蒸馏水(DDW)混合；进行PCR。随后，将获得的PCR产物稀释100倍，然后将2μL稀释的PCR产物、5μL用于KOD-Plus-Neo的10×PCR缓冲液、5μL 2mM dNTPs、3μL 25mMMgSO4、1μL KOD-Plus-Neo、1.5μL引物3(10μM)、1.5μL引物4(10μM)和31μL DDW混合；并进行巢式PCR。
使用1％琼脂糖凝胶(Nacalai Tesque)通过电泳分离巢式PCR产物，并使用QIAquick Gel Extraction试剂盒(Qiagen)从凝胶中提纯PCR产物。
将通过限制酶SmaI(NEB)切割的pUC18 DNA(Takara Bio Inc.)、提纯的PCR产物、T4 DNA连接酶(NEB)和T4 DNA连接酶反应缓冲液(NEB)混合，并将混合物在16℃下孵育过夜。将连接产物用SmaI(NEB)处理，并通过以下方法进行感受态细胞的转化。将用SmaI处理过的连接产物加入到50μL的大肠杆菌DH5α感受态细胞(Takara Bio Inc.)中，将其在冰上静置30分钟。之后，将细胞在42℃下热激30秒，并使其在冰上静置2分钟。向其中加入SOC培养基(Takara Bio Inc.)，并将细胞在37℃下振荡培养1小时。然后将培养液施加到含有氨苄青霉素的LB琼脂培养基平板(Unitech)上，并在37℃下放置过夜。将大肠杆菌菌落悬浮在含有氨苄青霉素的LB培养基(InvivoGen)中，并在37℃下振荡培养过夜。使用QIAquickSpin Miniprep试剂盒(Qiagen)，根据试剂盒提供的方案，从增殖的大肠杆菌中提纯插入DCTN1-RET融合基因的质粒DNA。
1-4：序列的测定
使用上文1-3节中获得的质粒DNA作为模板，使用表2所示的用于测序的引物5-36，并使用BigDye Terminator V3.1循环测序试剂盒进行序列反应；使用Applied Biosystems3730xl DNA分析仪进行序列分析。序列分析的结果表明，DCTN1-RET融合基因是其中RET变体2(GenBank登录号：NM_020975)的外显子12-20融合在DCTN1变体5(GenBank登录号：NM_001190836)的外显子1-27的3'侧下游的基因(SEQ ID NO：17)。
表2
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实施例2：DCTN1-RET融合基因的检测
使用SuperScript VILO cDNA合成试剂盒(Invitrogen)，根据以下方法，从购自Asterand Bioscience公司的源自正常人甲状腺组织的RNA和1-1节中获得的源自人甲状腺癌组织的RNA合成cDNA。将280ng总RNA用不含RNase的水调节，以得到14μL的量，并且分别向其中加入4μL 5×VILO Reaction Mix和2μL 10×SuperScript酶混合物，并混合。将混合物在25℃下保温10分钟，然后在42℃下保温60分钟。为了终止反应，最后将混合物在85℃下孵育5分钟。
为了检测DCTN1-RET融合基因，设计表3所示的引物和探针：引物37(SEQ ID NO：69)作为用于检测DCTN1-RET融合基因的有义引物，引物38(SEQ ID NO：70)作为用于检测DCTN1-RET融合基因的反义引物，以及引物39作为用于检测DCTN1-RET融合基因的探针(SEQID NO：71)(探针：TaqMan MGB探针；荧光染料：FAM(Thermo Fisher Scientific))。
表3
引物37 5'-CTGGAGCCACAGTACCCACT-3' SEQ ID NO:69
引物38 5'-TCCAAATTCGCCTTCTCCTA-3' SEQ ID NO:70
引物39 5'-TTCATCAGCCTTCCTCAGGGAGGAT-3' SEQ ID NO:71
将获得的cDNA稀释10倍，并将其1.1μL用作模板。将11μL用于探针的ddPCRSupermix(Bio-Rad)、2μL引物37(10μM)、2μL引物38(10μM)、0.6μL引物39(10μM)和1.1μL用于检测GAPDH的20×HEX assay(Prime PCR ddPCR Expression Probe Assay：GAPDH，Human，Bio-Rad)；使用自动液滴发生器(Bio-Rad)制备液滴。对制备的液滴进行PCR，并用液滴读数器(Bio-Rad)对DCTN1-RET和GAPDH阳性的液滴进行计数。图1和2显示了结果。
根据以下方法，使用KOD-Plus-Neo(Toyobo)以上述合成的cDNA为模板，扩增DCTN1-RET融合基因。将2μL cDNA、5μL用于KOD-Plus-Neo的10×PCR缓冲液、5μL 2mM dNTP、3μL 25mM MgSO4、1μL KOD-Plus-Neo、1.5μL引物1(10μM)、1.5μL引物2(10μM)和31μL DDW混合，并进行PCR。随后，将获得的PCR产物稀释100倍，然后将2μL稀释的PCR产物、5μL用于KOD-Plus-Neo的10×PCR缓冲液、5μL 2mM dNTPs、3μL 25mM MgSO4、1μL KOD-Plus-Neo、1.5μL引物3(10μM)、1.5μL引物4(10μM)和31μL DDW混合；并进行巢式PCR。使用1％琼脂糖凝胶(Nacalai Tesque)通过电泳分离巢式PCR产物，并照相。结果如图3所示。
如图1、图2和图3所示，在由源自人甲状腺癌组织的RNA合成的cDNA中检测到DCTN1-RET融合基因，而在由源自正常人的甲状腺组织的RNA合成的cDNA中没有检测到DCTN1-RET融合基因。这些结果表明DCTN1-RET融合基因可用作癌症的生物标志物。
实施例3：DCTN1-RET融合基因表达载体的构建
为了构建表达载体，如表4所示设计引物40(SEQ ID NO：72)作为有义引物，引物41(SEQ ID NO：73)作为反义引物。
表4
根据下述方法，使用这些引物、上文1-2节中合成的cDNA作为模板以及Prime STARMax DNA聚合酶(TaKaRa)扩增DCTN1-RET融合基因。将1μL cDNA、25μL 2×Prime STAR MaxDNA聚合酶、1μL引物40(10μM)、1μL引物41(10μM)和22μL双重蒸馏水(DDW)混合，以进行PCR。使用1％琼脂糖凝胶(Nacalai Tesque)通过电泳分离得到的PCR产物，并使用GFX PCR DNA和凝胶带提纯试剂盒(GE Healthcare)从凝胶提纯PCR产物。随后，根据下述方法，使用Gateway BP Clonase II酶混合物(Thermo Fisher)将提纯的PCR产物插入GatewaypDONR221载体中，从而制备入门载体(entry vector)。具体而言，将5.0μL提纯的PCR产物、3.5μL pDONR221(85ng/μL)、4.0μL BP Clonase II酶混合物和7.5μL TE混合，并在25℃下孵育90分钟。温育之后，向其中加入1μL蛋白酶K(2mg/mL)，然后在37℃下孵育10分钟，从而制备入门载体。
将获得的入门载体加入到50μL大肠杆菌DH5α感受态细胞(Takara Bio Inc.)中，并在冰上静置30分钟。之后，将细胞在37℃下热激20秒，并使其在冰上静置2分钟。然后向其中加入SOC培养基(Takara Bio Inc.)，并将细胞在37℃下振荡培养1小时。然后将培养液施加到含有卡那霉素的LB琼脂培养基板上，并在37℃下静置过夜。将大肠杆菌菌落悬浮在含有卡那霉素的LB培养基中，并在37℃下振荡培养过夜。用GENE PREP STAR PI-480自动DNA分离系统(Kurabo Industries Ltd.)从生长的大肠杆菌中提纯DCTN1-RET融合基因插入质粒DNA(入门载体克隆)。
使用获得的质粒和Gateway LR Clonase II酶混合物(Thermo Fisher)，根据下述方法将DCTN1-RET融合基因插入pJTI Fast DEST载体，从而制备表达载体。将150ng入门载体克隆、1μL pJTI Fast DEST载体(150ng/μL)、2μL LR Clonase II酶混合物和TE缓冲液混合，总共得到10μL，并将混合物在25℃下孵育90分钟。孵育之后，向其中加入1μL蛋白酶K(2mg/mL)，然后在37℃下孵育10分钟，从而获得插入DCTN1-RET融合基因的pJTI Fast DEST载体(DCTN1-RET融合基因表达载体)。将获得的DCTN1-RET融合基因表达载体加入到50μL大肠杆菌DH5α感受态细胞(Takara Bio Inc.)中，并在冰上静置30分钟。此后，将细胞在37℃下热激20秒，并使其在冰上静置2分钟。然后向其中加入SOC培养基(Takara Bio Inc.)，并将细胞在37℃下振荡培养1小时。将培养液施加在含有氨苄青霉素的LB琼脂培养基板上，在37℃下放置过夜。将大肠杆菌菌落悬浮在含有氨苄青霉素的LB培养基中，并在37℃下振荡培养过夜。使用Plasmid Plus Maxi试剂盒(QIAGEN)由生长的大肠杆菌提纯插入DCTN1-RET融合基因的质粒DNA(DCTN1-RET融合基因表达载体)。
实施例4：DCTN1-RET融合基因表达细胞的建立
4-1：细胞的建立
对于建立DCTN1-RET融合基因表达细胞的宿主细胞，选择小鼠胚胎成纤维细胞NIH/3T3细胞(美国典型培养物保藏中心(American Type Culture Collection))，并用上文制备的插入DCTN1-RET融合基因的表达载体转染细胞，从而建立表达DCTN1-RET融合基因的细胞。具体步骤如下。NIH/3T3细胞通过在5％CO2中在37℃下在用于典型培养(二维细胞培养)的培养基中培养而制备；通过将新生小牛血清(NBCS)(Gibco)加入到D-MEM(高葡萄糖)(含有L-谷氨酰胺、酚红、丙酮酸钠、1500mg/L碳酸氢钠)(Wako)中至10％，制备用于二维细胞培养的培养基。在进行转染的前一天，将NIH/3T3细胞以1.5×105细胞/2mL接种到6孔板(Iwaki)中，并在5％CO2中在37℃下孵育过夜。将ViaFect转染试剂加入到通过将1.5μgDCTN1-RET融合基因表达载体和1.5μg pJTI phiC31整合酶载体混合制备的混合溶液中，使得ViaFect转染试剂的量是混合溶液量的六倍；然后向其中加入Opti-MEM，使其总量为300μL，然后在室温下孵育5分钟，从而制备转染溶液。从接种有NIH/3T3细胞的孔中，除去300μL培养基，并将300μL制备的转染溶液加入到孔中，然后在5％CO2中在37℃下孵育过夜。次日，更换培养基以去除转染溶液。更换培养基时，将潮霉素B(Nacalai Tesque)加入到新的培养基中，达到500μg/mL。潮霉素B去除了未转染DCTN1-RET融合基因插入表达载体的细胞。转染之后，在每周约两次更换培养基的同时，培养细胞直至其增殖。转染22天后，用胰蛋白酶收集细胞，并根据以下方法进行单细胞克隆。测量收集的细胞数，并向其中添加培养基，达到1细胞/200μL。将细胞接种到96孔板(Thermo Fisher)上，达到每孔200μL。接种之后，每天观察细胞，并取出从单个细胞生长的细胞。这些细胞是表达DCTN1-RET融合基因的细胞(表达DCTN1-RET融合基因的NIH/3T3细胞)。
4-2：确认靶向蛋白质的表达
通过蛋白质免疫印迹确认DCTN1-RET融合蛋白在获得的表达DCTN1-RET融合基因的NIH/3T3细胞中的表达。具体地，从培养瓶中除去培养基，然后用PBS洗涤一次。将含有磷酸酶抑制剂(Roche)和蛋白酶抑制剂(Roche)的样品稀释液浓缩液2(R&D Systems)加入到培养瓶中，并用刮刀收集细胞裂解液。从收集的细胞裂解液中，通过离心获得蛋白质样品。对蛋白质样品进行蛋白质测定，并将蛋白质浓度调节至规定浓度。将用于SDS-PAGE的具有还原剂(6x)的样品缓冲溶液(Nacalai Tesque)加入到规定浓度的蛋白质样品中；然后将混合物在95℃下孵育5分钟，以使蛋白质变性，从而获得用于蛋白质免疫印迹的样品。对于阴性对照，根据相同的方法使用NIH/3T3细胞(亲代细胞系)获得用于蛋白质免疫印迹的样品。根据下述方法，使用样品确认蛋白质的表达。使用4-15％丙烯酰胺凝胶(Bio-Rad)和1×Tris/甘氨酸/SDS缓冲液通过SDS-PAGE(200V下30分钟)分离蛋白质。使用Trans-BlotTurbo RTA Midi PVDF转移试剂盒(Bio-Rad)和Trans-Blot Turbo转录系统(Bio-Rad)将蛋白质转移到PVDF膜上，然后将PVDF膜浸入Blocking One-P 1小时。通过将Blocking One-P用TBS-T稀释达到10％，制备溶液，将一抗(Phospho-RET(Tyr905)抗体(CST)、Ret(C31B4)兔mAb(CST)和抗-Dctn1抗体(Atlas Antibodies))用制备的溶液稀释，达到1/1000的浓度。将PVDF膜浸入所得溶液中，并在4℃下孵育过夜。用TBS-T洗涤之后，将PVDF膜浸入通过将抗兔IgG、HRP-连接抗体(CST)用TBS-T稀释以达到浓度1/2000而制备的二次抗体稀释溶液中；并在室温下孵育1小时。将膜用TBS-T洗涤后，使用SuperSignal West Dura ExtendedDuration底物(Thermo Fisher)和Amersham Imager 600lumino图像分析仪(GEHealthcare)对蛋白质进行检测。用Precision Plus Protein Kaleidoscope预染色蛋白质标准品(Bio-Rad)确认检测到的蛋白质的分子量。如图4a)和图4b)中所示，当使用抗-pRET抗体或抗-RET抗体时，没有检测到内源性RET(150和175kDa)。然而，仅在表达DCTN1-RET融合基因的NIH/3T3细胞中，在约175kDa处证实了似乎是DCTN1-RET融合蛋白的条带。另外，如图4c)所示，在使用抗DCTN1抗体的情况下，在约150kDa处检测到内源性DCTN1。仅在表达DCTN1-RET融合基因的NIH/3T3细胞中，在约150kDa处的内源性DCTN1的条带的上方还检测到约175kDa的条带。具体地，仅在表达DCTN1-RET融合基因的NIH/3T3细胞中，在使用抗RET抗体和抗DCTN1抗体时均检测到约175kDa的条带。这清楚地表明，在制备的表达DCTN1-RET融合基因的NIH/3T3细胞中表达了DCTN1和RET的融合蛋白。
实施例5：通过三维细胞培养证实表达DCTN1-RET融合基因的NIH/3T3细胞的生长
NIH/3T3细胞在二维细胞培养条件下生长良好，但在三维细胞培养条件下几乎不生长。然而，当在细胞中表达癌基因时，已知NIH/3T3细胞也可以在三维细胞培养条件下生长。因此，使用该特征确认了DCTN1-RET融合基因是否是癌基因。将表达DCTN1-RET融合基因的NIH/3T3细胞和NIH/3T3细胞在5％CO2中在37℃下在二维细胞培养物中培养；并用胰蛋白酶收集，然后对细胞数量进行计数。为了进行三维细胞培养，使用FCeM系列制备试剂盒(Nissan Chemical Industries,Ltd.)、D-MEM(高葡萄糖)(含有L-谷氨酰胺、酚红、丙酮酸钠和1500mg/L碳酸氢钠)(Wako)和新生小牛血清(NBCS)(Gibco)制备用于三维细胞培养的培养基。将细胞悬浮在制备的用于三维细胞培养的培养基中，以得到1000个细胞/90μL，并以每孔90μL接种到96孔透明黑色圆底球形微孔板(Corning)中，然后在5％CO2中在37℃下孵育。接种后下一天(第1天)和接种后8天(第8天)后，使用用于细胞内ATP发光的检测试剂(CellTiter-Glo 2.0试剂，Promega)用发光计(EnSpire，PerkinElmer)测量发光水平(每秒计数：cps)，并将结果确定为活细胞计数的指标。然后从第1天的测量结果和第8天的测量结果计算细胞生长速率(N＝3)。结果如图5所示，第8天的NIH/3T3细胞的细胞数量是第1天的细胞数量的2.4倍，而第8天的表达DCTN1-RET融合基因的NIH/3T3细胞的细胞数量是第1天的细胞数量的20.9倍。此外，尽管通过三维细胞培养在NIH/3T3细胞中没有形成细胞聚集体，但确认通过三维细胞培养在表达DCTN1-RET融合基因的NIH/3T3细胞中形成细胞聚集体。这清楚地表明，DCTN1-RET融合基因的转染增强了细胞的生长，表明DCTN1-RET融合基因是癌基因。
实施例6：证实表达DCTN1-RET融合基因的NIH/3T3细胞在体内的致瘤性
为了证实表达DCTN1-RET融合基因的NIH/3T3细胞在体内的致瘤性，对裸鼠进行了移植实验。NIH/3T3细胞是亲本细胞系，已知在裸鼠中不会皮下生长；通过将裸鼠皮下移植表达DCTN1-RET融合基因的NIH/3T3细胞，可以证实DCTN1-RET融合基因是否有助于致癌性，或者DCTN1-RET融合基因是否是癌基因。使用裸鼠BALB/cAJcl-nu/nu(CLEA Japan，Inc.)进行移植。将表达DCTN1-RET融合基因的NIH/3T3细胞用胰蛋白酶收集，并悬浮在PBS中，最终得到1×108细胞/mL。向其中加入等量的Matrigel Basement Membrane Matrix(Corning)，并调节至5×107细胞/mL，制备用于移植的细胞悬液。使用25G注射针和1-mL注射器将0.1mL用于移植的细胞悬液皮下移植到每只裸鼠(N＝10)的右侧胸部。在移植后的第10、13和17天，用数字卡尺(Mitutoyo Corporation)测量每只小鼠的肿瘤的长轴和短轴，并根据以下方程式计算肿瘤体积。
肿瘤体积(mm3)＝(长轴：mm)×(短轴：mm)×(短轴：mm)/2
图6示出肿瘤体积的测量结果。在裸鼠皮下移植的表达DCTN1-RET融合基因的NIH/3T3细胞被证实具有肿瘤发生能力并且生长良好。因此，该体内实验还表明DCTN1-RET融合基因是癌基因。
实施例7：在表达DCTN1-RET融合基因的NIH/3T3细胞中证实siRNA对DCTN1-RET融合蛋白的抑制和细胞生长的抑制作用
考察了siRNA处理对表达DCTN1-RET融合基因的NIH/3T3细胞的影响。使用的siRNA如下：下表5所示的三种类型的RET siRNA，以及Silencer Select阴性对照#1siRNA(Ambion)作为阴性对照。三种类型的RET siRNA的靶标是人RET；RET siRNA1和RET siRNA2含有与DCTN1-RET融合基因的RET部分结合的序列，而RET siRNA3不含与DCTN1-RET融合基因的结合序列。换句话说，RET siRNA1和RET siRNA2据信抑制DCTN1-RET融合基因的表达，而RET siRNA3则不会。下面描述使用siRNA的实验方法。
表5
在5％CO2中在37℃下通过在用于二维细胞培养的培养基中培养，制备表达DCTN1-RET融合基因的NIH/3T3细胞。在进行siRNA处理的前一天，将细胞接种到6孔板(Iwaki)中，3×105细胞/2mL，并在5％CO2中在37℃下孵育过夜。将12μL预先用水调节至20μM的每种siRNA、4μL的Lipofectamine RNAiMAX转染试剂(Thermo Fisher)和384μL的Opti-MEM混合；并在室温下孵育15分钟，从而制备siRNA溶液。将400μL每种siRNA溶液加入到其中接种有细胞的孔中，并在5％CO2中在37℃下孵育过夜。次日，将一部分培养的细胞作为样品用于蛋白质表达分析；另取一部分重新接种以确认细胞生长。以与上文所述的4-2：确认靶向蛋白质的表达中相同的方式进行蛋白质表达分析取样和蛋白质表达分析，不同之处在于使用Phospho-RET(Tyr905)抗体(CST)、Ret(C31B4)兔mAb(CST)和GAPDH(D16H11)XP兔mAb(CST)作为一抗。结果表明，如图7所示，与未经siRNA处理(未处理)的细胞相比，在用阴性对照siRNA(NC)处理的细胞中DCTN1-RET融合蛋白的表达没有受到抑制。当用RET siRNA1或RETsiRNA2处理细胞时，证实DCTN1-RET融合蛋白的表达在表达DCTN1-RET融合基因的NIH/3T3细胞中已被抑制，而在使用RET siRNA3时未被抑制。
随后，为了确认细胞生长抑制作用，从未经处理的孔和经siRNA处理的孔中用胰蛋白酶收集细胞，并计数细胞数。然后以与实施例5相同的方式进行三维细胞培养，并且在接种的当天(第0天)和从接种的第4天后(第4天)，以与实施例5相同的方式对活细胞数进行计数。从第0天的测量结果和第4天的测量结果计算细胞的生长速率。结果表明，如图8所示，未经siRNA处理(未经处理)的细胞和经阴性对照siRNA(NC)处理的细胞在第4天的细胞数分别是第0天的4.9倍和3.6倍；然而，用RET siRNA1处理的细胞和用RET siRNA2处理的细胞分别仅表现出约2.0倍和2.4倍的生长，两者均明显较低。用RET siRNA3处理的细胞显示出3.7倍的生长，基本等于用阴性对照siRNA处理的细胞的生长，很少显示出生长降低。这些结果表明当siRNA抑制RET的表达时，表达DCTN1-RET融合基因的NIH/3T3细胞的生长也被抑制。
实施例8：表达DCTN1-RET融合基因的NIH/3T3细胞中的细胞生长抑制作用
在表达DCTN1-RET融合基因的NIH/3T3细胞上进行了体外细胞生长检验。以与实施例5相同的方式进行三维细胞培养和接种。接种后，在5％CO2中在37℃下孵育过夜(第0天)。将据报道能够抑制RET的卡博替尼、凡德他尼、艾乐替尼(alectinib)、乐伐替尼和稠合嘧啶化合物(表6所示的化合物1-9)溶于二甲亚砜中，浓度为10mmol/L；并进一步用三维细胞培养用培养基稀释，使得这些化合物的终浓度为1000nmol/L、333nmol/L、111nmol/L、37.0nmol/L、12.3nmol/L、4.12nmol/L、1.37nmol/L和0.457nmol/L。然后将这些稀释的化合物分别加入到其中接种有细胞的板的每个孔中(每孔0.01mL)(第1天)；将细胞在5％CO2中在37℃下孵育7天。培养之后(第8天)，将细胞内ATP发光检测试剂(CellTiter-Glo 2.0试剂，Promega)加入到每个孔中，并用发光计(EnSpire，PerkinElmer)测量发光水平(每秒计数：cps)。基于以下方程，根据Tday8和Cday1的值，以及每种测试化合物的细胞生长被抑制50％的浓度(GI50(nM))，计算从第1天起用不同浓度的化合物处理的细胞的生长速率。
1)当Tday8≥Cday1
生长速率(％)＝(Tday8-Cday1)/(Cday8-Cday1)×100
T：加入测试化合物的孔的cps
C：未加入测试化合物的孔的cps
day1：加入测试化合物的当天
day8：进行评估的当天
2)当Tday 8<Cday1
生长速率(％)＝(Tday8-Cday1)/(Cday1)×100
T：加入测试化合物的孔的cps
C：未加入测试化合物的孔的cps
day1：加入测试化合物的当天
day8：进行评估的当天
表6
续表6
结果表明，如表7所示，卡博替尼、凡德他尼、乐伐替尼和稠合嘧啶化合物(化合物1-9)抑制了表达DCTN1-RET融合基因的NIH/3T3细胞的生长。
表7
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以上结果表明了这些RET抑制剂作为其中已检测到DCTN1-RET融合基因的癌症的治疗剂的潜在用途。结果还表明，使用表达DCTN1-RET融合基因的NIH/3T3细胞能够筛选抑制DCTN1-RET的化合物。
实施例9：表达DCTN1-RET融合基因的细胞中RET磷酸化的抑制
以下根据下述方法考察表达DCTN1-RET融合基因的细胞中的RET磷酸化是否被经报道抑制RET的现有药物所抑制。通过在5％CO2中在37℃下在用于二维细胞培养的培养基中进行培养，制备使用的表达DCTN1-RET融合基因的NIH/3T3细胞。在用药物处理细胞的前一天，将细胞接种到6孔板(Iwaki)中，3×105细胞/2mL，并在5％CO2中在37℃下孵育过夜。将卡博替尼、凡德他尼、艾乐替尼和乐伐替尼溶解在二甲亚砜中，使其浓度为10mmol/L，并进一步用PBS稀释，使得这些化合物的终浓度为1000nmol/L、100nmol/L和10nmol/L。将稀释的化合物分别加入到预先已接种细胞的每个孔中，每孔20μL(第1天)，并在5％CO2中在37℃下孵育1小时。孵育之后，以与实施例7相同的方式获取用于蛋白质表达分析的细胞样品，并分析蛋白质表达。结果表明，如图9所示，卡博替尼和乐伐替尼显著降低了表达DCTN1-RET融合基因的NIH/3T3细胞中的磷酸化RET水平。另外，还以与上文相同的方式评价了稠合嘧啶化合物对RET磷酸化的抑制，并且还证实了RET磷酸化被稠合嘧啶化合物显著地降低。这些结果表明，能够显著降低磷酸化RET水平的药物将是可以抑制表达DCTN1-RET融合基因的NIH/3T3细胞的生长的化合物，并且该化合物潜在地可用作已检测到DCTN1-RET融合基因的癌症的治疗剂。结果还表明，使用表达DCTN1-RET融合基因的NIH/3T3细胞中的磷酸化RET的水平能够筛选RET抑制剂。
序列表自由文本
SEQ ID NO：1示出编码DCTN1变体1(v1)(SEQ ID NO：25的一部分)和RET变体2(v2)(SEQ ID NO：31的一部分)的融合肽的多核苷酸的碱基序列。
SEQ ID NO：2示出DCTN1 v1和RET v2的融合肽的氨基酸序列。
SEQ ID NO：3示出编码DCTN1 v1和RET变体4(v4)(SEQ ID NO：32的一部分)的融合肽的多核苷酸的碱基序列。
SEQ ID NO：4示出DCTN1 v1和RET v4的融合肽的氨基酸序列。
SEQ ID NO：5示出编码DCTN1变体2(v2)(SEQ ID NO：26的一部分)和RET v2的融合肽的多核苷酸的碱基序列。
SEQ ID NO：6示出DCTN1 v2和RET v2的融合肽的氨基酸序列。
SEQ ID NO：7示出编码DCTN1 v2和RET v4的融合肽的多核苷酸的碱基序列。
SEQ ID NO：8示出DCTN1 v2和RET v4的融合肽的氨基酸序列。
SEQ ID NO：9示出编码DCTN1变体3(v3)(SEQ ID NO：27的一部分)和RET v2的融合肽的多核苷酸的碱基序列。
SEQ ID NO：10示出DCTN1 v3和RET v2的融合肽的氨基酸序列。
SEQ ID NO：11示出编码DCTN1 v3和RET v4的融合肽的多核苷酸的碱基序列。
SEQ ID NO：12示出DCTN1 v3和RET v4的融合肽的氨基酸序列。
SEQ ID NO：13示出编码DCTN1变体4(v4)(SEQ ID NO：28的一部分)和RET v2的融合肽的多核苷酸的碱基序列。
SEQ ID NO：14示出DCTN1 v4和RET v2的融合肽的氨基酸序列。
SEQ ID NO：15示出编码DCTN1 v4和RET v4的融合肽的多核苷酸的碱基序列。
SEQ ID NO：16示出DCTN1 v4和RET v4的融合肽的氨基酸序列。
SEQ ID NO：17示出编码DCTN1变体5(v5)(SEQ ID 2：29的一部分)和RET v2的融合肽的多核苷酸的碱基序列。
SEQ ID NO：18示出DCTN1 v5和RET v2的融合肽的氨基酸序列。
SEQ ID NO：19示出编码DCTN1 v5和RET v4的融合肽的多核苷酸的碱基序列。
SEQ ID NO：20示出DCTN1 v5和RET v4的融合肽的氨基酸序列。
SEQ ID NO：21示出编码DCTN1 v6和RET v2的融合肽的多核苷酸的碱基序列。
SEQ ID NO：22示出DCTN1 v6和RET v2的融合肽的氨基酸序列。
SEQ ID NO：23示出编码DCTN1 v6和RET v4的融合肽的多核苷酸的碱基序列。
SEQ ID NO：24示出DCTN1 v6和RET v4的融合肽的氨基酸序列。
SEQ ID NO：33示出引物的碱基序列。
SEQ ID NO：34示出引物的碱基序列。
SEQ ID NO：35示出引物的碱基序列。
SEQ ID NO：36示出引物的碱基序列。
SEQ ID NO：37示出引物的碱基序列。
SEQ ID NO：38示出引物的碱基序列。
SEQ ID NO：39示出引物的碱基序列。
SEQ ID NO：40示出引物的碱基序列。
SEQ ID NO：41示出引物的碱基序列。
SEQ ID NO：42示出引物的碱基序列。
SEQ ID NO：43示出引物的碱基序列。
SEQ ID NO：44示出引物的碱基序列。
SEQ ID NO：45示出引物的碱基序列。
SEQ ID NO：46示出引物的碱基序列。
SEQ ID NO：47示出引物的碱基序列。
SEQ ID NO：48示出引物的碱基序列。
SEQ ID NO：49示出引物的碱基序列。
SEQ ID NO：50示出引物的碱基序列。
SEQ ID NO：51示出引物的碱基序列。
SEQ ID NO：52示出引物的碱基序列。
SEQ ID NO：53示出引物的碱基序列。
SEQ ID NO：54示出引物的碱基序列。
SEQ ID NO：55示出引物的碱基序列。
SEQ ID NO：56示出引物的碱基序列。
SEQ ID NO：57示出引物的碱基序列。
SEQ ID NO：58示出引物的碱基序列。
SEQ ID NO：59示出引物的碱基序列。
SEQ ID NO：60示出引物的碱基序列。
SEQ ID NO：61示出引物的碱基序列。
SEQ ID NO：62示出引物的碱基序列。
SEQ ID NO：63示出引物的碱基序列。
SEQ ID NO：64示出引物的碱基序列。
SEQ ID NO：65示出引物的碱基序列。
SEQ ID NO：66示出引物的碱基序列。
SEQ ID NO：67示出引物的碱基序列。
SEQ ID NO：68示出引物的碱基序列。
SEQ ID NO：69示出引物的碱基序列。
SEQ ID NO：70示出引物的碱基序列。
SEQ ID NO：71示出引物的碱基序列。
SEQ ID NO：72示出引物的碱基序列。
SEQ ID NO：73示出引物的碱基序列。
SEQ ID NO：74示出RET siRNA的碱基序列。
SEQ ID NO：75示出RET siRNA的碱基序列。
SEQ ID NO：76示出RET siRNA的碱基序列。
SEQ ID NO：77示出RET siRNA的碱基序列。
序列表
<110> 大鹏药品工业株式会社
<120> DCTN1蛋白质与RET蛋白质的融合蛋白
<130> P17-158WO
<150> JP 2017-158796
<151> 2017-08-21
<160> 77
<170> PatentIn version 3.5
<210> 1
<211> 4908
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 1
atggcacaga gcaagaggca cgtgtacagc cggacgccca gcggcagcag gatgagtgcg 60
gaggcaagcg cccggcctct gcgggtgggc tcccgtgtag aggtgattgg aaaaggccac 120
cgaggcactg tggcctatgt tggagccaca ctgtttgcca ctggcaaatg ggtaggcgtg 180
attctggatg aagcaaaggg caaaaatgat ggaactgttc aaggcaggaa gtacttcact 240
tgtgatgaag ggcatggcat ctttgtgcgc cagtcccaga tccaggtatt tgaagatgga 300
gcagatacta cttccccaga gacacctgat tcttctgctt caaaagtcct caaaagagag 360
ggaactgata caactgcaaa gactagcaaa ctgcggggac tgaagcctaa gaaggcaccg 420
acagcccgaa agaccacaac tcggcgaccc aagcccacgc gcccagccag tactggggtg 480
gctggggcca gtagctccct gggcccctct ggctcagcgt cagcaggtga gctgagcagc 540
agtgagccca gcaccccggc tcagactccg ctggcagcac ccatcatccc cacgccggtc 600
ctcacctctc ctggagcagt ccccccgctt ccttccccat ccaaggagga ggagggacta 660
agggctcagg tgcgggacct ggaggagaaa ctagagaccc tgagactgaa acgggcagaa 720
gacaaagcaa agctaaaaga gctggagaaa cacaaaatcc agctggagca ggtgcaggaa 780
tggaagagca aaatgcagga gcagcaggcc gacctgcagc ggcgcctcaa ggaggcgaga 840
aaggaagcca aggaggcgct ggaggcaaag gaacgctata tggaggagat ggctgatact 900
gctgatgcca ttgagatggc cactttggac aaggagatgg ctgaagagcg ggctgagtcc 960
ctgcagcagg aggtggaggc actgaaggag cgggtggacg agctcactac tgacttagag 1020
atcctcaagg ctgagattga agagaagggc tcagatggcg ctgcatccag ttatcagctc 1080
aagcagcttg aggagcagaa tgcccgcctg aaggatgccc tggtgaggat gcgggatctt 1140
tcttcctcag agaagcagga gcatgtgaag ctccagaagc tcatggaaaa gaagaaccaa 1200
gagctggaag ttgtgaggca acagcgggag cgtctgcagg aggagctaag ccaggcagag 1260
agcaccattg atgagctcaa ggagcaggtg gatgctgctc tgggtgctga ggagatggtg 1320
gagatgctga cagatcggaa cctgaatctg gaagagaaag tgcgcgagtt gagggagact 1380
gtgggagact tggaagcgat gaatgagatg aacgatgagc tgcaggagaa tgcacgtgag 1440
acagaactgg agctgcggga gcagctggac atggcaggcg cgcgggttcg tgaggcccag 1500
aagcgtgtgg aggcagccca ggagacggtt gcagactacc agcagaccat caagaagtac 1560
cgccagctga ccgcccatct acaggatgtg aatcgggaac tgacaaacca gcaggaagca 1620
tctgtggaga ggcaacagca gccacctcca gagacctttg acttcaaaat caagtttgct 1680
gagactaagg cccatgccaa ggcaattgag atggaattga ggcagatgga ggtggcccag 1740
gccaatcgac acatgtccct gctgacagcc ttcatgcctg acagcttcct tcggccaggt 1800
ggggaccatg actgcgttct ggtgctgttg ctcatgcctc gtctcatttg caaggcagag 1860
ctgatccgga agcaggccca ggagaagttt gaactaagtg agaactgttc agagcggcct 1920
gggctgcgag gagctgctgg ggagcaactc agctttgctg ctggactggt gtactcgctg 1980
agcctgctgc aggccacgct acaccgctat gagcatgccc tctctcagtg cagtgtggat 2040
gtgtataaga aagtgggcag cctgtaccct gagatgagtg cccatgagcg ctccttggat 2100
ttcctcattg aactgctgca caaggatcag ctggatgaga ctgtcaatgt ggagcctctc 2160
accaaggcca tcaagtacta tcagcatctg tacagcatcc accttgccga acagcctgag 2220
gactgtacta tgcagctggc tgaccacatt aagttcacgc agagtgctct ggactgcatg 2280
agtgtggagg taggacggct gcgtgccttc ttgcagggtg ggcaggaggc tacagatatt 2340
gccctcctgc tccgggatct ggaaacttca tgcagtgaca tccgccagtt ctgcaagaag 2400
atccgaaggc gaatgccagg gacagatgct cctgggatcc cagctgcact ggcctttgga 2460
ccacaggtat ctgacacgct cctagactgc aggaaacact tgacgtgggt cgtggctgtg 2520
ctgcaggagg tggcagctgc tgctgcccag ctcattgccc cactggcaga gaatgagggg 2580
ctacttgtgg ctgctctgga ggaactggct ttcaaagcaa gcgagcagat ctatgggacc 2640
ccctccagca gcccctatga gtgtctgcgc cagtcatgca acatcctcat cagtaccatg 2700
aacaagctgg ccacagccat gcaggagggg gagtatgatg cagagcggcc ccccagcaag 2760
cctccaccgg ttgaactgcg ggctgctgcc cttcgtgcag agatcacaga tgctgaaggc 2820
ctgggtttga agctcgaaga tcgagagaca gttattaagg agttgaagaa gtcactcaag 2880
attaagggag aggagctaag tgaggccaat gtgcggctga gcctcctgga gaagaagttg 2940
gacagtgctg ccaaggatgc agatgagcgc atcgagaaag tccagactcg gctggaggag 3000
acccaggcac tgctgcgaaa gaaggagaaa gagtttgagg agacaatgga tgcactccag 3060
gctgacatcg accagctgga ggcagagaag gcagaactaa agcagcgtct gaacagccag 3120
tccaaacgca cgattgaggg actccggggc cctcctcctt caggcattgc tactctggtc 3180
tctggcattg ctggtgaaga acagcagcga ggagccatcc ctgggcaggc tccagggtct 3240
gtgccaggcc cagggctggt gaaggactca ccactgctgc ttcagcagat ctctgccatg 3300
aggctgcaca tctcccagct ccagcatgag aacagcatcc tcaagggagc ccagatgaag 3360
gcatccttgg catccctgcc ccctctgcat gttgcaaagc tatcccatga gggccctggc 3420
agtgagttac cagctggagc gctgtatcgt aagaccagcc agctgctgga gacattgaat 3480
caattgagca cacacacgca cgtagtagac atcactcgca ccagccctgc tgccaagagc 3540
ccgtcggccc aacttatgga gcaagtggct cagcttaagt ccctgagtga caccgtcgag 3600
aagctcaagg atgaggtcct caaggagaca gtatctcagc gccctggagc cacagtaccc 3660
actgactttg ccaccttccc ttcatcagcc ttcctcaggg aggatccaaa gtgggaattc 3720
cctcggaaga acttggttct tggaaaaact ctaggagaag gcgaatttgg aaaagtggtc 3780
aaggcaacgg ccttccatct gaaaggcaga gcagggtaca ccacggtggc cgtgaagatg 3840
ctgaaagaga acgcctcccc gagtgagctt cgagacctgc tgtcagagtt caacgtcctg 3900
aagcaggtca accacccaca tgtcatcaaa ttgtatgggg cctgcagcca ggatggcccg 3960
ctcctcctca tcgtggagta cgccaaatac ggctccctgc ggggcttcct ccgcgagagc 4020
cgcaaagtgg ggcctggcta cctgggcagt ggaggcagcc gcaactccag ctccctggac 4080
cacccggatg agcgggccct caccatgggc gacctcatct catttgcctg gcagatctca 4140
caggggatgc agtatctggc cgagatgaag ctcgttcatc gggacttggc agccagaaac 4200
atcctggtag ctgaggggcg gaagatgaag atttcggatt tcggcttgtc ccgagatgtt 4260
tatgaagagg attcctacgt gaagaggagc cagggtcgga ttccagttaa atggatggca 4320
attgaatccc tttttgatca tatctacacc acgcaaagtg atgtatggtc ttttggtgtc 4380
ctgctgtggg agatcgtgac cctaggggga aacccctatc ctgggattcc tcctgagcgg 4440
ctcttcaacc ttctgaagac cggccaccgg atggagaggc cagacaactg cagcgaggag 4500
atgtaccgcc tgatgctgca atgctggaag caggagccgg acaaaaggcc ggtgtttgcg 4560
gacatcagca aagacctgga gaagatgatg gttaagagga gagactactt ggaccttgcg 4620
gcgtccactc catctgactc cctgatttat gacgacggcc tctcagagga ggagacaccg 4680
ctggtggact gtaataatgc ccccctccct cgagccctcc cttccacatg gattgaaaac 4740
aaactctatg gcatgtcaga cccgaactgg cctggagaga gtcctgtacc actcacgaga 4800
gctgatggca ctaacactgg gtttccaaga tatccaaatg atagtgtata tgctaactgg 4860
atgctttcac cctcagcggc aaaattaatg gacacgtttg atagttaa 4908
<210> 2
<211> 1635
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 2
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Arg Gly Leu Lys Pro Lys Lys Ala Pro Thr Ala Arg Lys
130 135 140
Thr Thr Thr Arg Arg Pro Lys Pro Thr Arg Pro Ala Ser Thr Gly Val
145 150 155 160
Ala Gly Ala Ser Ser Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly
165 170 175
Glu Leu Ser Ser Ser Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala
180 185 190
Ala Pro Ile Ile Pro Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro
195 200 205
Pro Leu Pro Ser Pro Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val
210 215 220
Arg Asp Leu Glu Glu Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu
225 230 235 240
Asp Lys Ala Lys Leu Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu
245 250 255
Gln Val Gln Glu Trp Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu
260 265 270
Gln Arg Arg Leu Lys Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu
275 280 285
Ala Lys Glu Arg Tyr Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile
290 295 300
Glu Met Ala Thr Leu Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser
305 310 315 320
Leu Gln Gln Glu Val Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr
325 330 335
Thr Asp Leu Glu Ile Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp
340 345 350
Gly Ala Ala Ser Ser Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala
355 360 365
Arg Leu Lys Asp Ala Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu
370 375 380
Lys Gln Glu His Val Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln
385 390 395 400
Glu Leu Glu Val Val Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu
405 410 415
Ser Gln Ala Glu Ser Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala
420 425 430
Ala Leu Gly Ala Glu Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu
435 440 445
Asn Leu Glu Glu Lys Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu
450 455 460
Glu Ala Met Asn Glu Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu
465 470 475 480
Thr Glu Leu Glu Leu Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val
485 490 495
Arg Glu Ala Gln Lys Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp
500 505 510
Tyr Gln Gln Thr Ile Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln
515 520 525
Asp Val Asn Arg Glu Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg
530 535 540
Gln Gln Gln Pro Pro Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala
545 550 555 560
Glu Thr Lys Ala His Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met
565 570 575
Glu Val Ala Gln Ala Asn Arg His Met Ser Leu Leu Thr Ala Phe Met
580 585 590
Pro Asp Ser Phe Leu Arg Pro Gly Gly Asp His Asp Cys Val Leu Val
595 600 605
Leu Leu Leu Met Pro Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys
610 615 620
Gln Ala Gln Glu Lys Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro
625 630 635 640
Gly Leu Arg Gly Ala Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu
645 650 655
Val Tyr Ser Leu Ser Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His
660 665 670
Ala Leu Ser Gln Cys Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu
675 680 685
Tyr Pro Glu Met Ser Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu
690 695 700
Leu Leu His Lys Asp Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu
705 710 715 720
Thr Lys Ala Ile Lys Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala
725 730 735
Glu Gln Pro Glu Asp Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe
740 745 750
Thr Gln Ser Ala Leu Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg
755 760 765
Ala Phe Leu Gln Gly Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu
770 775 780
Arg Asp Leu Glu Thr Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys
785 790 795 800
Ile Arg Arg Arg Met Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala
805 810 815
Leu Ala Phe Gly Pro Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys
820 825 830
His Leu Thr Trp Val Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala
835 840 845
Ala Gln Leu Ile Ala Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala
850 855 860
Ala Leu Glu Glu Leu Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr
865 870 875 880
Pro Ser Ser Ser Pro Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu
885 890 895
Ile Ser Thr Met Asn Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr
900 905 910
Asp Ala Glu Arg Pro Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala
915 920 925
Ala Ala Leu Arg Ala Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys
930 935 940
Leu Glu Asp Arg Glu Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys
945 950 955 960
Ile Lys Gly Glu Glu Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu
965 970 975
Glu Lys Lys Leu Asp Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu
980 985 990
Lys Val Gln Thr Arg Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys
995 1000 1005
Glu Lys Glu Phe Glu Glu Thr Met Asp Ala Leu Gln Ala Asp Ile
1010 1015 1020
Asp Gln Leu Glu Ala Glu Lys Ala Glu Leu Lys Gln Arg Leu Asn
1025 1030 1035
Ser Gln Ser Lys Arg Thr Ile Glu Gly Leu Arg Gly Pro Pro Pro
1040 1045 1050
Ser Gly Ile Ala Thr Leu Val Ser Gly Ile Ala Gly Glu Glu Gln
1055 1060 1065
Gln Arg Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser Val Pro Gly
1070 1075 1080
Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln Gln Ile Ser
1085 1090 1095
Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu Asn Ser Ile
1100 1105 1110
Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu Pro Pro
1115 1120 1125
Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu Leu
1130 1135 1140
Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu Thr
1145 1150 1155
Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp Ile Thr Arg
1160 1165 1170
Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu Met Glu Gln
1175 1180 1185
Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu Lys Leu Lys
1190 1195 1200
Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro Gly Ala Thr
1205 1210 1215
Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu Arg
1220 1225 1230
Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn Leu Val Leu Gly
1235 1240 1245
Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val Lys Ala Thr
1250 1255 1260
Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr Val Ala Val
1265 1270 1275
Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu Arg Asp Leu
1280 1285 1290
Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His Pro His Val
1295 1300 1305
Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu Leu Leu
1310 1315 1320
Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu Arg
1325 1330 1335
Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser
1340 1345 1350
Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr
1355 1360 1365
Met Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met
1370 1375 1380
Gln Tyr Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala Ala
1385 1390 1395
Arg Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp
1400 1405 1410
Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys
1415 1420 1425
Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala Ile Glu Ser
1430 1435 1440
Leu Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser Phe
1445 1450 1455
Gly Val Leu Leu Trp Glu Ile Val Thr Leu Gly Gly Asn Pro Tyr
1460 1465 1470
Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu Leu Lys Thr Gly
1475 1480 1485
His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu Met Tyr Arg
1490 1495 1500
Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys Arg Pro Val
1505 1510 1515
Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met Val Lys Arg
1520 1525 1530
Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro Ser Asp Ser Leu
1535 1540 1545
Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr Pro Leu Val Asp
1550 1555 1560
Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro Ser Thr Trp Ile
1565 1570 1575
Glu Asn Lys Leu Tyr Gly Met Ser Asp Pro Asn Trp Pro Gly Glu
1580 1585 1590
Ser Pro Val Pro Leu Thr Arg Ala Asp Gly Thr Asn Thr Gly Phe
1595 1600 1605
Pro Arg Tyr Pro Asn Asp Ser Val Tyr Ala Asn Trp Met Leu Ser
1610 1615 1620
Pro Ser Ala Ala Lys Leu Met Asp Thr Phe Asp Ser
1625 1630 1635
<210> 3
<211> 4782
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 3
atggcacaga gcaagaggca cgtgtacagc cggacgccca gcggcagcag gatgagtgcg 60
gaggcaagcg cccggcctct gcgggtgggc tcccgtgtag aggtgattgg aaaaggccac 120
cgaggcactg tggcctatgt tggagccaca ctgtttgcca ctggcaaatg ggtaggcgtg 180
attctggatg aagcaaaggg caaaaatgat ggaactgttc aaggcaggaa gtacttcact 240
tgtgatgaag ggcatggcat ctttgtgcgc cagtcccaga tccaggtatt tgaagatgga 300
gcagatacta cttccccaga gacacctgat tcttctgctt caaaagtcct caaaagagag 360
ggaactgata caactgcaaa gactagcaaa ctgcggggac tgaagcctaa gaaggcaccg 420
acagcccgaa agaccacaac tcggcgaccc aagcccacgc gcccagccag tactggggtg 480
gctggggcca gtagctccct gggcccctct ggctcagcgt cagcaggtga gctgagcagc 540
agtgagccca gcaccccggc tcagactccg ctggcagcac ccatcatccc cacgccggtc 600
ctcacctctc ctggagcagt ccccccgctt ccttccccat ccaaggagga ggagggacta 660
agggctcagg tgcgggacct ggaggagaaa ctagagaccc tgagactgaa acgggcagaa 720
gacaaagcaa agctaaaaga gctggagaaa cacaaaatcc agctggagca ggtgcaggaa 780
tggaagagca aaatgcagga gcagcaggcc gacctgcagc ggcgcctcaa ggaggcgaga 840
aaggaagcca aggaggcgct ggaggcaaag gaacgctata tggaggagat ggctgatact 900
gctgatgcca ttgagatggc cactttggac aaggagatgg ctgaagagcg ggctgagtcc 960
ctgcagcagg aggtggaggc actgaaggag cgggtggacg agctcactac tgacttagag 1020
atcctcaagg ctgagattga agagaagggc tcagatggcg ctgcatccag ttatcagctc 1080
aagcagcttg aggagcagaa tgcccgcctg aaggatgccc tggtgaggat gcgggatctt 1140
tcttcctcag agaagcagga gcatgtgaag ctccagaagc tcatggaaaa gaagaaccaa 1200
gagctggaag ttgtgaggca acagcgggag cgtctgcagg aggagctaag ccaggcagag 1260
agcaccattg atgagctcaa ggagcaggtg gatgctgctc tgggtgctga ggagatggtg 1320
gagatgctga cagatcggaa cctgaatctg gaagagaaag tgcgcgagtt gagggagact 1380
gtgggagact tggaagcgat gaatgagatg aacgatgagc tgcaggagaa tgcacgtgag 1440
acagaactgg agctgcggga gcagctggac atggcaggcg cgcgggttcg tgaggcccag 1500
aagcgtgtgg aggcagccca ggagacggtt gcagactacc agcagaccat caagaagtac 1560
cgccagctga ccgcccatct acaggatgtg aatcgggaac tgacaaacca gcaggaagca 1620
tctgtggaga ggcaacagca gccacctcca gagacctttg acttcaaaat caagtttgct 1680
gagactaagg cccatgccaa ggcaattgag atggaattga ggcagatgga ggtggcccag 1740
gccaatcgac acatgtccct gctgacagcc ttcatgcctg acagcttcct tcggccaggt 1800
ggggaccatg actgcgttct ggtgctgttg ctcatgcctc gtctcatttg caaggcagag 1860
ctgatccgga agcaggccca ggagaagttt gaactaagtg agaactgttc agagcggcct 1920
gggctgcgag gagctgctgg ggagcaactc agctttgctg ctggactggt gtactcgctg 1980
agcctgctgc aggccacgct acaccgctat gagcatgccc tctctcagtg cagtgtggat 2040
gtgtataaga aagtgggcag cctgtaccct gagatgagtg cccatgagcg ctccttggat 2100
ttcctcattg aactgctgca caaggatcag ctggatgaga ctgtcaatgt ggagcctctc 2160
accaaggcca tcaagtacta tcagcatctg tacagcatcc accttgccga acagcctgag 2220
gactgtacta tgcagctggc tgaccacatt aagttcacgc agagtgctct ggactgcatg 2280
agtgtggagg taggacggct gcgtgccttc ttgcagggtg ggcaggaggc tacagatatt 2340
gccctcctgc tccgggatct ggaaacttca tgcagtgaca tccgccagtt ctgcaagaag 2400
atccgaaggc gaatgccagg gacagatgct cctgggatcc cagctgcact ggcctttgga 2460
ccacaggtat ctgacacgct cctagactgc aggaaacact tgacgtgggt cgtggctgtg 2520
ctgcaggagg tggcagctgc tgctgcccag ctcattgccc cactggcaga gaatgagggg 2580
ctacttgtgg ctgctctgga ggaactggct ttcaaagcaa gcgagcagat ctatgggacc 2640
ccctccagca gcccctatga gtgtctgcgc cagtcatgca acatcctcat cagtaccatg 2700
aacaagctgg ccacagccat gcaggagggg gagtatgatg cagagcggcc ccccagcaag 2760
cctccaccgg ttgaactgcg ggctgctgcc cttcgtgcag agatcacaga tgctgaaggc 2820
ctgggtttga agctcgaaga tcgagagaca gttattaagg agttgaagaa gtcactcaag 2880
attaagggag aggagctaag tgaggccaat gtgcggctga gcctcctgga gaagaagttg 2940
gacagtgctg ccaaggatgc agatgagcgc atcgagaaag tccagactcg gctggaggag 3000
acccaggcac tgctgcgaaa gaaggagaaa gagtttgagg agacaatgga tgcactccag 3060
gctgacatcg accagctgga ggcagagaag gcagaactaa agcagcgtct gaacagccag 3120
tccaaacgca cgattgaggg actccggggc cctcctcctt caggcattgc tactctggtc 3180
tctggcattg ctggtgaaga acagcagcga ggagccatcc ctgggcaggc tccagggtct 3240
gtgccaggcc cagggctggt gaaggactca ccactgctgc ttcagcagat ctctgccatg 3300
aggctgcaca tctcccagct ccagcatgag aacagcatcc tcaagggagc ccagatgaag 3360
gcatccttgg catccctgcc ccctctgcat gttgcaaagc tatcccatga gggccctggc 3420
agtgagttac cagctggagc gctgtatcgt aagaccagcc agctgctgga gacattgaat 3480
caattgagca cacacacgca cgtagtagac atcactcgca ccagccctgc tgccaagagc 3540
ccgtcggccc aacttatgga gcaagtggct cagcttaagt ccctgagtga caccgtcgag 3600
aagctcaagg atgaggtcct caaggagaca gtatctcagc gccctggagc cacagtaccc 3660
actgactttg ccaccttccc ttcatcagcc ttcctcaggg aggatccaaa gtgggaattc 3720
cctcggaaga acttggttct tggaaaaact ctaggagaag gcgaatttgg aaaagtggtc 3780
aaggcaacgg ccttccatct gaaaggcaga gcagggtaca ccacggtggc cgtgaagatg 3840
ctgaaagaga acgcctcccc gagtgagctt cgagacctgc tgtcagagtt caacgtcctg 3900
aagcaggtca accacccaca tgtcatcaaa ttgtatgggg cctgcagcca ggatggcccg 3960
ctcctcctca tcgtggagta cgccaaatac ggctccctgc ggggcttcct ccgcgagagc 4020
cgcaaagtgg ggcctggcta cctgggcagt ggaggcagcc gcaactccag ctccctggac 4080
cacccggatg agcgggccct caccatgggc gacctcatct catttgcctg gcagatctca 4140
caggggatgc agtatctggc cgagatgaag ctcgttcatc gggacttggc agccagaaac 4200
atcctggtag ctgaggggcg gaagatgaag atttcggatt tcggcttgtc ccgagatgtt 4260
tatgaagagg attcctacgt gaagaggagc cagggtcgga ttccagttaa atggatggca 4320
attgaatccc tttttgatca tatctacacc acgcaaagtg atgtatggtc ttttggtgtc 4380
ctgctgtggg agatcgtgac cctaggggga aacccctatc ctgggattcc tcctgagcgg 4440
ctcttcaacc ttctgaagac cggccaccgg atggagaggc cagacaactg cagcgaggag 4500
atgtaccgcc tgatgctgca atgctggaag caggagccgg acaaaaggcc ggtgtttgcg 4560
gacatcagca aagacctgga gaagatgatg gttaagagga gagactactt ggaccttgcg 4620
gcgtccactc catctgactc cctgatttat gacgacggcc tctcagagga ggagacaccg 4680
ctggtggact gtaataatgc ccccctccct cgagccctcc cttccacatg gattgaaaac 4740
aaactctatg gtagaatttc ccatgcattt actagattct ag 4782
<210> 4
<211> 1593
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 4
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Arg Gly Leu Lys Pro Lys Lys Ala Pro Thr Ala Arg Lys
130 135 140
Thr Thr Thr Arg Arg Pro Lys Pro Thr Arg Pro Ala Ser Thr Gly Val
145 150 155 160
Ala Gly Ala Ser Ser Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly
165 170 175
Glu Leu Ser Ser Ser Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala
180 185 190
Ala Pro Ile Ile Pro Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro
195 200 205
Pro Leu Pro Ser Pro Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val
210 215 220
Arg Asp Leu Glu Glu Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu
225 230 235 240
Asp Lys Ala Lys Leu Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu
245 250 255
Gln Val Gln Glu Trp Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu
260 265 270
Gln Arg Arg Leu Lys Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu
275 280 285
Ala Lys Glu Arg Tyr Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile
290 295 300
Glu Met Ala Thr Leu Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser
305 310 315 320
Leu Gln Gln Glu Val Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr
325 330 335
Thr Asp Leu Glu Ile Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp
340 345 350
Gly Ala Ala Ser Ser Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala
355 360 365
Arg Leu Lys Asp Ala Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu
370 375 380
Lys Gln Glu His Val Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln
385 390 395 400
Glu Leu Glu Val Val Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu
405 410 415
Ser Gln Ala Glu Ser Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala
420 425 430
Ala Leu Gly Ala Glu Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu
435 440 445
Asn Leu Glu Glu Lys Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu
450 455 460
Glu Ala Met Asn Glu Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu
465 470 475 480
Thr Glu Leu Glu Leu Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val
485 490 495
Arg Glu Ala Gln Lys Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp
500 505 510
Tyr Gln Gln Thr Ile Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln
515 520 525
Asp Val Asn Arg Glu Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg
530 535 540
Gln Gln Gln Pro Pro Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala
545 550 555 560
Glu Thr Lys Ala His Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met
565 570 575
Glu Val Ala Gln Ala Asn Arg His Met Ser Leu Leu Thr Ala Phe Met
580 585 590
Pro Asp Ser Phe Leu Arg Pro Gly Gly Asp His Asp Cys Val Leu Val
595 600 605
Leu Leu Leu Met Pro Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys
610 615 620
Gln Ala Gln Glu Lys Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro
625 630 635 640
Gly Leu Arg Gly Ala Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu
645 650 655
Val Tyr Ser Leu Ser Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His
660 665 670
Ala Leu Ser Gln Cys Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu
675 680 685
Tyr Pro Glu Met Ser Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu
690 695 700
Leu Leu His Lys Asp Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu
705 710 715 720
Thr Lys Ala Ile Lys Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala
725 730 735
Glu Gln Pro Glu Asp Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe
740 745 750
Thr Gln Ser Ala Leu Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg
755 760 765
Ala Phe Leu Gln Gly Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu
770 775 780
Arg Asp Leu Glu Thr Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys
785 790 795 800
Ile Arg Arg Arg Met Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala
805 810 815
Leu Ala Phe Gly Pro Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys
820 825 830
His Leu Thr Trp Val Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala
835 840 845
Ala Gln Leu Ile Ala Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala
850 855 860
Ala Leu Glu Glu Leu Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr
865 870 875 880
Pro Ser Ser Ser Pro Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu
885 890 895
Ile Ser Thr Met Asn Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr
900 905 910
Asp Ala Glu Arg Pro Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala
915 920 925
Ala Ala Leu Arg Ala Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys
930 935 940
Leu Glu Asp Arg Glu Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys
945 950 955 960
Ile Lys Gly Glu Glu Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu
965 970 975
Glu Lys Lys Leu Asp Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu
980 985 990
Lys Val Gln Thr Arg Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys
995 1000 1005
Glu Lys Glu Phe Glu Glu Thr Met Asp Ala Leu Gln Ala Asp Ile
1010 1015 1020
Asp Gln Leu Glu Ala Glu Lys Ala Glu Leu Lys Gln Arg Leu Asn
1025 1030 1035
Ser Gln Ser Lys Arg Thr Ile Glu Gly Leu Arg Gly Pro Pro Pro
1040 1045 1050
Ser Gly Ile Ala Thr Leu Val Ser Gly Ile Ala Gly Glu Glu Gln
1055 1060 1065
Gln Arg Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser Val Pro Gly
1070 1075 1080
Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln Gln Ile Ser
1085 1090 1095
Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu Asn Ser Ile
1100 1105 1110
Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu Pro Pro
1115 1120 1125
Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu Leu
1130 1135 1140
Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu Thr
1145 1150 1155
Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp Ile Thr Arg
1160 1165 1170
Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu Met Glu Gln
1175 1180 1185
Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu Lys Leu Lys
1190 1195 1200
Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro Gly Ala Thr
1205 1210 1215
Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu Arg
1220 1225 1230
Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn Leu Val Leu Gly
1235 1240 1245
Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val Lys Ala Thr
1250 1255 1260
Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr Val Ala Val
1265 1270 1275
Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu Arg Asp Leu
1280 1285 1290
Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His Pro His Val
1295 1300 1305
Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu Leu Leu
1310 1315 1320
Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu Arg
1325 1330 1335
Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser
1340 1345 1350
Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr
1355 1360 1365
Met Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met
1370 1375 1380
Gln Tyr Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala Ala
1385 1390 1395
Arg Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp
1400 1405 1410
Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys
1415 1420 1425
Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala Ile Glu Ser
1430 1435 1440
Leu Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser Phe
1445 1450 1455
Gly Val Leu Leu Trp Glu Ile Val Thr Leu Gly Gly Asn Pro Tyr
1460 1465 1470
Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu Leu Lys Thr Gly
1475 1480 1485
His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu Met Tyr Arg
1490 1495 1500
Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys Arg Pro Val
1505 1510 1515
Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met Val Lys Arg
1520 1525 1530
Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro Ser Asp Ser Leu
1535 1540 1545
Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr Pro Leu Val Asp
1550 1555 1560
Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro Ser Thr Trp Ile
1565 1570 1575
Glu Asn Lys Leu Tyr Gly Arg Ile Ser His Ala Phe Thr Arg Phe
1580 1585 1590
<210> 5
<211> 4506
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 5
atgatgagac aggcaccgac agcccgaaag accacaactc ggcgacccaa gcccacgcgc 60
ccagccagta ctggggtggc tggggccagt agctccctgg gcccctctgg ctcagcgtca 120
gcaggtgagc tgagcagcag tgagcccagc accccggctc agactccgct ggcagcaccc 180
atcatcccca cgccggtcct cacctctcct ggagcagtcc ccccgcttcc ttccccatcc 240
aaggaggagg agggactaag ggctcaggtg cgggacctgg aggagaaact agagaccctg 300
agactgaaac gggcagaaga caaagcaaag ctaaaagagc tggagaaaca caaaatccag 360
ctggagcagg tgcaggaatg gaagagcaaa atgcaggagc agcaggccga cctgcagcgg 420
cgcctcaagg aggcgagaaa ggaagccaag gaggcgctgg aggcaaagga acgctatatg 480
gaggagatgg ctgatactgc tgatgccatt gagatggcca ctttggacaa ggagatggct 540
gaagagcggg ctgagtccct gcagcaggag gtggaggcac tgaaggagcg ggtggacgag 600
ctcactactg acttagagat cctcaaggct gagattgaag agaagggctc agatggcgct 660
gcatccagtt atcagctcaa gcagcttgag gagcagaatg cccgcctgaa ggatgccctg 720
gtgaggatgc gggatctttc ttcctcagag aagcaggagc atgtgaagct ccagaagctc 780
atggaaaaga agaaccaaga gctggaagtt gtgaggcaac agcgggagcg tctgcaggag 840
gagctaagcc aggcagagag caccattgat gagctcaagg agcaggtgga tgctgctctg 900
ggtgctgagg agatggtgga gatgctgaca gatcggaacc tgaatctgga agagaaagtg 960
cgcgagttga gggagactgt gggagacttg gaagcgatga atgagatgaa cgatgagctg 1020
caggagaatg cacgtgagac agaactggag ctgcgggagc agctggacat ggcaggcgcg 1080
cgggttcgtg aggcccagaa gcgtgtggag gcagcccagg agacggttgc agactaccag 1140
cagaccatca agaagtaccg ccagctgacc gcccatctac aggatgtgaa tcgggaactg 1200
acaaaccagc aggaagcatc tgtggagagg caacagcagc cacctccaga gacctttgac 1260
ttcaaaatca agtttgctga gactaaggcc catgccaagg caattgagat ggaattgagg 1320
cagatggagg tggcccaggc caatcgacac atgtccctgc tgacagcctt catgcctgac 1380
agcttccttc ggccaggtgg ggaccatgac tgcgttctgg tgctgttgct catgcctcgt 1440
ctcatttgca aggcagagct gatccggaag caggcccagg agaagtttga actaagtgag 1500
aactgttcag agcggcctgg gctgcgagga gctgctgggg agcaactcag ctttgctgct 1560
ggactggtgt actcgctgag cctgctgcag gccacgctac accgctatga gcatgccctc 1620
tctcagtgca gtgtggatgt gtataagaaa gtgggcagcc tgtaccctga gatgagtgcc 1680
catgagcgct ccttggattt cctcattgaa ctgctgcaca aggatcagct ggatgagact 1740
gtcaatgtgg agcctctcac caaggccatc aagtactatc agcatctgta cagcatccac 1800
cttgccgaac agcctgagga ctgtactatg cagctggctg accacattaa gttcacgcag 1860
agtgctctgg actgcatgag tgtggaggta ggacggctgc gtgccttctt gcagggtggg 1920
caggaggcta cagatattgc cctcctgctc cgggatctgg aaacttcatg cagtgacatc 1980
cgccagttct gcaagaagat ccgaaggcga atgccaggga cagatgctcc tgggatccca 2040
gctgcactgg cctttggacc acaggtatct gacacgctcc tagactgcag gaaacacttg 2100
acgtgggtcg tggctgtgct gcaggaggtg gcagctgctg ctgcccagct cattgcccca 2160
ctggcagaga atgaggggct acttgtggct gctctggagg aactggcttt caaagcaagc 2220
gagcagatct atgggacccc ctccagcagc ccctatgagt gtctgcgcca gtcatgcaac 2280
atcctcatca gtaccatgaa caagctggcc acagccatgc aggaggggga gtatgatgca 2340
gagcggcccc ccagcaagcc tccaccggtt gaactgcggg ctgctgccct tcgtgcagag 2400
atcacagatg ctgaaggcct gggtttgaag ctcgaagatc gagagacagt tattaaggag 2460
ttgaagaagt cactcaagat taagggagag gagctaagtg aggccaatgt gcggctgagc 2520
ctcctggaga agaagttgga cagtgctgcc aaggatgcag atgagcgcat cgagaaagtc 2580
cagactcggc tggaggagac ccaggcactg ctgcgaaaga aggagaaaga gtttgaggag 2640
acaatggatg cactccaggc tgacatcgac cagctggagg cagagaaggc agaactaaag 2700
cagcgtctga acagccagtc caaacgcacg attgagggac tccggggccc tcctccttca 2760
ggcattgcta ctctggtctc tggcattgct ggtgaagaac agcagcgagg agccatccct 2820
gggcaggctc cagggtctgt gccaggccca gggctggtga aggactcacc actgctgctt 2880
cagcagatct ctgccatgag gctgcacatc tcccagctcc agcatgagaa cagcatcctc 2940
aagggagccc agatgaaggc atccttggca tccctgcccc ctctgcatgt tgcaaagcta 3000
tcccatgagg gccctggcag tgagttacca gctggagcgc tgtatcgtaa gaccagccag 3060
ctgctggaga cattgaatca attgagcaca cacacgcacg tagtagacat cactcgcacc 3120
agccctgctg ccaagagccc gtcggcccaa cttatggagc aagtggctca gcttaagtcc 3180
ctgagtgaca ccgtcgagaa gctcaaggat gaggtcctca aggagacagt atctcagcgc 3240
cctggagcca cagtacccac tgactttgcc accttccctt catcagcctt cctcagggag 3300
gatccaaagt gggaattccc tcggaagaac ttggttcttg gaaaaactct aggagaaggc 3360
gaatttggaa aagtggtcaa ggcaacggcc ttccatctga aaggcagagc agggtacacc 3420
acggtggccg tgaagatgct gaaagagaac gcctccccga gtgagcttcg agacctgctg 3480
tcagagttca acgtcctgaa gcaggtcaac cacccacatg tcatcaaatt gtatggggcc 3540
tgcagccagg atggcccgct cctcctcatc gtggagtacg ccaaatacgg ctccctgcgg 3600
ggcttcctcc gcgagagccg caaagtgggg cctggctacc tgggcagtgg aggcagccgc 3660
aactccagct ccctggacca cccggatgag cgggccctca ccatgggcga cctcatctca 3720
tttgcctggc agatctcaca ggggatgcag tatctggccg agatgaagct cgttcatcgg 3780
gacttggcag ccagaaacat cctggtagct gaggggcgga agatgaagat ttcggatttc 3840
ggcttgtccc gagatgttta tgaagaggat tcctacgtga agaggagcca gggtcggatt 3900
ccagttaaat ggatggcaat tgaatccctt tttgatcata tctacaccac gcaaagtgat 3960
gtatggtctt ttggtgtcct gctgtgggag atcgtgaccc tagggggaaa cccctatcct 4020
gggattcctc ctgagcggct cttcaacctt ctgaagaccg gccaccggat ggagaggcca 4080
gacaactgca gcgaggagat gtaccgcctg atgctgcaat gctggaagca ggagccggac 4140
aaaaggccgg tgtttgcgga catcagcaaa gacctggaga agatgatggt taagaggaga 4200
gactacttgg accttgcggc gtccactcca tctgactccc tgatttatga cgacggcctc 4260
tcagaggagg agacaccgct ggtggactgt aataatgccc ccctccctcg agccctccct 4320
tccacatgga ttgaaaacaa actctatggc atgtcagacc cgaactggcc tggagagagt 4380
cctgtaccac tcacgagagc tgatggcact aacactgggt ttccaagata tccaaatgat 4440
agtgtatatg ctaactggat gctttcaccc tcagcggcaa aattaatgga cacgtttgat 4500
agttaa 4506
<210> 6
<211> 1501
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 6
Met Met Arg Gln Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro
1 5 10 15
Lys Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser
20 25 30
Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu
35 40 45
Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr
50 55 60
Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser
65 70 75 80
Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys
85 90 95
Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys
100 105 110
Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys
115 120 125
Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu
130 135 140
Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met
145 150 155 160
Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp
165 170 175
Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu
180 185 190
Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu
195 200 205
Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr
210 215 220
Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu
225 230 235 240
Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys
245 250 255
Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg
260 265 270
Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr
275 280 285
Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu
290 295 300
Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val
305 310 315 320
Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met
325 330 335
Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg
340 345 350
Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg
355 360 365
Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys
370 375 380
Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu
385 390 395 400
Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro
405 410 415
Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala
420 425 430
Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn
435 440 445
Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg
450 455 460
Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg
465 470 475 480
Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe
485 490 495
Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala
500 505 510
Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu
515 520 525
Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser
530 535 540
Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala
545 550 555 560
His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln
565 570 575
Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr
580 585 590
Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys
595 600 605
Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp
610 615 620
Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly
625 630 635 640
Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser
645 650 655
Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro
660 665 670
Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln
675 680 685
Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val
690 695 700
Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro
705 710 715 720
Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala
725 730 735
Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr
740 745 750
Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys
755 760 765
Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro
770 775 780
Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu
785 790 795 800
Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr
805 810 815
Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu
820 825 830
Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser
835 840 845
Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu
850 855 860
Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu
865 870 875 880
Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
885 890 895
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile Glu
900 905 910
Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val Ser Gly
915 920 925
Ile Ala Gly Glu Glu Gln Gln Arg Gly Ala Ile Pro Gly Gln Ala Pro
930 935 940
Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu
945 950 955 960
Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu
965 970 975
Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu
980 985 990
Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu
995 1000 1005
Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu
1010 1015 1020
Thr Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp Ile Thr
1025 1030 1035
Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu Met Glu
1040 1045 1050
Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu Lys Leu
1055 1060 1065
Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro Gly Ala
1070 1075 1080
Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu
1085 1090 1095
Arg Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn Leu Val Leu
1100 1105 1110
Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val Lys Ala
1115 1120 1125
Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr Val Ala
1130 1135 1140
Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu Arg Asp
1145 1150 1155
Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His Pro His
1160 1165 1170
Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu Leu
1175 1180 1185
Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu
1190 1195 1200
Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly
1205 1210 1215
Ser Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu
1220 1225 1230
Thr Met Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly
1235 1240 1245
Met Gln Tyr Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala
1250 1255 1260
Ala Arg Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser
1265 1270 1275
Asp Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val
1280 1285 1290
Lys Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala Ile Glu
1295 1300 1305
Ser Leu Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser
1310 1315 1320
Phe Gly Val Leu Leu Trp Glu Ile Val Thr Leu Gly Gly Asn Pro
1325 1330 1335
Tyr Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu Leu Lys Thr
1340 1345 1350
Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu Met Tyr
1355 1360 1365
Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys Arg Pro
1370 1375 1380
Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met Val Lys
1385 1390 1395
Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro Ser Asp Ser
1400 1405 1410
Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr Pro Leu Val
1415 1420 1425
Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro Ser Thr Trp
1430 1435 1440
Ile Glu Asn Lys Leu Tyr Gly Met Ser Asp Pro Asn Trp Pro Gly
1445 1450 1455
Glu Ser Pro Val Pro Leu Thr Arg Ala Asp Gly Thr Asn Thr Gly
1460 1465 1470
Phe Pro Arg Tyr Pro Asn Asp Ser Val Tyr Ala Asn Trp Met Leu
1475 1480 1485
Ser Pro Ser Ala Ala Lys Leu Met Asp Thr Phe Asp Ser
1490 1495 1500
<210> 7
<211> 4380
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 7
atgatgagac aggcaccgac agcccgaaag accacaactc ggcgacccaa gcccacgcgc 60
ccagccagta ctggggtggc tggggccagt agctccctgg gcccctctgg ctcagcgtca 120
gcaggtgagc tgagcagcag tgagcccagc accccggctc agactccgct ggcagcaccc 180
atcatcccca cgccggtcct cacctctcct ggagcagtcc ccccgcttcc ttccccatcc 240
aaggaggagg agggactaag ggctcaggtg cgggacctgg aggagaaact agagaccctg 300
agactgaaac gggcagaaga caaagcaaag ctaaaagagc tggagaaaca caaaatccag 360
ctggagcagg tgcaggaatg gaagagcaaa atgcaggagc agcaggccga cctgcagcgg 420
cgcctcaagg aggcgagaaa ggaagccaag gaggcgctgg aggcaaagga acgctatatg 480
gaggagatgg ctgatactgc tgatgccatt gagatggcca ctttggacaa ggagatggct 540
gaagagcggg ctgagtccct gcagcaggag gtggaggcac tgaaggagcg ggtggacgag 600
ctcactactg acttagagat cctcaaggct gagattgaag agaagggctc agatggcgct 660
gcatccagtt atcagctcaa gcagcttgag gagcagaatg cccgcctgaa ggatgccctg 720
gtgaggatgc gggatctttc ttcctcagag aagcaggagc atgtgaagct ccagaagctc 780
atggaaaaga agaaccaaga gctggaagtt gtgaggcaac agcgggagcg tctgcaggag 840
gagctaagcc aggcagagag caccattgat gagctcaagg agcaggtgga tgctgctctg 900
ggtgctgagg agatggtgga gatgctgaca gatcggaacc tgaatctgga agagaaagtg 960
cgcgagttga gggagactgt gggagacttg gaagcgatga atgagatgaa cgatgagctg 1020
caggagaatg cacgtgagac agaactggag ctgcgggagc agctggacat ggcaggcgcg 1080
cgggttcgtg aggcccagaa gcgtgtggag gcagcccagg agacggttgc agactaccag 1140
cagaccatca agaagtaccg ccagctgacc gcccatctac aggatgtgaa tcgggaactg 1200
acaaaccagc aggaagcatc tgtggagagg caacagcagc cacctccaga gacctttgac 1260
ttcaaaatca agtttgctga gactaaggcc catgccaagg caattgagat ggaattgagg 1320
cagatggagg tggcccaggc caatcgacac atgtccctgc tgacagcctt catgcctgac 1380
agcttccttc ggccaggtgg ggaccatgac tgcgttctgg tgctgttgct catgcctcgt 1440
ctcatttgca aggcagagct gatccggaag caggcccagg agaagtttga actaagtgag 1500
aactgttcag agcggcctgg gctgcgagga gctgctgggg agcaactcag ctttgctgct 1560
ggactggtgt actcgctgag cctgctgcag gccacgctac accgctatga gcatgccctc 1620
tctcagtgca gtgtggatgt gtataagaaa gtgggcagcc tgtaccctga gatgagtgcc 1680
catgagcgct ccttggattt cctcattgaa ctgctgcaca aggatcagct ggatgagact 1740
gtcaatgtgg agcctctcac caaggccatc aagtactatc agcatctgta cagcatccac 1800
cttgccgaac agcctgagga ctgtactatg cagctggctg accacattaa gttcacgcag 1860
agtgctctgg actgcatgag tgtggaggta ggacggctgc gtgccttctt gcagggtggg 1920
caggaggcta cagatattgc cctcctgctc cgggatctgg aaacttcatg cagtgacatc 1980
cgccagttct gcaagaagat ccgaaggcga atgccaggga cagatgctcc tgggatccca 2040
gctgcactgg cctttggacc acaggtatct gacacgctcc tagactgcag gaaacacttg 2100
acgtgggtcg tggctgtgct gcaggaggtg gcagctgctg ctgcccagct cattgcccca 2160
ctggcagaga atgaggggct acttgtggct gctctggagg aactggcttt caaagcaagc 2220
gagcagatct atgggacccc ctccagcagc ccctatgagt gtctgcgcca gtcatgcaac 2280
atcctcatca gtaccatgaa caagctggcc acagccatgc aggaggggga gtatgatgca 2340
gagcggcccc ccagcaagcc tccaccggtt gaactgcggg ctgctgccct tcgtgcagag 2400
atcacagatg ctgaaggcct gggtttgaag ctcgaagatc gagagacagt tattaaggag 2460
ttgaagaagt cactcaagat taagggagag gagctaagtg aggccaatgt gcggctgagc 2520
ctcctggaga agaagttgga cagtgctgcc aaggatgcag atgagcgcat cgagaaagtc 2580
cagactcggc tggaggagac ccaggcactg ctgcgaaaga aggagaaaga gtttgaggag 2640
acaatggatg cactccaggc tgacatcgac cagctggagg cagagaaggc agaactaaag 2700
cagcgtctga acagccagtc caaacgcacg attgagggac tccggggccc tcctccttca 2760
ggcattgcta ctctggtctc tggcattgct ggtgaagaac agcagcgagg agccatccct 2820
gggcaggctc cagggtctgt gccaggccca gggctggtga aggactcacc actgctgctt 2880
cagcagatct ctgccatgag gctgcacatc tcccagctcc agcatgagaa cagcatcctc 2940
aagggagccc agatgaaggc atccttggca tccctgcccc ctctgcatgt tgcaaagcta 3000
tcccatgagg gccctggcag tgagttacca gctggagcgc tgtatcgtaa gaccagccag 3060
ctgctggaga cattgaatca attgagcaca cacacgcacg tagtagacat cactcgcacc 3120
agccctgctg ccaagagccc gtcggcccaa cttatggagc aagtggctca gcttaagtcc 3180
ctgagtgaca ccgtcgagaa gctcaaggat gaggtcctca aggagacagt atctcagcgc 3240
cctggagcca cagtacccac tgactttgcc accttccctt catcagcctt cctcagggag 3300
gatccaaagt gggaattccc tcggaagaac ttggttcttg gaaaaactct aggagaaggc 3360
gaatttggaa aagtggtcaa ggcaacggcc ttccatctga aaggcagagc agggtacacc 3420
acggtggccg tgaagatgct gaaagagaac gcctccccga gtgagcttcg agacctgctg 3480
tcagagttca acgtcctgaa gcaggtcaac cacccacatg tcatcaaatt gtatggggcc 3540
tgcagccagg atggcccgct cctcctcatc gtggagtacg ccaaatacgg ctccctgcgg 3600
ggcttcctcc gcgagagccg caaagtgggg cctggctacc tgggcagtgg aggcagccgc 3660
aactccagct ccctggacca cccggatgag cgggccctca ccatgggcga cctcatctca 3720
tttgcctggc agatctcaca ggggatgcag tatctggccg agatgaagct cgttcatcgg 3780
gacttggcag ccagaaacat cctggtagct gaggggcgga agatgaagat ttcggatttc 3840
ggcttgtccc gagatgttta tgaagaggat tcctacgtga agaggagcca gggtcggatt 3900
ccagttaaat ggatggcaat tgaatccctt tttgatcata tctacaccac gcaaagtgat 3960
gtatggtctt ttggtgtcct gctgtgggag atcgtgaccc tagggggaaa cccctatcct 4020
gggattcctc ctgagcggct cttcaacctt ctgaagaccg gccaccggat ggagaggcca 4080
gacaactgca gcgaggagat gtaccgcctg atgctgcaat gctggaagca ggagccggac 4140
aaaaggccgg tgtttgcgga catcagcaaa gacctggaga agatgatggt taagaggaga 4200
gactacttgg accttgcggc gtccactcca tctgactccc tgatttatga cgacggcctc 4260
tcagaggagg agacaccgct ggtggactgt aataatgccc ccctccctcg agccctccct 4320
tccacatgga ttgaaaacaa actctatggt agaatttccc atgcatttac tagattctag 4380
<210> 8
<211> 1459
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 8
Met Met Arg Gln Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro
1 5 10 15
Lys Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser
20 25 30
Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu
35 40 45
Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr
50 55 60
Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser
65 70 75 80
Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys
85 90 95
Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys
100 105 110
Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys
115 120 125
Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu
130 135 140
Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met
145 150 155 160
Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp
165 170 175
Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu
180 185 190
Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu
195 200 205
Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr
210 215 220
Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu
225 230 235 240
Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys
245 250 255
Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg
260 265 270
Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr
275 280 285
Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu
290 295 300
Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val
305 310 315 320
Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met
325 330 335
Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg
340 345 350
Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg
355 360 365
Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys
370 375 380
Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu
385 390 395 400
Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro
405 410 415
Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala
420 425 430
Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn
435 440 445
Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg
450 455 460
Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg
465 470 475 480
Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe
485 490 495
Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala
500 505 510
Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu
515 520 525
Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser
530 535 540
Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala
545 550 555 560
His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln
565 570 575
Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr
580 585 590
Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys
595 600 605
Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp
610 615 620
Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly
625 630 635 640
Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser
645 650 655
Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro
660 665 670
Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln
675 680 685
Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val
690 695 700
Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro
705 710 715 720
Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala
725 730 735
Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr
740 745 750
Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys
755 760 765
Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro
770 775 780
Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu
785 790 795 800
Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr
805 810 815
Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu
820 825 830
Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser
835 840 845
Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu
850 855 860
Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu
865 870 875 880
Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
885 890 895
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile Glu
900 905 910
Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val Ser Gly
915 920 925
Ile Ala Gly Glu Glu Gln Gln Arg Gly Ala Ile Pro Gly Gln Ala Pro
930 935 940
Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu
945 950 955 960
Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu
965 970 975
Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu
980 985 990
Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu
995 1000 1005
Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu
1010 1015 1020
Thr Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp Ile Thr
1025 1030 1035
Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu Met Glu
1040 1045 1050
Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu Lys Leu
1055 1060 1065
Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro Gly Ala
1070 1075 1080
Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu
1085 1090 1095
Arg Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn Leu Val Leu
1100 1105 1110
Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val Lys Ala
1115 1120 1125
Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr Val Ala
1130 1135 1140
Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu Arg Asp
1145 1150 1155
Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His Pro His
1160 1165 1170
Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu Leu
1175 1180 1185
Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu
1190 1195 1200
Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly
1205 1210 1215
Ser Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu
1220 1225 1230
Thr Met Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly
1235 1240 1245
Met Gln Tyr Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala
1250 1255 1260
Ala Arg Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser
1265 1270 1275
Asp Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val
1280 1285 1290
Lys Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala Ile Glu
1295 1300 1305
Ser Leu Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser
1310 1315 1320
Phe Gly Val Leu Leu Trp Glu Ile Val Thr Leu Gly Gly Asn Pro
1325 1330 1335
Tyr Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu Leu Lys Thr
1340 1345 1350
Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu Met Tyr
1355 1360 1365
Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys Arg Pro
1370 1375 1380
Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met Val Lys
1385 1390 1395
Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro Ser Asp Ser
1400 1405 1410
Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr Pro Leu Val
1415 1420 1425
Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro Ser Thr Trp
1430 1435 1440
Ile Glu Asn Lys Leu Tyr Gly Arg Ile Ser His Ala Phe Thr Arg
1445 1450 1455
Phe
<210> 9
<211> 4833
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 9
atggcacaga gcaagaggca cgtgtacagc cggacgccca gcggcagcag gatgagtgcg 60
gaggcaagcg cccggcctct gcgggtgggc tcccgtgtag aggtgattgg aaaaggccac 120
cgaggcactg tggcctatgt tggagccaca ctgtttgcca ctggcaaatg ggtaggcgtg 180
attctggatg aagcaaaggg caaaaatgat ggaactgttc aaggcaggaa gtacttcact 240
tgtgatgaag ggcatggcat ctttgtgcgc cagtcccaga tccaggtatt tgaagatgga 300
gcagatacta cttccccaga gacacctgat tcttctgctt caaaagtcct caaaagagag 360
ggaactgata caactgcaaa gactagcaaa ctgcccacgc gcccagccag tactggggtg 420
gctggggcca gtagctccct gggcccctct ggctcagcgt cagcaggtga gctgagcagc 480
agtgagccca gcaccccggc tcagactccg ctggcagcac ccatcatccc cacgccggtc 540
ctcacctctc ctggagcagt ccccccgctt ccttccccat ccaaggagga ggagggacta 600
agggctcagg tgcgggacct ggaggagaaa ctagagaccc tgagactgaa acgggcagaa 660
gacaaagcaa agctaaaaga gctggagaaa cacaaaatcc agctggagca ggtgcaggaa 720
tggaagagca aaatgcagga gcagcaggcc gacctgcagc ggcgcctcaa ggaggcgaga 780
aaggaagcca aggaggcgct ggaggcaaag gaacgctata tggaggagat ggctgatact 840
gctgatgcca ttgagatggc cactttggac aaggagatgg ctgaagagcg ggctgagtcc 900
ctgcagcagg aggtggaggc actgaaggag cgggtggacg agctcactac tgacttagag 960
atcctcaagg ctgagattga agagaagggc tcagatggcg ctgcatccag ttatcagctc 1020
aagcagcttg aggagcagaa tgcccgcctg aaggatgccc tggtgaggat gcgggatctt 1080
tcttcctcag agaagcagga gcatgtgaag ctccagaagc tcatggaaaa gaagaaccaa 1140
gagctggaag ttgtgaggca acagcgggag cgtctgcagg aggagctaag ccaggcagag 1200
agcaccattg atgagctcaa ggagcaggtg gatgctgctc tgggtgctga ggagatggtg 1260
gagatgctga cagatcggaa cctgaatctg gaagagaaag tgcgcgagtt gagggagact 1320
gtgggagact tggaagcgat gaatgagatg aacgatgagc tgcaggagaa tgcacgtgag 1380
acagaactgg agctgcggga gcagctggac atggcaggcg cgcgggttcg tgaggcccag 1440
aagcgtgtgg aggcagccca ggagacggtt gcagactacc agcagaccat caagaagtac 1500
cgccagctga ccgcccatct acaggatgtg aatcgggaac tgacaaacca gcaggaagca 1560
tctgtggaga ggcaacagca gccacctcca gagacctttg acttcaaaat caagtttgct 1620
gagactaagg cccatgccaa ggcaattgag atggaattga ggcagatgga ggtggcccag 1680
gccaatcgac acatgtccct gctgacagcc ttcatgcctg acagcttcct tcggccaggt 1740
ggggaccatg actgcgttct ggtgctgttg ctcatgcctc gtctcatttg caaggcagag 1800
ctgatccgga agcaggccca ggagaagttt gaactaagtg agaactgttc agagcggcct 1860
gggctgcgag gagctgctgg ggagcaactc agctttgctg ctggactggt gtactcgctg 1920
agcctgctgc aggccacgct acaccgctat gagcatgccc tctctcagtg cagtgtggat 1980
gtgtataaga aagtgggcag cctgtaccct gagatgagtg cccatgagcg ctccttggat 2040
ttcctcattg aactgctgca caaggatcag ctggatgaga ctgtcaatgt ggagcctctc 2100
accaaggcca tcaagtacta tcagcatctg tacagcatcc accttgccga acagcctgag 2160
gactgtacta tgcagctggc tgaccacatt aagttcacgc agagtgctct ggactgcatg 2220
agtgtggagg taggacggct gcgtgccttc ttgcagggtg ggcaggaggc tacagatatt 2280
gccctcctgc tccgggatct ggaaacttca tgcagtgaca tccgccagtt ctgcaagaag 2340
atccgaaggc gaatgccagg gacagatgct cctgggatcc cagctgcact ggcctttgga 2400
ccacaggtat ctgacacgct cctagactgc aggaaacact tgacgtgggt cgtggctgtg 2460
ctgcaggagg tggcagctgc tgctgcccag ctcattgccc cactggcaga gaatgagggg 2520
ctacttgtgg ctgctctgga ggaactggct ttcaaagcaa gcgagcagat ctatgggacc 2580
ccctccagca gcccctatga gtgtctgcgc cagtcatgca acatcctcat cagtaccatg 2640
aacaagctgg ccacagccat gcaggagggg gagtatgatg cagagcggcc ccccagcaag 2700
cctccaccgg ttgaactgcg ggctgctgcc cttcgtgcag agatcacaga tgctgaaggc 2760
ctgggtttga agctcgaaga tcgagagaca gttattaagg agttgaagaa gtcactcaag 2820
attaagggag aggagctaag tgaggccaat gtgcggctga gcctcctgga gaagaagttg 2880
gacagtgctg ccaaggatgc agatgagcgc atcgagaaag tccagactcg gctggaggag 2940
acccaggcac tgctgcgaaa gaaggagaaa gagtttgagg agacaatgga tgcactccag 3000
gctgacatcg accagctgga ggcagagaag gcagaactaa agcagcgtct gaacagccag 3060
tccaaacgca cgattgaggg actccggggc cctcctcctt caggcattgc tactctggtc 3120
tctggcattg ctggtggagc catccctggg caggctccag ggtctgtgcc aggcccaggg 3180
ctggtgaagg actcaccact gctgcttcag cagatctctg ccatgaggct gcacatctcc 3240
cagctccagc atgagaacag catcctcaag ggagcccaga tgaaggcatc cttggcatcc 3300
ctgccccctc tgcatgttgc aaagctatcc catgagggcc ctggcagtga gttaccagct 3360
ggagcgctgt atcgtaagac cagccagctg ctggagacat tgaatcaatt gagcacacac 3420
acgcacgtag tagacatcac tcgcaccagc cctgctgcca agagcccgtc ggcccaactt 3480
atggagcaag tggctcagct taagtccctg agtgacaccg tcgagaagct caaggatgag 3540
gtcctcaagg agacagtatc tcagcgccct ggagccacag tacccactga ctttgccacc 3600
ttcccttcat cagccttcct cagggaggat ccaaagtggg aattccctcg gaagaacttg 3660
gttcttggaa aaactctagg agaaggcgaa tttggaaaag tggtcaaggc aacggccttc 3720
catctgaaag gcagagcagg gtacaccacg gtggccgtga agatgctgaa agagaacgcc 3780
tccccgagtg agcttcgaga cctgctgtca gagttcaacg tcctgaagca ggtcaaccac 3840
ccacatgtca tcaaattgta tggggcctgc agccaggatg gcccgctcct cctcatcgtg 3900
gagtacgcca aatacggctc cctgcggggc ttcctccgcg agagccgcaa agtggggcct 3960
ggctacctgg gcagtggagg cagccgcaac tccagctccc tggaccaccc ggatgagcgg 4020
gccctcacca tgggcgacct catctcattt gcctggcaga tctcacaggg gatgcagtat 4080
ctggccgaga tgaagctcgt tcatcgggac ttggcagcca gaaacatcct ggtagctgag 4140
gggcggaaga tgaagatttc ggatttcggc ttgtcccgag atgtttatga agaggattcc 4200
tacgtgaaga ggagccaggg tcggattcca gttaaatgga tggcaattga atcccttttt 4260
gatcatatct acaccacgca aagtgatgta tggtcttttg gtgtcctgct gtgggagatc 4320
gtgaccctag ggggaaaccc ctatcctggg attcctcctg agcggctctt caaccttctg 4380
aagaccggcc accggatgga gaggccagac aactgcagcg aggagatgta ccgcctgatg 4440
ctgcaatgct ggaagcagga gccggacaaa aggccggtgt ttgcggacat cagcaaagac 4500
ctggagaaga tgatggttaa gaggagagac tacttggacc ttgcggcgtc cactccatct 4560
gactccctga tttatgacga cggcctctca gaggaggaga caccgctggt ggactgtaat 4620
aatgcccccc tccctcgagc cctcccttcc acatggattg aaaacaaact ctatggcatg 4680
tcagacccga actggcctgg agagagtcct gtaccactca cgagagctga tggcactaac 4740
actgggtttc caagatatcc aaatgatagt gtatatgcta actggatgct ttcaccctca 4800
gcggcaaaat taatggacac gtttgatagt taa 4833
<210> 10
<211> 1610
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 10
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser
130 135 140
Ser Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser
145 150 155 160
Ser Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile
165 170 175
Pro Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser
180 185 190
Pro Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu
195 200 205
Glu Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys
210 215 220
Leu Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu
225 230 235 240
Trp Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu
245 250 255
Lys Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg
260 265 270
Tyr Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr
275 280 285
Leu Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu
290 295 300
Val Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu
305 310 315 320
Ile Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser
325 330 335
Ser Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp
340 345 350
Ala Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His
355 360 365
Val Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val
370 375 380
Val Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu
385 390 395 400
Ser Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala
405 410 415
Glu Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu
420 425 430
Lys Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn
435 440 445
Glu Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu
450 455 460
Leu Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln
465 470 475 480
Lys Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr
485 490 495
Ile Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg
500 505 510
Glu Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro
515 520 525
Pro Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala
530 535 540
His Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln
545 550 555 560
Ala Asn Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe
565 570 575
Leu Arg Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met
580 585 590
Pro Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu
595 600 605
Lys Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly
610 615 620
Ala Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu
625 630 635 640
Ser Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln
645 650 655
Cys Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met
660 665 670
Ser Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys
675 680 685
Asp Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile
690 695 700
Lys Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu
705 710 715 720
Asp Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala
725 730 735
Leu Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln
740 745 750
Gly Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu
755 760 765
Thr Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg
770 775 780
Met Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly
785 790 795 800
Pro Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp
805 810 815
Val Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile
820 825 830
Ala Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu
835 840 845
Leu Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser
850 855 860
Pro Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met
865 870 875 880
Asn Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg
885 890 895
Pro Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg
900 905 910
Ala Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg
915 920 925
Glu Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu
930 935 940
Glu Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu
945 950 955 960
Asp Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr
965 970 975
Arg Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe
980 985 990
Glu Glu Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala
995 1000 1005
Glu Lys Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg
1010 1015 1020
Thr Ile Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr
1025 1030 1035
Leu Val Ser Gly Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro
1040 1045 1050
Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu
1055 1060 1065
Leu Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln
1070 1075 1080
His Glu Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu
1085 1090 1095
Ala Ser Leu Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly
1100 1105 1110
Pro Gly Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser
1115 1120 1125
Gln Leu Leu Glu Thr Leu Asn Gln Leu Ser Thr His Thr His Val
1130 1135 1140
Val Asp Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala
1145 1150 1155
Gln Leu Met Glu Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr
1160 1165 1170
Val Glu Lys Leu Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln
1175 1180 1185
Arg Pro Gly Ala Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser
1190 1195 1200
Ser Ala Phe Leu Arg Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys
1205 1210 1215
Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys
1220 1225 1230
Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr
1235 1240 1245
Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser
1250 1255 1260
Glu Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val
1265 1270 1275
Asn His Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp
1280 1285 1290
Gly Pro Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu
1295 1300 1305
Arg Gly Phe Leu Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu
1310 1315 1320
Gly Ser Gly Gly Ser Arg Asn Ser Ser Ser Leu Asp His Pro Asp
1325 1330 1335
Glu Arg Ala Leu Thr Met Gly Asp Leu Ile Ser Phe Ala Trp Gln
1340 1345 1350
Ile Ser Gln Gly Met Gln Tyr Leu Ala Glu Met Lys Leu Val His
1355 1360 1365
Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Ala Glu Gly Arg Lys
1370 1375 1380
Met Lys Ile Ser Asp Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu
1385 1390 1395
Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg Ile Pro Val Lys Trp
1400 1405 1410
Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr Thr Thr Gln Ser
1415 1420 1425
Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile Val Thr Leu
1430 1435 1440
Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn
1445 1450 1455
Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser
1460 1465 1470
Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro
1475 1480 1485
Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys
1490 1495 1500
Met Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr
1505 1510 1515
Pro Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu
1520 1525 1530
Thr Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu
1535 1540 1545
Pro Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Met Ser Asp Pro
1550 1555 1560
Asn Trp Pro Gly Glu Ser Pro Val Pro Leu Thr Arg Ala Asp Gly
1565 1570 1575
Thr Asn Thr Gly Phe Pro Arg Tyr Pro Asn Asp Ser Val Tyr Ala
1580 1585 1590
Asn Trp Met Leu Ser Pro Ser Ala Ala Lys Leu Met Asp Thr Phe
1595 1600 1605
Asp Ser
1610
<210> 11
<211> 4707
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 11
atggcacaga gcaagaggca cgtgtacagc cggacgccca gcggcagcag gatgagtgcg 60
gaggcaagcg cccggcctct gcgggtgggc tcccgtgtag aggtgattgg aaaaggccac 120
cgaggcactg tggcctatgt tggagccaca ctgtttgcca ctggcaaatg ggtaggcgtg 180
attctggatg aagcaaaggg caaaaatgat ggaactgttc aaggcaggaa gtacttcact 240
tgtgatgaag ggcatggcat ctttgtgcgc cagtcccaga tccaggtatt tgaagatgga 300
gcagatacta cttccccaga gacacctgat tcttctgctt caaaagtcct caaaagagag 360
ggaactgata caactgcaaa gactagcaaa ctgcccacgc gcccagccag tactggggtg 420
gctggggcca gtagctccct gggcccctct ggctcagcgt cagcaggtga gctgagcagc 480
agtgagccca gcaccccggc tcagactccg ctggcagcac ccatcatccc cacgccggtc 540
ctcacctctc ctggagcagt ccccccgctt ccttccccat ccaaggagga ggagggacta 600
agggctcagg tgcgggacct ggaggagaaa ctagagaccc tgagactgaa acgggcagaa 660
gacaaagcaa agctaaaaga gctggagaaa cacaaaatcc agctggagca ggtgcaggaa 720
tggaagagca aaatgcagga gcagcaggcc gacctgcagc ggcgcctcaa ggaggcgaga 780
aaggaagcca aggaggcgct ggaggcaaag gaacgctata tggaggagat ggctgatact 840
gctgatgcca ttgagatggc cactttggac aaggagatgg ctgaagagcg ggctgagtcc 900
ctgcagcagg aggtggaggc actgaaggag cgggtggacg agctcactac tgacttagag 960
atcctcaagg ctgagattga agagaagggc tcagatggcg ctgcatccag ttatcagctc 1020
aagcagcttg aggagcagaa tgcccgcctg aaggatgccc tggtgaggat gcgggatctt 1080
tcttcctcag agaagcagga gcatgtgaag ctccagaagc tcatggaaaa gaagaaccaa 1140
gagctggaag ttgtgaggca acagcgggag cgtctgcagg aggagctaag ccaggcagag 1200
agcaccattg atgagctcaa ggagcaggtg gatgctgctc tgggtgctga ggagatggtg 1260
gagatgctga cagatcggaa cctgaatctg gaagagaaag tgcgcgagtt gagggagact 1320
gtgggagact tggaagcgat gaatgagatg aacgatgagc tgcaggagaa tgcacgtgag 1380
acagaactgg agctgcggga gcagctggac atggcaggcg cgcgggttcg tgaggcccag 1440
aagcgtgtgg aggcagccca ggagacggtt gcagactacc agcagaccat caagaagtac 1500
cgccagctga ccgcccatct acaggatgtg aatcgggaac tgacaaacca gcaggaagca 1560
tctgtggaga ggcaacagca gccacctcca gagacctttg acttcaaaat caagtttgct 1620
gagactaagg cccatgccaa ggcaattgag atggaattga ggcagatgga ggtggcccag 1680
gccaatcgac acatgtccct gctgacagcc ttcatgcctg acagcttcct tcggccaggt 1740
ggggaccatg actgcgttct ggtgctgttg ctcatgcctc gtctcatttg caaggcagag 1800
ctgatccgga agcaggccca ggagaagttt gaactaagtg agaactgttc agagcggcct 1860
gggctgcgag gagctgctgg ggagcaactc agctttgctg ctggactggt gtactcgctg 1920
agcctgctgc aggccacgct acaccgctat gagcatgccc tctctcagtg cagtgtggat 1980
gtgtataaga aagtgggcag cctgtaccct gagatgagtg cccatgagcg ctccttggat 2040
ttcctcattg aactgctgca caaggatcag ctggatgaga ctgtcaatgt ggagcctctc 2100
accaaggcca tcaagtacta tcagcatctg tacagcatcc accttgccga acagcctgag 2160
gactgtacta tgcagctggc tgaccacatt aagttcacgc agagtgctct ggactgcatg 2220
agtgtggagg taggacggct gcgtgccttc ttgcagggtg ggcaggaggc tacagatatt 2280
gccctcctgc tccgggatct ggaaacttca tgcagtgaca tccgccagtt ctgcaagaag 2340
atccgaaggc gaatgccagg gacagatgct cctgggatcc cagctgcact ggcctttgga 2400
ccacaggtat ctgacacgct cctagactgc aggaaacact tgacgtgggt cgtggctgtg 2460
ctgcaggagg tggcagctgc tgctgcccag ctcattgccc cactggcaga gaatgagggg 2520
ctacttgtgg ctgctctgga ggaactggct ttcaaagcaa gcgagcagat ctatgggacc 2580
ccctccagca gcccctatga gtgtctgcgc cagtcatgca acatcctcat cagtaccatg 2640
aacaagctgg ccacagccat gcaggagggg gagtatgatg cagagcggcc ccccagcaag 2700
cctccaccgg ttgaactgcg ggctgctgcc cttcgtgcag agatcacaga tgctgaaggc 2760
ctgggtttga agctcgaaga tcgagagaca gttattaagg agttgaagaa gtcactcaag 2820
attaagggag aggagctaag tgaggccaat gtgcggctga gcctcctgga gaagaagttg 2880
gacagtgctg ccaaggatgc agatgagcgc atcgagaaag tccagactcg gctggaggag 2940
acccaggcac tgctgcgaaa gaaggagaaa gagtttgagg agacaatgga tgcactccag 3000
gctgacatcg accagctgga ggcagagaag gcagaactaa agcagcgtct gaacagccag 3060
tccaaacgca cgattgaggg actccggggc cctcctcctt caggcattgc tactctggtc 3120
tctggcattg ctggtggagc catccctggg caggctccag ggtctgtgcc aggcccaggg 3180
ctggtgaagg actcaccact gctgcttcag cagatctctg ccatgaggct gcacatctcc 3240
cagctccagc atgagaacag catcctcaag ggagcccaga tgaaggcatc cttggcatcc 3300
ctgccccctc tgcatgttgc aaagctatcc catgagggcc ctggcagtga gttaccagct 3360
ggagcgctgt atcgtaagac cagccagctg ctggagacat tgaatcaatt gagcacacac 3420
acgcacgtag tagacatcac tcgcaccagc cctgctgcca agagcccgtc ggcccaactt 3480
atggagcaag tggctcagct taagtccctg agtgacaccg tcgagaagct caaggatgag 3540
gtcctcaagg agacagtatc tcagcgccct ggagccacag tacccactga ctttgccacc 3600
ttcccttcat cagccttcct cagggaggat ccaaagtggg aattccctcg gaagaacttg 3660
gttcttggaa aaactctagg agaaggcgaa tttggaaaag tggtcaaggc aacggccttc 3720
catctgaaag gcagagcagg gtacaccacg gtggccgtga agatgctgaa agagaacgcc 3780
tccccgagtg agcttcgaga cctgctgtca gagttcaacg tcctgaagca ggtcaaccac 3840
ccacatgtca tcaaattgta tggggcctgc agccaggatg gcccgctcct cctcatcgtg 3900
gagtacgcca aatacggctc cctgcggggc ttcctccgcg agagccgcaa agtggggcct 3960
ggctacctgg gcagtggagg cagccgcaac tccagctccc tggaccaccc ggatgagcgg 4020
gccctcacca tgggcgacct catctcattt gcctggcaga tctcacaggg gatgcagtat 4080
ctggccgaga tgaagctcgt tcatcgggac ttggcagcca gaaacatcct ggtagctgag 4140
gggcggaaga tgaagatttc ggatttcggc ttgtcccgag atgtttatga agaggattcc 4200
tacgtgaaga ggagccaggg tcggattcca gttaaatgga tggcaattga atcccttttt 4260
gatcatatct acaccacgca aagtgatgta tggtcttttg gtgtcctgct gtgggagatc 4320
gtgaccctag ggggaaaccc ctatcctggg attcctcctg agcggctctt caaccttctg 4380
aagaccggcc accggatgga gaggccagac aactgcagcg aggagatgta ccgcctgatg 4440
ctgcaatgct ggaagcagga gccggacaaa aggccggtgt ttgcggacat cagcaaagac 4500
ctggagaaga tgatggttaa gaggagagac tacttggacc ttgcggcgtc cactccatct 4560
gactccctga tttatgacga cggcctctca gaggaggaga caccgctggt ggactgtaat 4620
aatgcccccc tccctcgagc cctcccttcc acatggattg aaaacaaact ctatggtaga 4680
atttcccatg catttactag attctag 4707
<210> 12
<211> 1568
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 12
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser
130 135 140
Ser Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser
145 150 155 160
Ser Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile
165 170 175
Pro Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser
180 185 190
Pro Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu
195 200 205
Glu Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys
210 215 220
Leu Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu
225 230 235 240
Trp Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu
245 250 255
Lys Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg
260 265 270
Tyr Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr
275 280 285
Leu Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu
290 295 300
Val Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu
305 310 315 320
Ile Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser
325 330 335
Ser Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp
340 345 350
Ala Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His
355 360 365
Val Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val
370 375 380
Val Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu
385 390 395 400
Ser Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala
405 410 415
Glu Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu
420 425 430
Lys Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn
435 440 445
Glu Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu
450 455 460
Leu Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln
465 470 475 480
Lys Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr
485 490 495
Ile Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg
500 505 510
Glu Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro
515 520 525
Pro Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala
530 535 540
His Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln
545 550 555 560
Ala Asn Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe
565 570 575
Leu Arg Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met
580 585 590
Pro Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu
595 600 605
Lys Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly
610 615 620
Ala Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu
625 630 635 640
Ser Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln
645 650 655
Cys Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met
660 665 670
Ser Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys
675 680 685
Asp Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile
690 695 700
Lys Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu
705 710 715 720
Asp Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala
725 730 735
Leu Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln
740 745 750
Gly Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu
755 760 765
Thr Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg
770 775 780
Met Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly
785 790 795 800
Pro Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp
805 810 815
Val Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile
820 825 830
Ala Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu
835 840 845
Leu Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser
850 855 860
Pro Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met
865 870 875 880
Asn Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg
885 890 895
Pro Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg
900 905 910
Ala Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg
915 920 925
Glu Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu
930 935 940
Glu Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu
945 950 955 960
Asp Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr
965 970 975
Arg Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe
980 985 990
Glu Glu Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala
995 1000 1005
Glu Lys Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg
1010 1015 1020
Thr Ile Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr
1025 1030 1035
Leu Val Ser Gly Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro
1040 1045 1050
Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu
1055 1060 1065
Leu Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln
1070 1075 1080
His Glu Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu
1085 1090 1095
Ala Ser Leu Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly
1100 1105 1110
Pro Gly Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser
1115 1120 1125
Gln Leu Leu Glu Thr Leu Asn Gln Leu Ser Thr His Thr His Val
1130 1135 1140
Val Asp Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala
1145 1150 1155
Gln Leu Met Glu Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr
1160 1165 1170
Val Glu Lys Leu Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln
1175 1180 1185
Arg Pro Gly Ala Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser
1190 1195 1200
Ser Ala Phe Leu Arg Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys
1205 1210 1215
Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys
1220 1225 1230
Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr
1235 1240 1245
Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser
1250 1255 1260
Glu Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val
1265 1270 1275
Asn His Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp
1280 1285 1290
Gly Pro Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu
1295 1300 1305
Arg Gly Phe Leu Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu
1310 1315 1320
Gly Ser Gly Gly Ser Arg Asn Ser Ser Ser Leu Asp His Pro Asp
1325 1330 1335
Glu Arg Ala Leu Thr Met Gly Asp Leu Ile Ser Phe Ala Trp Gln
1340 1345 1350
Ile Ser Gln Gly Met Gln Tyr Leu Ala Glu Met Lys Leu Val His
1355 1360 1365
Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Ala Glu Gly Arg Lys
1370 1375 1380
Met Lys Ile Ser Asp Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu
1385 1390 1395
Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg Ile Pro Val Lys Trp
1400 1405 1410
Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr Thr Thr Gln Ser
1415 1420 1425
Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile Val Thr Leu
1430 1435 1440
Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn
1445 1450 1455
Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser
1460 1465 1470
Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro
1475 1480 1485
Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys
1490 1495 1500
Met Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr
1505 1510 1515
Pro Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu
1520 1525 1530
Thr Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu
1535 1540 1545
Pro Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Arg Ile Ser His
1550 1555 1560
Ala Phe Thr Arg Phe
1565
<210> 13
<211> 4491
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 13
atgatgagac aggcaccgac agcccgaaag accacaactc ggcgacccaa gcccacgcgc 60
ccagccagta ctggggtggc tggggccagt agctccctgg gcccctctgg ctcagcgtca 120
gcaggtgagc tgagcagcag tgagcccagc accccggctc agactccgct ggcagcaccc 180
atcatcccca cgccggtcct cacctctcct ggagcagtcc ccccgcttcc ttccccatcc 240
aaggaggagg agggactaag ggctcaggtg cgggacctgg aggagaaact agagaccctg 300
agactgaaac gggcagaaga caaagcaaag ctaaaagagc tggagaaaca caaaatccag 360
ctggagcagg tgcaggaatg gaagagcaaa atgcaggagc agcaggccga cctgcagcgg 420
cgcctcaagg aggcgagaaa ggaagccaag gaggcgctgg aggcaaagga acgctatatg 480
gaggagatgg ctgatactgc tgatgccatt gagatggcca ctttggacaa ggagatggct 540
gaagagcggg ctgagtccct gcagcaggag gtggaggcac tgaaggagcg ggtggacgag 600
ctcactactg acttagagat cctcaaggct gagattgaag agaagggctc agatggcgct 660
gcatccagtt atcagctcaa gcagcttgag gagcagaatg cccgcctgaa ggatgccctg 720
gtgaggatgc gggatctttc ttcctcagag aagcaggagc atgtgaagct ccagaagctc 780
atggaaaaga agaaccaaga gctggaagtt gtgaggcaac agcgggagcg tctgcaggag 840
gagctaagcc aggcagagag caccattgat gagctcaagg agcaggtgga tgctgctctg 900
ggtgctgagg agatggtgga gatgctgaca gatcggaacc tgaatctgga agagaaagtg 960
cgcgagttga gggagactgt gggagacttg gaagcgatga atgagatgaa cgatgagctg 1020
caggagaatg cacgtgagac agaactggag ctgcgggagc agctggacat ggcaggcgcg 1080
cgggttcgtg aggcccagaa gcgtgtggag gcagcccagg agacggttgc agactaccag 1140
cagaccatca agaagtaccg ccagctgacc gcccatctac aggatgtgaa tcgggaactg 1200
acaaaccagc aggaagcatc tgtggagagg caacagcagc cacctccaga gacctttgac 1260
ttcaaaatca agtttgctga gactaaggcc catgccaagg caattgagat ggaattgagg 1320
cagatggagg tggcccaggc caatcgacac atgtccctgc tgacagcctt catgcctgac 1380
agcttccttc ggccaggtgg ggaccatgac tgcgttctgg tgctgttgct catgcctcgt 1440
ctcatttgca aggcagagct gatccggaag caggcccagg agaagtttga actaagtgag 1500
aactgttcag agcggcctgg gctgcgagga gctgctgggg agcaactcag ctttgctgct 1560
ggactggtgt actcgctgag cctgctgcag gccacgctac accgctatga gcatgccctc 1620
tctcagtgca gtgtggatgt gtataagaaa gtgggcagcc tgtaccctga gatgagtgcc 1680
catgagcgct ccttggattt cctcattgaa ctgctgcaca aggatcagct ggatgagact 1740
gtcaatgtgg agcctctcac caaggccatc aagtactatc agcatctgta cagcatccac 1800
cttgccgaac agcctgagga ctgtactatg cagctggctg accacattaa gttcacgcag 1860
agtgctctgg actgcatgag tgtggaggta ggacggctgc gtgccttctt gcagggtggg 1920
caggaggcta cagatattgc cctcctgctc cgggatctgg aaacttcatg cagtgacatc 1980
cgccagttct gcaagaagat ccgaaggcga atgccaggga cagatgctcc tgggatccca 2040
gctgcactgg cctttggacc acaggtatct gacacgctcc tagactgcag gaaacacttg 2100
acgtgggtcg tggctgtgct gcaggaggtg gcagctgctg ctgcccagct cattgcccca 2160
ctggcagaga atgaggggct acttgtggct gctctggagg aactggcttt caaagcaagc 2220
gagcagatct atgggacccc ctccagcagc ccctatgagt gtctgcgcca gtcatgcaac 2280
atcctcatca gtaccatgaa caagctggcc acagccatgc aggaggggga gtatgatgca 2340
gagcggcccc ccagcaagcc tccaccggtt gaactgcggg ctgctgccct tcgtgcagag 2400
atcacagatg ctgaaggcct gggtttgaag ctcgaagatc gagagacagt tattaaggag 2460
ttgaagaagt cactcaagat taagggagag gagctaagtg aggccaatgt gcggctgagc 2520
ctcctggaga agaagttgga cagtgctgcc aaggatgcag atgagcgcat cgagaaagtc 2580
cagactcggc tggaggagac ccaggcactg ctgcgaaaga aggagaaaga gtttgaggag 2640
acaatggatg cactccaggc tgacatcgac cagctggagg cagagaaggc agaactaaag 2700
cagcgtctga acagccagtc caaacgcacg attgagggac tccggggccc tcctccttca 2760
ggcattgcta ctctggtctc tggcattgct ggtggagcca tccctgggca ggctccaggg 2820
tctgtgccag gcccagggct ggtgaaggac tcaccactgc tgcttcagca gatctctgcc 2880
atgaggctgc acatctccca gctccagcat gagaacagca tcctcaaggg agcccagatg 2940
aaggcatcct tggcatccct gccccctctg catgttgcaa agctatccca tgagggccct 3000
ggcagtgagt taccagctgg agcgctgtat cgtaagacca gccagctgct ggagacattg 3060
aatcaattga gcacacacac gcacgtagta gacatcactc gcaccagccc tgctgccaag 3120
agcccgtcgg cccaacttat ggagcaagtg gctcagctta agtccctgag tgacaccgtc 3180
gagaagctca aggatgaggt cctcaaggag acagtatctc agcgccctgg agccacagta 3240
cccactgact ttgccacctt cccttcatca gccttcctca gggaggatcc aaagtgggaa 3300
ttccctcgga agaacttggt tcttggaaaa actctaggag aaggcgaatt tggaaaagtg 3360
gtcaaggcaa cggccttcca tctgaaaggc agagcagggt acaccacggt ggccgtgaag 3420
atgctgaaag agaacgcctc cccgagtgag cttcgagacc tgctgtcaga gttcaacgtc 3480
ctgaagcagg tcaaccaccc acatgtcatc aaattgtatg gggcctgcag ccaggatggc 3540
ccgctcctcc tcatcgtgga gtacgccaaa tacggctccc tgcggggctt cctccgcgag 3600
agccgcaaag tggggcctgg ctacctgggc agtggaggca gccgcaactc cagctccctg 3660
gaccacccgg atgagcgggc cctcaccatg ggcgacctca tctcatttgc ctggcagatc 3720
tcacagggga tgcagtatct ggccgagatg aagctcgttc atcgggactt ggcagccaga 3780
aacatcctgg tagctgaggg gcggaagatg aagatttcgg atttcggctt gtcccgagat 3840
gtttatgaag aggattccta cgtgaagagg agccagggtc ggattccagt taaatggatg 3900
gcaattgaat ccctttttga tcatatctac accacgcaaa gtgatgtatg gtcttttggt 3960
gtcctgctgt gggagatcgt gaccctaggg ggaaacccct atcctgggat tcctcctgag 4020
cggctcttca accttctgaa gaccggccac cggatggaga ggccagacaa ctgcagcgag 4080
gagatgtacc gcctgatgct gcaatgctgg aagcaggagc cggacaaaag gccggtgttt 4140
gcggacatca gcaaagacct ggagaagatg atggttaaga ggagagacta cttggacctt 4200
gcggcgtcca ctccatctga ctccctgatt tatgacgacg gcctctcaga ggaggagaca 4260
ccgctggtgg actgtaataa tgcccccctc cctcgagccc tcccttccac atggattgaa 4320
aacaaactct atggcatgtc agacccgaac tggcctggag agagtcctgt accactcacg 4380
agagctgatg gcactaacac tgggtttcca agatatccaa atgatagtgt atatgctaac 4440
tggatgcttt caccctcagc ggcaaaatta atggacacgt ttgatagtta a 4491
<210> 14
<211> 1496
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 14
Met Met Arg Gln Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro
1 5 10 15
Lys Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser
20 25 30
Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu
35 40 45
Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr
50 55 60
Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser
65 70 75 80
Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys
85 90 95
Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys
100 105 110
Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys
115 120 125
Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu
130 135 140
Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met
145 150 155 160
Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp
165 170 175
Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu
180 185 190
Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu
195 200 205
Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr
210 215 220
Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu
225 230 235 240
Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys
245 250 255
Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg
260 265 270
Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr
275 280 285
Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu
290 295 300
Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val
305 310 315 320
Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met
325 330 335
Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg
340 345 350
Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg
355 360 365
Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys
370 375 380
Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu
385 390 395 400
Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro
405 410 415
Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala
420 425 430
Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn
435 440 445
Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg
450 455 460
Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg
465 470 475 480
Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe
485 490 495
Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala
500 505 510
Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu
515 520 525
Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser
530 535 540
Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala
545 550 555 560
His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln
565 570 575
Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr
580 585 590
Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys
595 600 605
Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp
610 615 620
Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly
625 630 635 640
Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser
645 650 655
Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro
660 665 670
Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln
675 680 685
Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val
690 695 700
Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro
705 710 715 720
Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala
725 730 735
Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr
740 745 750
Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys
755 760 765
Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro
770 775 780
Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu
785 790 795 800
Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr
805 810 815
Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu
820 825 830
Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser
835 840 845
Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu
850 855 860
Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu
865 870 875 880
Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
885 890 895
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile Glu
900 905 910
Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val Ser Gly
915 920 925
Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser Val Pro Gly
930 935 940
Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln Gln Ile Ser Ala
945 950 955 960
Met Arg Leu His Ile Ser Gln Leu Gln His Glu Asn Ser Ile Leu Lys
965 970 975
Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu Pro Pro Leu His Val
980 985 990
Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu Leu Pro Ala Gly Ala
995 1000 1005
Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu Thr Leu Asn Gln Leu
1010 1015 1020
Ser Thr His Thr His Val Val Asp Ile Thr Arg Thr Ser Pro Ala
1025 1030 1035
Ala Lys Ser Pro Ser Ala Gln Leu Met Glu Gln Val Ala Gln Leu
1040 1045 1050
Lys Ser Leu Ser Asp Thr Val Glu Lys Leu Lys Asp Glu Val Leu
1055 1060 1065
Lys Glu Thr Val Ser Gln Arg Pro Gly Ala Thr Val Pro Thr Asp
1070 1075 1080
Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu Arg Glu Asp Pro Lys
1085 1090 1095
Trp Glu Phe Pro Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly
1100 1105 1110
Glu Gly Glu Phe Gly Lys Val Val Lys Ala Thr Ala Phe His Leu
1115 1120 1125
Lys Gly Arg Ala Gly Tyr Thr Thr Val Ala Val Lys Met Leu Lys
1130 1135 1140
Glu Asn Ala Ser Pro Ser Glu Leu Arg Asp Leu Leu Ser Glu Phe
1145 1150 1155
Asn Val Leu Lys Gln Val Asn His Pro His Val Ile Lys Leu Tyr
1160 1165 1170
Gly Ala Cys Ser Gln Asp Gly Pro Leu Leu Leu Ile Val Glu Tyr
1175 1180 1185
Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu Arg Glu Ser Arg Lys
1190 1195 1200
Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser Arg Asn Ser Ser
1205 1210 1215
Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr Met Gly Asp Leu
1220 1225 1230
Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr Leu Ala
1235 1240 1245
Glu Met Lys Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile Leu
1250 1255 1260
Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser
1265 1270 1275
Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly
1280 1285 1290
Arg Ile Pro Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His
1295 1300 1305
Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu
1310 1315 1320
Trp Glu Ile Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro
1325 1330 1335
Pro Glu Arg Leu Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu
1340 1345 1350
Arg Pro Asp Asn Cys Ser Glu Glu Met Tyr Arg Leu Met Leu Gln
1355 1360 1365
Cys Trp Lys Gln Glu Pro Asp Lys Arg Pro Val Phe Ala Asp Ile
1370 1375 1380
Ser Lys Asp Leu Glu Lys Met Met Val Lys Arg Arg Asp Tyr Leu
1385 1390 1395
Asp Leu Ala Ala Ser Thr Pro Ser Asp Ser Leu Ile Tyr Asp Asp
1400 1405 1410
Gly Leu Ser Glu Glu Glu Thr Pro Leu Val Asp Cys Asn Asn Ala
1415 1420 1425
Pro Leu Pro Arg Ala Leu Pro Ser Thr Trp Ile Glu Asn Lys Leu
1430 1435 1440
Tyr Gly Met Ser Asp Pro Asn Trp Pro Gly Glu Ser Pro Val Pro
1445 1450 1455
Leu Thr Arg Ala Asp Gly Thr Asn Thr Gly Phe Pro Arg Tyr Pro
1460 1465 1470
Asn Asp Ser Val Tyr Ala Asn Trp Met Leu Ser Pro Ser Ala Ala
1475 1480 1485
Lys Leu Met Asp Thr Phe Asp Ser
1490 1495
<210> 15
<211> 4365
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 15
atgatgagac aggcaccgac agcccgaaag accacaactc ggcgacccaa gcccacgcgc 60
ccagccagta ctggggtggc tggggccagt agctccctgg gcccctctgg ctcagcgtca 120
gcaggtgagc tgagcagcag tgagcccagc accccggctc agactccgct ggcagcaccc 180
atcatcccca cgccggtcct cacctctcct ggagcagtcc ccccgcttcc ttccccatcc 240
aaggaggagg agggactaag ggctcaggtg cgggacctgg aggagaaact agagaccctg 300
agactgaaac gggcagaaga caaagcaaag ctaaaagagc tggagaaaca caaaatccag 360
ctggagcagg tgcaggaatg gaagagcaaa atgcaggagc agcaggccga cctgcagcgg 420
cgcctcaagg aggcgagaaa ggaagccaag gaggcgctgg aggcaaagga acgctatatg 480
gaggagatgg ctgatactgc tgatgccatt gagatggcca ctttggacaa ggagatggct 540
gaagagcggg ctgagtccct gcagcaggag gtggaggcac tgaaggagcg ggtggacgag 600
ctcactactg acttagagat cctcaaggct gagattgaag agaagggctc agatggcgct 660
gcatccagtt atcagctcaa gcagcttgag gagcagaatg cccgcctgaa ggatgccctg 720
gtgaggatgc gggatctttc ttcctcagag aagcaggagc atgtgaagct ccagaagctc 780
atggaaaaga agaaccaaga gctggaagtt gtgaggcaac agcgggagcg tctgcaggag 840
gagctaagcc aggcagagag caccattgat gagctcaagg agcaggtgga tgctgctctg 900
ggtgctgagg agatggtgga gatgctgaca gatcggaacc tgaatctgga agagaaagtg 960
cgcgagttga gggagactgt gggagacttg gaagcgatga atgagatgaa cgatgagctg 1020
caggagaatg cacgtgagac agaactggag ctgcgggagc agctggacat ggcaggcgcg 1080
cgggttcgtg aggcccagaa gcgtgtggag gcagcccagg agacggttgc agactaccag 1140
cagaccatca agaagtaccg ccagctgacc gcccatctac aggatgtgaa tcgggaactg 1200
acaaaccagc aggaagcatc tgtggagagg caacagcagc cacctccaga gacctttgac 1260
ttcaaaatca agtttgctga gactaaggcc catgccaagg caattgagat ggaattgagg 1320
cagatggagg tggcccaggc caatcgacac atgtccctgc tgacagcctt catgcctgac 1380
agcttccttc ggccaggtgg ggaccatgac tgcgttctgg tgctgttgct catgcctcgt 1440
ctcatttgca aggcagagct gatccggaag caggcccagg agaagtttga actaagtgag 1500
aactgttcag agcggcctgg gctgcgagga gctgctgggg agcaactcag ctttgctgct 1560
ggactggtgt actcgctgag cctgctgcag gccacgctac accgctatga gcatgccctc 1620
tctcagtgca gtgtggatgt gtataagaaa gtgggcagcc tgtaccctga gatgagtgcc 1680
catgagcgct ccttggattt cctcattgaa ctgctgcaca aggatcagct ggatgagact 1740
gtcaatgtgg agcctctcac caaggccatc aagtactatc agcatctgta cagcatccac 1800
cttgccgaac agcctgagga ctgtactatg cagctggctg accacattaa gttcacgcag 1860
agtgctctgg actgcatgag tgtggaggta ggacggctgc gtgccttctt gcagggtggg 1920
caggaggcta cagatattgc cctcctgctc cgggatctgg aaacttcatg cagtgacatc 1980
cgccagttct gcaagaagat ccgaaggcga atgccaggga cagatgctcc tgggatccca 2040
gctgcactgg cctttggacc acaggtatct gacacgctcc tagactgcag gaaacacttg 2100
acgtgggtcg tggctgtgct gcaggaggtg gcagctgctg ctgcccagct cattgcccca 2160
ctggcagaga atgaggggct acttgtggct gctctggagg aactggcttt caaagcaagc 2220
gagcagatct atgggacccc ctccagcagc ccctatgagt gtctgcgcca gtcatgcaac 2280
atcctcatca gtaccatgaa caagctggcc acagccatgc aggaggggga gtatgatgca 2340
gagcggcccc ccagcaagcc tccaccggtt gaactgcggg ctgctgccct tcgtgcagag 2400
atcacagatg ctgaaggcct gggtttgaag ctcgaagatc gagagacagt tattaaggag 2460
ttgaagaagt cactcaagat taagggagag gagctaagtg aggccaatgt gcggctgagc 2520
ctcctggaga agaagttgga cagtgctgcc aaggatgcag atgagcgcat cgagaaagtc 2580
cagactcggc tggaggagac ccaggcactg ctgcgaaaga aggagaaaga gtttgaggag 2640
acaatggatg cactccaggc tgacatcgac cagctggagg cagagaaggc agaactaaag 2700
cagcgtctga acagccagtc caaacgcacg attgagggac tccggggccc tcctccttca 2760
ggcattgcta ctctggtctc tggcattgct ggtggagcca tccctgggca ggctccaggg 2820
tctgtgccag gcccagggct ggtgaaggac tcaccactgc tgcttcagca gatctctgcc 2880
atgaggctgc acatctccca gctccagcat gagaacagca tcctcaaggg agcccagatg 2940
aaggcatcct tggcatccct gccccctctg catgttgcaa agctatccca tgagggccct 3000
ggcagtgagt taccagctgg agcgctgtat cgtaagacca gccagctgct ggagacattg 3060
aatcaattga gcacacacac gcacgtagta gacatcactc gcaccagccc tgctgccaag 3120
agcccgtcgg cccaacttat ggagcaagtg gctcagctta agtccctgag tgacaccgtc 3180
gagaagctca aggatgaggt cctcaaggag acagtatctc agcgccctgg agccacagta 3240
cccactgact ttgccacctt cccttcatca gccttcctca gggaggatcc aaagtgggaa 3300
ttccctcgga agaacttggt tcttggaaaa actctaggag aaggcgaatt tggaaaagtg 3360
gtcaaggcaa cggccttcca tctgaaaggc agagcagggt acaccacggt ggccgtgaag 3420
atgctgaaag agaacgcctc cccgagtgag cttcgagacc tgctgtcaga gttcaacgtc 3480
ctgaagcagg tcaaccaccc acatgtcatc aaattgtatg gggcctgcag ccaggatggc 3540
ccgctcctcc tcatcgtgga gtacgccaaa tacggctccc tgcggggctt cctccgcgag 3600
agccgcaaag tggggcctgg ctacctgggc agtggaggca gccgcaactc cagctccctg 3660
gaccacccgg atgagcgggc cctcaccatg ggcgacctca tctcatttgc ctggcagatc 3720
tcacagggga tgcagtatct ggccgagatg aagctcgttc atcgggactt ggcagccaga 3780
aacatcctgg tagctgaggg gcggaagatg aagatttcgg atttcggctt gtcccgagat 3840
gtttatgaag aggattccta cgtgaagagg agccagggtc ggattccagt taaatggatg 3900
gcaattgaat ccctttttga tcatatctac accacgcaaa gtgatgtatg gtcttttggt 3960
gtcctgctgt gggagatcgt gaccctaggg ggaaacccct atcctgggat tcctcctgag 4020
cggctcttca accttctgaa gaccggccac cggatggaga ggccagacaa ctgcagcgag 4080
gagatgtacc gcctgatgct gcaatgctgg aagcaggagc cggacaaaag gccggtgttt 4140
gcggacatca gcaaagacct ggagaagatg atggttaaga ggagagacta cttggacctt 4200
gcggcgtcca ctccatctga ctccctgatt tatgacgacg gcctctcaga ggaggagaca 4260
ccgctggtgg actgtaataa tgcccccctc cctcgagccc tcccttccac atggattgaa 4320
aacaaactct atggtagaat ttcccatgca tttactagat tctag 4365
<210> 16
<211> 1454
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 16
Met Met Arg Gln Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro
1 5 10 15
Lys Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser
20 25 30
Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu
35 40 45
Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr
50 55 60
Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser
65 70 75 80
Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys
85 90 95
Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys
100 105 110
Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys
115 120 125
Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu
130 135 140
Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met
145 150 155 160
Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp
165 170 175
Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu
180 185 190
Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu
195 200 205
Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr
210 215 220
Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu
225 230 235 240
Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys
245 250 255
Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg
260 265 270
Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr
275 280 285
Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu
290 295 300
Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val
305 310 315 320
Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met
325 330 335
Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg
340 345 350
Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg
355 360 365
Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys
370 375 380
Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu
385 390 395 400
Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro
405 410 415
Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala
420 425 430
Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn
435 440 445
Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg
450 455 460
Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg
465 470 475 480
Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe
485 490 495
Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala
500 505 510
Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu
515 520 525
Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser
530 535 540
Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala
545 550 555 560
His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln
565 570 575
Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr
580 585 590
Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys
595 600 605
Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp
610 615 620
Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly
625 630 635 640
Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser
645 650 655
Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro
660 665 670
Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln
675 680 685
Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val
690 695 700
Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro
705 710 715 720
Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala
725 730 735
Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr
740 745 750
Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys
755 760 765
Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro
770 775 780
Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu
785 790 795 800
Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr
805 810 815
Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu
820 825 830
Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser
835 840 845
Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu
850 855 860
Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu
865 870 875 880
Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
885 890 895
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile Glu
900 905 910
Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val Ser Gly
915 920 925
Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser Val Pro Gly
930 935 940
Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln Gln Ile Ser Ala
945 950 955 960
Met Arg Leu His Ile Ser Gln Leu Gln His Glu Asn Ser Ile Leu Lys
965 970 975
Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu Pro Pro Leu His Val
980 985 990
Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu Leu Pro Ala Gly Ala
995 1000 1005
Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu Thr Leu Asn Gln Leu
1010 1015 1020
Ser Thr His Thr His Val Val Asp Ile Thr Arg Thr Ser Pro Ala
1025 1030 1035
Ala Lys Ser Pro Ser Ala Gln Leu Met Glu Gln Val Ala Gln Leu
1040 1045 1050
Lys Ser Leu Ser Asp Thr Val Glu Lys Leu Lys Asp Glu Val Leu
1055 1060 1065
Lys Glu Thr Val Ser Gln Arg Pro Gly Ala Thr Val Pro Thr Asp
1070 1075 1080
Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu Arg Glu Asp Pro Lys
1085 1090 1095
Trp Glu Phe Pro Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly
1100 1105 1110
Glu Gly Glu Phe Gly Lys Val Val Lys Ala Thr Ala Phe His Leu
1115 1120 1125
Lys Gly Arg Ala Gly Tyr Thr Thr Val Ala Val Lys Met Leu Lys
1130 1135 1140
Glu Asn Ala Ser Pro Ser Glu Leu Arg Asp Leu Leu Ser Glu Phe
1145 1150 1155
Asn Val Leu Lys Gln Val Asn His Pro His Val Ile Lys Leu Tyr
1160 1165 1170
Gly Ala Cys Ser Gln Asp Gly Pro Leu Leu Leu Ile Val Glu Tyr
1175 1180 1185
Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu Arg Glu Ser Arg Lys
1190 1195 1200
Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser Arg Asn Ser Ser
1205 1210 1215
Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr Met Gly Asp Leu
1220 1225 1230
Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr Leu Ala
1235 1240 1245
Glu Met Lys Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile Leu
1250 1255 1260
Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser
1265 1270 1275
Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly
1280 1285 1290
Arg Ile Pro Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His
1295 1300 1305
Ile Tyr Thr Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu
1310 1315 1320
Trp Glu Ile Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro
1325 1330 1335
Pro Glu Arg Leu Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu
1340 1345 1350
Arg Pro Asp Asn Cys Ser Glu Glu Met Tyr Arg Leu Met Leu Gln
1355 1360 1365
Cys Trp Lys Gln Glu Pro Asp Lys Arg Pro Val Phe Ala Asp Ile
1370 1375 1380
Ser Lys Asp Leu Glu Lys Met Met Val Lys Arg Arg Asp Tyr Leu
1385 1390 1395
Asp Leu Ala Ala Ser Thr Pro Ser Asp Ser Leu Ile Tyr Asp Asp
1400 1405 1410
Gly Leu Ser Glu Glu Glu Thr Pro Leu Val Asp Cys Asn Asn Ala
1415 1420 1425
Pro Leu Pro Arg Ala Leu Pro Ser Thr Trp Ile Glu Asn Lys Leu
1430 1435 1440
Tyr Gly Arg Ile Ser His Ala Phe Thr Arg Phe
1445 1450
<210> 17
<211> 4782
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 17
atgagtgcgg aggcaagcgc ccggcctctg cgggtgggct cccgtgtaga ggtgattgga 60
aaaggccacc gaggcactgt ggcctatgtt ggagccacac tgtttgccac tggcaaatgg 120
gtaggcgtga ttctggatga agcaaagggc aaaaatgatg gaactgttca aggcaggaag 180
tacttcactt gtgatgaagg gcatggcatc tttgtgcgcc agtcccagat ccaggtattt 240
gaagatggag cagatactac ttccccagag acacctgatt cttctgcttc aaaagtcctc 300
aaaagagagg gaactgatac aactgcaaag actagcaaac tgcccacgcg cccagccagt 360
actggggtgg ctggggccag tagctccctg ggcccctctg gctcagcgtc agcaggtgag 420
ctgagcagca gtgagcccag caccccggct cagactccgc tggcagcacc catcatcccc 480
acgccggtcc tcacctctcc tggagcagtc cccccgcttc cttccccatc caaggaggag 540
gagggactaa gggctcaggt gcgggacctg gaggagaaac tagagaccct gagactgaaa 600
cgggcagaag acaaagcaaa gctaaaagag ctggagaaac acaaaatcca gctggagcag 660
gtgcaggaat ggaagagcaa aatgcaggag cagcaggccg acctgcagcg gcgcctcaag 720
gaggcgagaa aggaagccaa ggaggcgctg gaggcaaagg aacgctatat ggaggagatg 780
gctgatactg ctgatgccat tgagatggcc actttggaca aggagatggc tgaagagcgg 840
gctgagtccc tgcagcagga ggtggaggca ctgaaggagc gggtggacga gctcactact 900
gacttagaga tcctcaaggc tgagattgaa gagaagggct cagatggcgc tgcatccagt 960
tatcagctca agcagcttga ggagcagaat gcccgcctga aggatgccct ggtgaggatg 1020
cgggatcttt cttcctcaga gaagcaggag catgtgaagc tccagaagct catggaaaag 1080
aagaaccaag agctggaagt tgtgaggcaa cagcgggagc gtctgcagga ggagctaagc 1140
caggcagaga gcaccattga tgagctcaag gagcaggtgg atgctgctct gggtgctgag 1200
gagatggtgg agatgctgac agatcggaac ctgaatctgg aagagaaagt gcgcgagttg 1260
agggagactg tgggagactt ggaagcgatg aatgagatga acgatgagct gcaggagaat 1320
gcacgtgaga cagaactgga gctgcgggag cagctggaca tggcaggcgc gcgggttcgt 1380
gaggcccaga agcgtgtgga ggcagcccag gagacggttg cagactacca gcagaccatc 1440
aagaagtacc gccagctgac cgcccatcta caggatgtga atcgggaact gacaaaccag 1500
caggaagcat ctgtggagag gcaacagcag ccacctccag agacctttga cttcaaaatc 1560
aagtttgctg agactaaggc ccatgccaag gcaattgaga tggaattgag gcagatggag 1620
gtggcccagg ccaatcgaca catgtccctg ctgacagcct tcatgcctga cagcttcctt 1680
cggccaggtg gggaccatga ctgcgttctg gtgctgttgc tcatgcctcg tctcatttgc 1740
aaggcagagc tgatccggaa gcaggcccag gagaagtttg aactaagtga gaactgttca 1800
gagcggcctg ggctgcgagg agctgctggg gagcaactca gctttgctgc tggactggtg 1860
tactcgctga gcctgctgca ggccacgcta caccgctatg agcatgccct ctctcagtgc 1920
agtgtggatg tgtataagaa agtgggcagc ctgtaccctg agatgagtgc ccatgagcgc 1980
tccttggatt tcctcattga actgctgcac aaggatcagc tggatgagac tgtcaatgtg 2040
gagcctctca ccaaggccat caagtactat cagcatctgt acagcatcca ccttgccgaa 2100
cagcctgagg actgtactat gcagctggct gaccacatta agttcacgca gagtgctctg 2160
gactgcatga gtgtggaggt aggacggctg cgtgccttct tgcagggtgg gcaggaggct 2220
acagatattg ccctcctgct ccgggatctg gaaacttcat gcagtgacat ccgccagttc 2280
tgcaagaaga tccgaaggcg aatgccaggg acagatgctc ctgggatccc agctgcactg 2340
gcctttggac cacaggtatc tgacacgctc ctagactgca ggaaacactt gacgtgggtc 2400
gtggctgtgc tgcaggaggt ggcagctgct gctgcccagc tcattgcccc actggcagag 2460
aatgaggggc tacttgtggc tgctctggag gaactggctt tcaaagcaag cgagcagatc 2520
tatgggaccc cctccagcag cccctatgag tgtctgcgcc agtcatgcaa catcctcatc 2580
agtaccatga acaagctggc cacagccatg caggaggggg agtatgatgc agagcggccc 2640
cccagcaagc ctccaccggt tgaactgcgg gctgctgccc ttcgtgcaga gatcacagat 2700
gctgaaggcc tgggtttgaa gctcgaagat cgagagacag ttattaagga gttgaagaag 2760
tcactcaaga ttaagggaga ggagctaagt gaggccaatg tgcggctgag cctcctggag 2820
aagaagttgg acagtgctgc caaggatgca gatgagcgca tcgagaaagt ccagactcgg 2880
ctggaggaga cccaggcact gctgcgaaag aaggagaaag agtttgagga gacaatggat 2940
gcactccagg ctgacatcga ccagctggag gcagagaagg cagaactaaa gcagcgtctg 3000
aacagccagt ccaaacgcac gattgaggga ctccggggcc ctcctccttc aggcattgct 3060
actctggtct ctggcattgc tggtggagcc atccctgggc aggctccagg gtctgtgcca 3120
ggcccagggc tggtgaagga ctcaccactg ctgcttcagc agatctctgc catgaggctg 3180
cacatctccc agctccagca tgagaacagc atcctcaagg gagcccagat gaaggcatcc 3240
ttggcatccc tgccccctct gcatgttgca aagctatccc atgagggccc tggcagtgag 3300
ttaccagctg gagcgctgta tcgtaagacc agccagctgc tggagacatt gaatcaattg 3360
agcacacaca cgcacgtagt agacatcact cgcaccagcc ctgctgccaa gagcccgtcg 3420
gcccaactta tggagcaagt ggctcagctt aagtccctga gtgacaccgt cgagaagctc 3480
aaggatgagg tcctcaagga gacagtatct cagcgccctg gagccacagt acccactgac 3540
tttgccacct tcccttcatc agccttcctc agggaggatc caaagtggga attccctcgg 3600
aagaacttgg ttcttggaaa aactctagga gaaggcgaat ttggaaaagt ggtcaaggca 3660
acggccttcc atctgaaagg cagagcaggg tacaccacgg tggccgtgaa gatgctgaaa 3720
gagaacgcct ccccgagtga gcttcgagac ctgctgtcag agttcaacgt cctgaagcag 3780
gtcaaccacc cacatgtcat caaattgtat ggggcctgca gccaggatgg cccgctcctc 3840
ctcatcgtgg agtacgccaa atacggctcc ctgcggggct tcctccgcga gagccgcaaa 3900
gtggggcctg gctacctggg cagtggaggc agccgcaact ccagctccct ggaccacccg 3960
gatgagcggg ccctcaccat gggcgacctc atctcatttg cctggcagat ctcacagggg 4020
atgcagtatc tggccgagat gaagctcgtt catcgggact tggcagccag aaacatcctg 4080
gtagctgagg ggcggaagat gaagatttcg gatttcggct tgtcccgaga tgtttatgaa 4140
gaggattcct acgtgaagag gagccagggt cggattccag ttaaatggat ggcaattgaa 4200
tccctttttg atcatatcta caccacgcaa agtgatgtat ggtcttttgg tgtcctgctg 4260
tgggagatcg tgaccctagg gggaaacccc tatcctggga ttcctcctga gcggctcttc 4320
aaccttctga agaccggcca ccggatggag aggccagaca actgcagcga ggagatgtac 4380
cgcctgatgc tgcaatgctg gaagcaggag ccggacaaaa ggccggtgtt tgcggacatc 4440
agcaaagacc tggagaagat gatggttaag aggagagact acttggacct tgcggcgtcc 4500
actccatctg actccctgat ttatgacgac ggcctctcag aggaggagac accgctggtg 4560
gactgtaata atgcccccct ccctcgagcc ctcccttcca catggattga aaacaaactc 4620
tatggcatgt cagacccgaa ctggcctgga gagagtcctg taccactcac gagagctgat 4680
ggcactaaca ctgggtttcc aagatatcca aatgatagtg tatatgctaa ctggatgctt 4740
tcaccctcag cggcaaaatt aatggacacg tttgatagtt aa 4782
<210> 18
<211> 1593
<212> PRT
<213> 人工序列e
<220>
<223> 融合肽
<400> 18
Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg Val
1 5 10 15
Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly Ala
20 25 30
Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu Ala
35 40 45
Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr Cys
50 55 60
Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val Phe
65 70 75 80
Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser Ala
85 90 95
Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr Ser
100 105 110
Lys Leu Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser
115 120 125
Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser
130 135 140
Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro
145 150 155 160
Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro
165 170 175
Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu
180 185 190
Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu
195 200 205
Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp
210 215 220
Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys
225 230 235 240
Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr
245 250 255
Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu
260 265 270
Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val
275 280 285
Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile
290 295 300
Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser
305 310 315 320
Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala
325 330 335
Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val
340 345 350
Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val
355 360 365
Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser
370 375 380
Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu
385 390 395 400
Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys
405 410 415
Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu
420 425 430
Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu
435 440 445
Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys
450 455 460
Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile
465 470 475 480
Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu
485 490 495
Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro
500 505 510
Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His
515 520 525
Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala
530 535 540
Asn Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu
545 550 555 560
Arg Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro
565 570 575
Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys
580 585 590
Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala
595 600 605
Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser
610 615 620
Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys
625 630 635 640
Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser
645 650 655
Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp
660 665 670
Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys
675 680 685
Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp
690 695 700
Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu
705 710 715 720
Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly
725 730 735
Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr
740 745 750
Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met
755 760 765
Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro
770 775 780
Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val
785 790 795 800
Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala
805 810 815
Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu
820 825 830
Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro
835 840 845
Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn
850 855 860
Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro
865 870 875 880
Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala
885 890 895
Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu
900 905 910
Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu
915 920 925
Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp
930 935 940
Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg
945 950 955 960
Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu
965 970 975
Glu Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu
980 985 990
Lys Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile
995 1000 1005
Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val
1010 1015 1020
Ser Gly Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser
1025 1030 1035
Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln
1040 1045 1050
Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu
1055 1060 1065
Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser
1070 1075 1080
Leu Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly
1085 1090 1095
Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu
1100 1105 1110
Leu Glu Thr Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp
1115 1120 1125
Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu
1130 1135 1140
Met Glu Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu
1145 1150 1155
Lys Leu Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro
1160 1165 1170
Gly Ala Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala
1175 1180 1185
Phe Leu Arg Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn Leu
1190 1195 1200
Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val
1205 1210 1215
Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr
1220 1225 1230
Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu
1235 1240 1245
Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His
1250 1255 1260
Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro
1265 1270 1275
Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly
1280 1285 1290
Phe Leu Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser
1295 1300 1305
Gly Gly Ser Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg
1310 1315 1320
Ala Leu Thr Met Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser
1325 1330 1335
Gln Gly Met Gln Tyr Leu Ala Glu Met Lys Leu Val His Arg Asp
1340 1345 1350
Leu Ala Ala Arg Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys
1355 1360 1365
Ile Ser Asp Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser
1370 1375 1380
Tyr Val Lys Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala
1385 1390 1395
Ile Glu Ser Leu Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val
1400 1405 1410
Trp Ser Phe Gly Val Leu Leu Trp Glu Ile Val Thr Leu Gly Gly
1415 1420 1425
Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu Leu
1430 1435 1440
Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu
1445 1450 1455
Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys
1460 1465 1470
Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met
1475 1480 1485
Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro Ser
1490 1495 1500
Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr Pro
1505 1510 1515
Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro Ser
1520 1525 1530
Thr Trp Ile Glu Asn Lys Leu Tyr Gly Met Ser Asp Pro Asn Trp
1535 1540 1545
Pro Gly Glu Ser Pro Val Pro Leu Thr Arg Ala Asp Gly Thr Asn
1550 1555 1560
Thr Gly Phe Pro Arg Tyr Pro Asn Asp Ser Val Tyr Ala Asn Trp
1565 1570 1575
Met Leu Ser Pro Ser Ala Ala Lys Leu Met Asp Thr Phe Asp Ser
1580 1585 1590
<210> 19
<211> 4656
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 19
atgagtgcgg aggcaagcgc ccggcctctg cgggtgggct cccgtgtaga ggtgattgga 60
aaaggccacc gaggcactgt ggcctatgtt ggagccacac tgtttgccac tggcaaatgg 120
gtaggcgtga ttctggatga agcaaagggc aaaaatgatg gaactgttca aggcaggaag 180
tacttcactt gtgatgaagg gcatggcatc tttgtgcgcc agtcccagat ccaggtattt 240
gaagatggag cagatactac ttccccagag acacctgatt cttctgcttc aaaagtcctc 300
aaaagagagg gaactgatac aactgcaaag actagcaaac tgcccacgcg cccagccagt 360
actggggtgg ctggggccag tagctccctg ggcccctctg gctcagcgtc agcaggtgag 420
ctgagcagca gtgagcccag caccccggct cagactccgc tggcagcacc catcatcccc 480
acgccggtcc tcacctctcc tggagcagtc cccccgcttc cttccccatc caaggaggag 540
gagggactaa gggctcaggt gcgggacctg gaggagaaac tagagaccct gagactgaaa 600
cgggcagaag acaaagcaaa gctaaaagag ctggagaaac acaaaatcca gctggagcag 660
gtgcaggaat ggaagagcaa aatgcaggag cagcaggccg acctgcagcg gcgcctcaag 720
gaggcgagaa aggaagccaa ggaggcgctg gaggcaaagg aacgctatat ggaggagatg 780
gctgatactg ctgatgccat tgagatggcc actttggaca aggagatggc tgaagagcgg 840
gctgagtccc tgcagcagga ggtggaggca ctgaaggagc gggtggacga gctcactact 900
gacttagaga tcctcaaggc tgagattgaa gagaagggct cagatggcgc tgcatccagt 960
tatcagctca agcagcttga ggagcagaat gcccgcctga aggatgccct ggtgaggatg 1020
cgggatcttt cttcctcaga gaagcaggag catgtgaagc tccagaagct catggaaaag 1080
aagaaccaag agctggaagt tgtgaggcaa cagcgggagc gtctgcagga ggagctaagc 1140
caggcagaga gcaccattga tgagctcaag gagcaggtgg atgctgctct gggtgctgag 1200
gagatggtgg agatgctgac agatcggaac ctgaatctgg aagagaaagt gcgcgagttg 1260
agggagactg tgggagactt ggaagcgatg aatgagatga acgatgagct gcaggagaat 1320
gcacgtgaga cagaactgga gctgcgggag cagctggaca tggcaggcgc gcgggttcgt 1380
gaggcccaga agcgtgtgga ggcagcccag gagacggttg cagactacca gcagaccatc 1440
aagaagtacc gccagctgac cgcccatcta caggatgtga atcgggaact gacaaaccag 1500
caggaagcat ctgtggagag gcaacagcag ccacctccag agacctttga cttcaaaatc 1560
aagtttgctg agactaaggc ccatgccaag gcaattgaga tggaattgag gcagatggag 1620
gtggcccagg ccaatcgaca catgtccctg ctgacagcct tcatgcctga cagcttcctt 1680
cggccaggtg gggaccatga ctgcgttctg gtgctgttgc tcatgcctcg tctcatttgc 1740
aaggcagagc tgatccggaa gcaggcccag gagaagtttg aactaagtga gaactgttca 1800
gagcggcctg ggctgcgagg agctgctggg gagcaactca gctttgctgc tggactggtg 1860
tactcgctga gcctgctgca ggccacgcta caccgctatg agcatgccct ctctcagtgc 1920
agtgtggatg tgtataagaa agtgggcagc ctgtaccctg agatgagtgc ccatgagcgc 1980
tccttggatt tcctcattga actgctgcac aaggatcagc tggatgagac tgtcaatgtg 2040
gagcctctca ccaaggccat caagtactat cagcatctgt acagcatcca ccttgccgaa 2100
cagcctgagg actgtactat gcagctggct gaccacatta agttcacgca gagtgctctg 2160
gactgcatga gtgtggaggt aggacggctg cgtgccttct tgcagggtgg gcaggaggct 2220
acagatattg ccctcctgct ccgggatctg gaaacttcat gcagtgacat ccgccagttc 2280
tgcaagaaga tccgaaggcg aatgccaggg acagatgctc ctgggatccc agctgcactg 2340
gcctttggac cacaggtatc tgacacgctc ctagactgca ggaaacactt gacgtgggtc 2400
gtggctgtgc tgcaggaggt ggcagctgct gctgcccagc tcattgcccc actggcagag 2460
aatgaggggc tacttgtggc tgctctggag gaactggctt tcaaagcaag cgagcagatc 2520
tatgggaccc cctccagcag cccctatgag tgtctgcgcc agtcatgcaa catcctcatc 2580
agtaccatga acaagctggc cacagccatg caggaggggg agtatgatgc agagcggccc 2640
cccagcaagc ctccaccggt tgaactgcgg gctgctgccc ttcgtgcaga gatcacagat 2700
gctgaaggcc tgggtttgaa gctcgaagat cgagagacag ttattaagga gttgaagaag 2760
tcactcaaga ttaagggaga ggagctaagt gaggccaatg tgcggctgag cctcctggag 2820
aagaagttgg acagtgctgc caaggatgca gatgagcgca tcgagaaagt ccagactcgg 2880
ctggaggaga cccaggcact gctgcgaaag aaggagaaag agtttgagga gacaatggat 2940
gcactccagg ctgacatcga ccagctggag gcagagaagg cagaactaaa gcagcgtctg 3000
aacagccagt ccaaacgcac gattgaggga ctccggggcc ctcctccttc aggcattgct 3060
actctggtct ctggcattgc tggtggagcc atccctgggc aggctccagg gtctgtgcca 3120
ggcccagggc tggtgaagga ctcaccactg ctgcttcagc agatctctgc catgaggctg 3180
cacatctccc agctccagca tgagaacagc atcctcaagg gagcccagat gaaggcatcc 3240
ttggcatccc tgccccctct gcatgttgca aagctatccc atgagggccc tggcagtgag 3300
ttaccagctg gagcgctgta tcgtaagacc agccagctgc tggagacatt gaatcaattg 3360
agcacacaca cgcacgtagt agacatcact cgcaccagcc ctgctgccaa gagcccgtcg 3420
gcccaactta tggagcaagt ggctcagctt aagtccctga gtgacaccgt cgagaagctc 3480
aaggatgagg tcctcaagga gacagtatct cagcgccctg gagccacagt acccactgac 3540
tttgccacct tcccttcatc agccttcctc agggaggatc caaagtggga attccctcgg 3600
aagaacttgg ttcttggaaa aactctagga gaaggcgaat ttggaaaagt ggtcaaggca 3660
acggccttcc atctgaaagg cagagcaggg tacaccacgg tggccgtgaa gatgctgaaa 3720
gagaacgcct ccccgagtga gcttcgagac ctgctgtcag agttcaacgt cctgaagcag 3780
gtcaaccacc cacatgtcat caaattgtat ggggcctgca gccaggatgg cccgctcctc 3840
ctcatcgtgg agtacgccaa atacggctcc ctgcggggct tcctccgcga gagccgcaaa 3900
gtggggcctg gctacctggg cagtggaggc agccgcaact ccagctccct ggaccacccg 3960
gatgagcggg ccctcaccat gggcgacctc atctcatttg cctggcagat ctcacagggg 4020
atgcagtatc tggccgagat gaagctcgtt catcgggact tggcagccag aaacatcctg 4080
gtagctgagg ggcggaagat gaagatttcg gatttcggct tgtcccgaga tgtttatgaa 4140
gaggattcct acgtgaagag gagccagggt cggattccag ttaaatggat ggcaattgaa 4200
tccctttttg atcatatcta caccacgcaa agtgatgtat ggtcttttgg tgtcctgctg 4260
tgggagatcg tgaccctagg gggaaacccc tatcctggga ttcctcctga gcggctcttc 4320
aaccttctga agaccggcca ccggatggag aggccagaca actgcagcga ggagatgtac 4380
cgcctgatgc tgcaatgctg gaagcaggag ccggacaaaa ggccggtgtt tgcggacatc 4440
agcaaagacc tggagaagat gatggttaag aggagagact acttggacct tgcggcgtcc 4500
actccatctg actccctgat ttatgacgac ggcctctcag aggaggagac accgctggtg 4560
gactgtaata atgcccccct ccctcgagcc ctcccttcca catggattga aaacaaactc 4620
tatggtagaa tttcccatgc atttactaga ttctag 4656
<210> 20
<211> 1551
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 20
Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg Val
1 5 10 15
Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly Ala
20 25 30
Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu Ala
35 40 45
Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr Cys
50 55 60
Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val Phe
65 70 75 80
Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser Ala
85 90 95
Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr Ser
100 105 110
Lys Leu Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser
115 120 125
Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser
130 135 140
Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro
145 150 155 160
Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro
165 170 175
Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu
180 185 190
Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu
195 200 205
Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp
210 215 220
Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys
225 230 235 240
Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr
245 250 255
Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu
260 265 270
Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val
275 280 285
Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile
290 295 300
Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser
305 310 315 320
Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala
325 330 335
Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val
340 345 350
Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val
355 360 365
Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser
370 375 380
Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu
385 390 395 400
Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys
405 410 415
Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu
420 425 430
Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu
435 440 445
Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys
450 455 460
Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile
465 470 475 480
Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu
485 490 495
Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro
500 505 510
Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His
515 520 525
Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala
530 535 540
Asn Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu
545 550 555 560
Arg Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro
565 570 575
Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys
580 585 590
Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala
595 600 605
Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser
610 615 620
Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys
625 630 635 640
Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser
645 650 655
Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp
660 665 670
Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys
675 680 685
Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp
690 695 700
Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu
705 710 715 720
Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly
725 730 735
Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr
740 745 750
Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met
755 760 765
Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro
770 775 780
Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val
785 790 795 800
Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala
805 810 815
Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu
820 825 830
Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro
835 840 845
Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn
850 855 860
Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro
865 870 875 880
Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala
885 890 895
Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu
900 905 910
Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu
915 920 925
Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp
930 935 940
Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg
945 950 955 960
Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu
965 970 975
Glu Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu
980 985 990
Lys Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile
995 1000 1005
Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val
1010 1015 1020
Ser Gly Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser
1025 1030 1035
Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln
1040 1045 1050
Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu
1055 1060 1065
Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser
1070 1075 1080
Leu Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly
1085 1090 1095
Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu
1100 1105 1110
Leu Glu Thr Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp
1115 1120 1125
Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu
1130 1135 1140
Met Glu Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu
1145 1150 1155
Lys Leu Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro
1160 1165 1170
Gly Ala Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala
1175 1180 1185
Phe Leu Arg Glu Asp Pro Lys Trp Glu Phe Pro Arg Lys Asn Leu
1190 1195 1200
Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly Lys Val Val
1205 1210 1215
Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr Thr Thr
1220 1225 1230
Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu Leu
1235 1240 1245
Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His
1250 1255 1260
Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro
1265 1270 1275
Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly
1280 1285 1290
Phe Leu Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser
1295 1300 1305
Gly Gly Ser Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg
1310 1315 1320
Ala Leu Thr Met Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser
1325 1330 1335
Gln Gly Met Gln Tyr Leu Ala Glu Met Lys Leu Val His Arg Asp
1340 1345 1350
Leu Ala Ala Arg Asn Ile Leu Val Ala Glu Gly Arg Lys Met Lys
1355 1360 1365
Ile Ser Asp Phe Gly Leu Ser Arg Asp Val Tyr Glu Glu Asp Ser
1370 1375 1380
Tyr Val Lys Arg Ser Gln Gly Arg Ile Pro Val Lys Trp Met Ala
1385 1390 1395
Ile Glu Ser Leu Phe Asp His Ile Tyr Thr Thr Gln Ser Asp Val
1400 1405 1410
Trp Ser Phe Gly Val Leu Leu Trp Glu Ile Val Thr Leu Gly Gly
1415 1420 1425
Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu Phe Asn Leu Leu
1430 1435 1440
Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys Ser Glu Glu
1445 1450 1455
Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro Asp Lys
1460 1465 1470
Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met Met
1475 1480 1485
Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro Ser
1490 1495 1500
Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr Pro
1505 1510 1515
Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro Ser
1520 1525 1530
Thr Trp Ile Glu Asn Lys Leu Tyr Gly Arg Ile Ser His Ala Phe
1535 1540 1545
Thr Arg Phe
1550
<210> 21
<211> 4887
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 21
atggcacaga gcaagaggca cgtgtacagc cggacgccca gcggcagcag gatgagtgcg 60
gaggcaagcg cccggcctct gcgggtgggc tcccgtgtag aggtgattgg aaaaggccac 120
cgaggcactg tggcctatgt tggagccaca ctgtttgcca ctggcaaatg ggtaggcgtg 180
attctggatg aagcaaaggg caaaaatgat ggaactgttc aaggcaggaa gtacttcact 240
tgtgatgaag ggcatggcat ctttgtgcgc cagtcccaga tccaggtatt tgaagatgga 300
gcagatacta cttccccaga gacacctgat tcttctgctt caaaagtcct caaaagagag 360
ggaactgata caactgcaaa gactagcaaa ctggcaccga cagcccgaaa gaccacaact 420
cggcgaccca agcccacgcg cccagccagt actggggtgg ctggggccag tagctccctg 480
ggcccctctg gctcagcgtc agcaggtgag ctgagcagca gtgagcccag caccccggct 540
cagactccgc tggcagcacc catcatcccc acgccggtcc tcacctctcc tggagcagtc 600
cccccgcttc cttccccatc caaggaggag gagggactaa gggctcaggt gcgggacctg 660
gaggagaaac tagagaccct gagactgaaa cgggcagaag acaaagcaaa gctaaaagag 720
ctggagaaac acaaaatcca gctggagcag gtgcaggaat ggaagagcaa aatgcaggag 780
cagcaggccg acctgcagcg gcgcctcaag gaggcgagaa aggaagccaa ggaggcgctg 840
gaggcaaagg aacgctatat ggaggagatg gctgatactg ctgatgccat tgagatggcc 900
actttggaca aggagatggc tgaagagcgg gctgagtccc tgcagcagga ggtggaggca 960
ctgaaggagc gggtggacga gctcactact gacttagaga tcctcaaggc tgagattgaa 1020
gagaagggct cagatggcgc tgcatccagt tatcagctca agcagcttga ggagcagaat 1080
gcccgcctga aggatgccct ggtgaggatg cgggatcttt cttcctcaga gaagcaggag 1140
catgtgaagc tccagaagct catggaaaag aagaaccaag agctggaagt tgtgaggcaa 1200
cagcgggagc gtctgcagga ggagctaagc caggcagaga gcaccattga tgagctcaag 1260
gagcaggtgg atgctgctct gggtgctgag gagatggtgg agatgctgac agatcggaac 1320
ctgaatctgg aagagaaagt gcgcgagttg agggagactg tgggagactt ggaagcgatg 1380
aatgagatga acgatgagct gcaggagaat gcacgtgaga cagaactgga gctgcgggag 1440
cagctggaca tggcaggcgc gcgggttcgt gaggcccaga agcgtgtgga ggcagcccag 1500
gagacggttg cagactacca gcagaccatc aagaagtacc gccagctgac cgcccatcta 1560
caggatgtga atcgggaact gacaaaccag caggaagcat ctgtggagag gcaacagcag 1620
ccacctccag agacctttga cttcaaaatc aagtttgctg agactaaggc ccatgccaag 1680
gcaattgaga tggaattgag gcagatggag gtggcccagg ccaatcgaca catgtccctg 1740
ctgacagcct tcatgcctga cagcttcctt cggccaggtg gggaccatga ctgcgttctg 1800
gtgctgttgc tcatgcctcg tctcatttgc aaggcagagc tgatccggaa gcaggcccag 1860
gagaagtttg aactaagtga gaactgttca gagcggcctg ggctgcgagg agctgctggg 1920
gagcaactca gctttgctgc tggactggtg tactcgctga gcctgctgca ggccacgcta 1980
caccgctatg agcatgccct ctctcagtgc agtgtggatg tgtataagaa agtgggcagc 2040
ctgtaccctg agatgagtgc ccatgagcgc tccttggatt tcctcattga actgctgcac 2100
aaggatcagc tggatgagac tgtcaatgtg gagcctctca ccaaggccat caagtactat 2160
cagcatctgt acagcatcca ccttgccgaa cagcctgagg actgtactat gcagctggct 2220
gaccacatta agttcacgca gagtgctctg gactgcatga gtgtggaggt aggacggctg 2280
cgtgccttct tgcagggtgg gcaggaggct acagatattg ccctcctgct ccgggatctg 2340
gaaacttcat gcagtgacat ccgccagttc tgcaagaaga tccgaaggcg aatgccaggg 2400
acagatgctc ctgggatccc agctgcactg gcctttggac cacaggtatc tgacacgctc 2460
ctagactgca ggaaacactt gacgtgggtc gtggctgtgc tgcaggaggt ggcagctgct 2520
gctgcccagc tcattgcccc actggcagag aatgaggggc tacttgtggc tgctctggag 2580
gaactggctt tcaaagcaag cgagcagatc tatgggaccc cctccagcag cccctatgag 2640
tgtctgcgcc agtcatgcaa catcctcatc agtaccatga acaagctggc cacagccatg 2700
caggaggggg agtatgatgc agagcggccc cccagcaagc ctccaccggt tgaactgcgg 2760
gctgctgccc ttcgtgcaga gatcacagat gctgaaggcc tgggtttgaa gctcgaagat 2820
cgagagacag ttattaagga gttgaagaag tcactcaaga ttaagggaga ggagctaagt 2880
gaggccaatg tgcggctgag cctcctggag aagaagttgg acagtgctgc caaggatgca 2940
gatgagcgca tcgagaaagt ccagactcgg ctggaggaga cccaggcact gctgcgaaag 3000
aaggagaaag agtttgagga gacaatggat gcactccagg ctgacatcga ccagctggag 3060
gcagagaagg cagaactaaa gcagcgtctg aacagccagt ccaaacgcac gattgaggga 3120
ctccggggcc ctcctccttc aggcattgct actctggtct ctggcattgc tggtgaagaa 3180
cagcagcgag gagccatccc tgggcaggct ccagggtctg tgccaggccc agggctggtg 3240
aaggactcac cactgctgct tcagcagatc tctgccatga ggctgcacat ctcccagctc 3300
cagcatgaga acagcatcct caagggagcc cagatgaagg catccttggc atccctgccc 3360
cctctgcatg ttgcaaagct atcccatgag ggccctggca gtgagttacc agctggagcg 3420
ctgtatcgta agaccagcca gctgctggag acattgaatc aattgagcac acacacgcac 3480
gtagtagaca tcactcgcac cagccctgct gccaagagcc cgtcggccca acttatggag 3540
caagtggctc agcttaagtc cctgagtgac accgtcgaga agctcaagga tgaggtcctc 3600
aaggagacag tatctcagcg ccctggagcc acagtaccca ctgactttgc caccttccct 3660
tcatcagcct tcctcaggga ggatccaaag tgggaattcc ctcggaagaa cttggttctt 3720
ggaaaaactc taggagaagg cgaatttgga aaagtggtca aggcaacggc cttccatctg 3780
aaaggcagag cagggtacac cacggtggcc gtgaagatgc tgaaagagaa cgcctccccg 3840
agtgagcttc gagacctgct gtcagagttc aacgtcctga agcaggtcaa ccacccacat 3900
gtcatcaaat tgtatggggc ctgcagccag gatggcccgc tcctcctcat cgtggagtac 3960
gccaaatacg gctccctgcg gggcttcctc cgcgagagcc gcaaagtggg gcctggctac 4020
ctgggcagtg gaggcagccg caactccagc tccctggacc acccggatga gcgggccctc 4080
accatgggcg acctcatctc atttgcctgg cagatctcac aggggatgca gtatctggcc 4140
gagatgaagc tcgttcatcg ggacttggca gccagaaaca tcctggtagc tgaggggcgg 4200
aagatgaaga tttcggattt cggcttgtcc cgagatgttt atgaagagga ttcctacgtg 4260
aagaggagcc agggtcggat tccagttaaa tggatggcaa ttgaatccct ttttgatcat 4320
atctacacca cgcaaagtga tgtatggtct tttggtgtcc tgctgtggga gatcgtgacc 4380
ctagggggaa acccctatcc tgggattcct cctgagcggc tcttcaacct tctgaagacc 4440
ggccaccgga tggagaggcc agacaactgc agcgaggaga tgtaccgcct gatgctgcaa 4500
tgctggaagc aggagccgga caaaaggccg gtgtttgcgg acatcagcaa agacctggag 4560
aagatgatgg ttaagaggag agactacttg gaccttgcgg cgtccactcc atctgactcc 4620
ctgatttatg acgacggcct ctcagaggag gagacaccgc tggtggactg taataatgcc 4680
cccctccctc gagccctccc ttccacatgg attgaaaaca aactctatgg catgtcagac 4740
ccgaactggc ctggagagag tcctgtacca ctcacgagag ctgatggcac taacactggg 4800
tttccaagat atccaaatga tagtgtatat gctaactgga tgctttcacc ctcagcggca 4860
aaattaatgg acacgtttga tagttaa 4887
<210> 22
<211> 1628
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 22
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro Lys
130 135 140
Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser Leu
145 150 155 160
Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu Pro
165 170 175
Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr Pro
180 185 190
Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser Lys
195 200 205
Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys Leu
210 215 220
Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys Glu
225 230 235 240
Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys Ser
245 250 255
Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu Ala
260 265 270
Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met Glu
275 280 285
Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp Lys
290 295 300
Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu Ala
305 310 315 320
Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu Lys
325 330 335
Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr Gln
340 345 350
Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu Val
355 360 365
Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys Leu
370 375 380
Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg Gln
385 390 395 400
Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr Ile
405 410 415
Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu Met
420 425 430
Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val Arg
435 440 445
Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met Asn
450 455 460
Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg Glu
465 470 475 480
Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg Val
485 490 495
Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys Lys
500 505 510
Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu Thr
515 520 525
Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro Glu
530 535 540
Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala Lys
545 550 555 560
Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn Arg
565 570 575
His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg Pro
580 585 590
Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg Leu
595 600 605
Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe Glu
610 615 620
Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala Gly
625 630 635 640
Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu Leu
645 650 655
Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser Val
660 665 670
Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala His
675 680 685
Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln Leu
690 695 700
Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr Tyr
705 710 715 720
Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys Thr
725 730 735
Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp Cys
740 745 750
Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly Gln
755 760 765
Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser Cys
770 775 780
Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro Gly
785 790 795 800
Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln Val
805 810 815
Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val Ala
820 825 830
Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro Leu
835 840 845
Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala Phe
850 855 860
Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr Glu
865 870 875 880
Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys Leu
885 890 895
Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro Ser
900 905 910
Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu Ile
915 920 925
Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr Val
930 935 940
Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu Ser
945 950 955 960
Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser Ala
965 970 975
Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu Glu
980 985 990
Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu Thr
995 1000 1005
Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
1010 1015 1020
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile
1025 1030 1035
Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val
1040 1045 1050
Ser Gly Ile Ala Gly Glu Glu Gln Gln Arg Gly Ala Ile Pro Gly
1055 1060 1065
Gln Ala Pro Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser
1070 1075 1080
Pro Leu Leu Leu Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser
1085 1090 1095
Gln Leu Gln His Glu Asn Ser Ile Leu Lys Gly Ala Gln Met Lys
1100 1105 1110
Ala Ser Leu Ala Ser Leu Pro Pro Leu His Val Ala Lys Leu Ser
1115 1120 1125
His Glu Gly Pro Gly Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg
1130 1135 1140
Lys Thr Ser Gln Leu Leu Glu Thr Leu Asn Gln Leu Ser Thr His
1145 1150 1155
Thr His Val Val Asp Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser
1160 1165 1170
Pro Ser Ala Gln Leu Met Glu Gln Val Ala Gln Leu Lys Ser Leu
1175 1180 1185
Ser Asp Thr Val Glu Lys Leu Lys Asp Glu Val Leu Lys Glu Thr
1190 1195 1200
Val Ser Gln Arg Pro Gly Ala Thr Val Pro Thr Asp Phe Ala Thr
1205 1210 1215
Phe Pro Ser Ser Ala Phe Leu Arg Glu Asp Pro Lys Trp Glu Phe
1220 1225 1230
Pro Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu
1235 1240 1245
Phe Gly Lys Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg
1250 1255 1260
Ala Gly Tyr Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala
1265 1270 1275
Ser Pro Ser Glu Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu
1280 1285 1290
Lys Gln Val Asn His Pro His Val Ile Lys Leu Tyr Gly Ala Cys
1295 1300 1305
Ser Gln Asp Gly Pro Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr
1310 1315 1320
Gly Ser Leu Arg Gly Phe Leu Arg Glu Ser Arg Lys Val Gly Pro
1325 1330 1335
Gly Tyr Leu Gly Ser Gly Gly Ser Arg Asn Ser Ser Ser Leu Asp
1340 1345 1350
His Pro Asp Glu Arg Ala Leu Thr Met Gly Asp Leu Ile Ser Phe
1355 1360 1365
Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr Leu Ala Glu Met Lys
1370 1375 1380
Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Ala Glu
1385 1390 1395
Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser Arg Asp Val
1400 1405 1410
Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg Ile Pro
1415 1420 1425
Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr Thr
1430 1435 1440
Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile
1445 1450 1455
Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg
1460 1465 1470
Leu Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp
1475 1480 1485
Asn Cys Ser Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys
1490 1495 1500
Gln Glu Pro Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp
1505 1510 1515
Leu Glu Lys Met Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala
1520 1525 1530
Ala Ser Thr Pro Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser
1535 1540 1545
Glu Glu Glu Thr Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro
1550 1555 1560
Arg Ala Leu Pro Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Met
1565 1570 1575
Ser Asp Pro Asn Trp Pro Gly Glu Ser Pro Val Pro Leu Thr Arg
1580 1585 1590
Ala Asp Gly Thr Asn Thr Gly Phe Pro Arg Tyr Pro Asn Asp Ser
1595 1600 1605
Val Tyr Ala Asn Trp Met Leu Ser Pro Ser Ala Ala Lys Leu Met
1610 1615 1620
Asp Thr Phe Asp Ser
1625
<210> 23
<211> 4761
<212> DNA
<213> 人工序列
<220>
<223> 融合肽的编码序列
<400> 23
atggcacaga gcaagaggca cgtgtacagc cggacgccca gcggcagcag gatgagtgcg 60
gaggcaagcg cccggcctct gcgggtgggc tcccgtgtag aggtgattgg aaaaggccac 120
cgaggcactg tggcctatgt tggagccaca ctgtttgcca ctggcaaatg ggtaggcgtg 180
attctggatg aagcaaaggg caaaaatgat ggaactgttc aaggcaggaa gtacttcact 240
tgtgatgaag ggcatggcat ctttgtgcgc cagtcccaga tccaggtatt tgaagatgga 300
gcagatacta cttccccaga gacacctgat tcttctgctt caaaagtcct caaaagagag 360
ggaactgata caactgcaaa gactagcaaa ctggcaccga cagcccgaaa gaccacaact 420
cggcgaccca agcccacgcg cccagccagt actggggtgg ctggggccag tagctccctg 480
ggcccctctg gctcagcgtc agcaggtgag ctgagcagca gtgagcccag caccccggct 540
cagactccgc tggcagcacc catcatcccc acgccggtcc tcacctctcc tggagcagtc 600
cccccgcttc cttccccatc caaggaggag gagggactaa gggctcaggt gcgggacctg 660
gaggagaaac tagagaccct gagactgaaa cgggcagaag acaaagcaaa gctaaaagag 720
ctggagaaac acaaaatcca gctggagcag gtgcaggaat ggaagagcaa aatgcaggag 780
cagcaggccg acctgcagcg gcgcctcaag gaggcgagaa aggaagccaa ggaggcgctg 840
gaggcaaagg aacgctatat ggaggagatg gctgatactg ctgatgccat tgagatggcc 900
actttggaca aggagatggc tgaagagcgg gctgagtccc tgcagcagga ggtggaggca 960
ctgaaggagc gggtggacga gctcactact gacttagaga tcctcaaggc tgagattgaa 1020
gagaagggct cagatggcgc tgcatccagt tatcagctca agcagcttga ggagcagaat 1080
gcccgcctga aggatgccct ggtgaggatg cgggatcttt cttcctcaga gaagcaggag 1140
catgtgaagc tccagaagct catggaaaag aagaaccaag agctggaagt tgtgaggcaa 1200
cagcgggagc gtctgcagga ggagctaagc caggcagaga gcaccattga tgagctcaag 1260
gagcaggtgg atgctgctct gggtgctgag gagatggtgg agatgctgac agatcggaac 1320
ctgaatctgg aagagaaagt gcgcgagttg agggagactg tgggagactt ggaagcgatg 1380
aatgagatga acgatgagct gcaggagaat gcacgtgaga cagaactgga gctgcgggag 1440
cagctggaca tggcaggcgc gcgggttcgt gaggcccaga agcgtgtgga ggcagcccag 1500
gagacggttg cagactacca gcagaccatc aagaagtacc gccagctgac cgcccatcta 1560
caggatgtga atcgggaact gacaaaccag caggaagcat ctgtggagag gcaacagcag 1620
ccacctccag agacctttga cttcaaaatc aagtttgctg agactaaggc ccatgccaag 1680
gcaattgaga tggaattgag gcagatggag gtggcccagg ccaatcgaca catgtccctg 1740
ctgacagcct tcatgcctga cagcttcctt cggccaggtg gggaccatga ctgcgttctg 1800
gtgctgttgc tcatgcctcg tctcatttgc aaggcagagc tgatccggaa gcaggcccag 1860
gagaagtttg aactaagtga gaactgttca gagcggcctg ggctgcgagg agctgctggg 1920
gagcaactca gctttgctgc tggactggtg tactcgctga gcctgctgca ggccacgcta 1980
caccgctatg agcatgccct ctctcagtgc agtgtggatg tgtataagaa agtgggcagc 2040
ctgtaccctg agatgagtgc ccatgagcgc tccttggatt tcctcattga actgctgcac 2100
aaggatcagc tggatgagac tgtcaatgtg gagcctctca ccaaggccat caagtactat 2160
cagcatctgt acagcatcca ccttgccgaa cagcctgagg actgtactat gcagctggct 2220
gaccacatta agttcacgca gagtgctctg gactgcatga gtgtggaggt aggacggctg 2280
cgtgccttct tgcagggtgg gcaggaggct acagatattg ccctcctgct ccgggatctg 2340
gaaacttcat gcagtgacat ccgccagttc tgcaagaaga tccgaaggcg aatgccaggg 2400
acagatgctc ctgggatccc agctgcactg gcctttggac cacaggtatc tgacacgctc 2460
ctagactgca ggaaacactt gacgtgggtc gtggctgtgc tgcaggaggt ggcagctgct 2520
gctgcccagc tcattgcccc actggcagag aatgaggggc tacttgtggc tgctctggag 2580
gaactggctt tcaaagcaag cgagcagatc tatgggaccc cctccagcag cccctatgag 2640
tgtctgcgcc agtcatgcaa catcctcatc agtaccatga acaagctggc cacagccatg 2700
caggaggggg agtatgatgc agagcggccc cccagcaagc ctccaccggt tgaactgcgg 2760
gctgctgccc ttcgtgcaga gatcacagat gctgaaggcc tgggtttgaa gctcgaagat 2820
cgagagacag ttattaagga gttgaagaag tcactcaaga ttaagggaga ggagctaagt 2880
gaggccaatg tgcggctgag cctcctggag aagaagttgg acagtgctgc caaggatgca 2940
gatgagcgca tcgagaaagt ccagactcgg ctggaggaga cccaggcact gctgcgaaag 3000
aaggagaaag agtttgagga gacaatggat gcactccagg ctgacatcga ccagctggag 3060
gcagagaagg cagaactaaa gcagcgtctg aacagccagt ccaaacgcac gattgaggga 3120
ctccggggcc ctcctccttc aggcattgct actctggtct ctggcattgc tggtgaagaa 3180
cagcagcgag gagccatccc tgggcaggct ccagggtctg tgccaggccc agggctggtg 3240
aaggactcac cactgctgct tcagcagatc tctgccatga ggctgcacat ctcccagctc 3300
cagcatgaga acagcatcct caagggagcc cagatgaagg catccttggc atccctgccc 3360
cctctgcatg ttgcaaagct atcccatgag ggccctggca gtgagttacc agctggagcg 3420
ctgtatcgta agaccagcca gctgctggag acattgaatc aattgagcac acacacgcac 3480
gtagtagaca tcactcgcac cagccctgct gccaagagcc cgtcggccca acttatggag 3540
caagtggctc agcttaagtc cctgagtgac accgtcgaga agctcaagga tgaggtcctc 3600
aaggagacag tatctcagcg ccctggagcc acagtaccca ctgactttgc caccttccct 3660
tcatcagcct tcctcaggga ggatccaaag tgggaattcc ctcggaagaa cttggttctt 3720
ggaaaaactc taggagaagg cgaatttgga aaagtggtca aggcaacggc cttccatctg 3780
aaaggcagag cagggtacac cacggtggcc gtgaagatgc tgaaagagaa cgcctccccg 3840
agtgagcttc gagacctgct gtcagagttc aacgtcctga agcaggtcaa ccacccacat 3900
gtcatcaaat tgtatggggc ctgcagccag gatggcccgc tcctcctcat cgtggagtac 3960
gccaaatacg gctccctgcg gggcttcctc cgcgagagcc gcaaagtggg gcctggctac 4020
ctgggcagtg gaggcagccg caactccagc tccctggacc acccggatga gcgggccctc 4080
accatgggcg acctcatctc atttgcctgg cagatctcac aggggatgca gtatctggcc 4140
gagatgaagc tcgttcatcg ggacttggca gccagaaaca tcctggtagc tgaggggcgg 4200
aagatgaaga tttcggattt cggcttgtcc cgagatgttt atgaagagga ttcctacgtg 4260
aagaggagcc agggtcggat tccagttaaa tggatggcaa ttgaatccct ttttgatcat 4320
atctacacca cgcaaagtga tgtatggtct tttggtgtcc tgctgtggga gatcgtgacc 4380
ctagggggaa acccctatcc tgggattcct cctgagcggc tcttcaacct tctgaagacc 4440
ggccaccgga tggagaggcc agacaactgc agcgaggaga tgtaccgcct gatgctgcaa 4500
tgctggaagc aggagccgga caaaaggccg gtgtttgcgg acatcagcaa agacctggag 4560
aagatgatgg ttaagaggag agactacttg gaccttgcgg cgtccactcc atctgactcc 4620
ctgatttatg acgacggcct ctcagaggag gagacaccgc tggtggactg taataatgcc 4680
cccctccctc gagccctccc ttccacatgg attgaaaaca aactctatgg tagaatttcc 4740
catgcattta ctagattcta g 4761
<210> 24
<211> 1586
<212> PRT
<213> 人工序列
<220>
<223> 融合肽
<400> 24
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro Lys
130 135 140
Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser Leu
145 150 155 160
Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu Pro
165 170 175
Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr Pro
180 185 190
Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser Lys
195 200 205
Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys Leu
210 215 220
Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys Glu
225 230 235 240
Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys Ser
245 250 255
Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu Ala
260 265 270
Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met Glu
275 280 285
Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp Lys
290 295 300
Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu Ala
305 310 315 320
Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu Lys
325 330 335
Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr Gln
340 345 350
Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu Val
355 360 365
Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys Leu
370 375 380
Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg Gln
385 390 395 400
Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr Ile
405 410 415
Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu Met
420 425 430
Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val Arg
435 440 445
Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met Asn
450 455 460
Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg Glu
465 470 475 480
Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg Val
485 490 495
Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys Lys
500 505 510
Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu Thr
515 520 525
Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro Glu
530 535 540
Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala Lys
545 550 555 560
Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn Arg
565 570 575
His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg Pro
580 585 590
Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg Leu
595 600 605
Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe Glu
610 615 620
Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala Gly
625 630 635 640
Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu Leu
645 650 655
Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser Val
660 665 670
Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala His
675 680 685
Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln Leu
690 695 700
Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr Tyr
705 710 715 720
Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys Thr
725 730 735
Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp Cys
740 745 750
Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly Gln
755 760 765
Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser Cys
770 775 780
Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro Gly
785 790 795 800
Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln Val
805 810 815
Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val Ala
820 825 830
Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro Leu
835 840 845
Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala Phe
850 855 860
Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr Glu
865 870 875 880
Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys Leu
885 890 895
Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro Ser
900 905 910
Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu Ile
915 920 925
Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr Val
930 935 940
Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu Ser
945 950 955 960
Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser Ala
965 970 975
Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu Glu
980 985 990
Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu Thr
995 1000 1005
Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
1010 1015 1020
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile
1025 1030 1035
Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val
1040 1045 1050
Ser Gly Ile Ala Gly Glu Glu Gln Gln Arg Gly Ala Ile Pro Gly
1055 1060 1065
Gln Ala Pro Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser
1070 1075 1080
Pro Leu Leu Leu Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser
1085 1090 1095
Gln Leu Gln His Glu Asn Ser Ile Leu Lys Gly Ala Gln Met Lys
1100 1105 1110
Ala Ser Leu Ala Ser Leu Pro Pro Leu His Val Ala Lys Leu Ser
1115 1120 1125
His Glu Gly Pro Gly Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg
1130 1135 1140
Lys Thr Ser Gln Leu Leu Glu Thr Leu Asn Gln Leu Ser Thr His
1145 1150 1155
Thr His Val Val Asp Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser
1160 1165 1170
Pro Ser Ala Gln Leu Met Glu Gln Val Ala Gln Leu Lys Ser Leu
1175 1180 1185
Ser Asp Thr Val Glu Lys Leu Lys Asp Glu Val Leu Lys Glu Thr
1190 1195 1200
Val Ser Gln Arg Pro Gly Ala Thr Val Pro Thr Asp Phe Ala Thr
1205 1210 1215
Phe Pro Ser Ser Ala Phe Leu Arg Glu Asp Pro Lys Trp Glu Phe
1220 1225 1230
Pro Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu
1235 1240 1245
Phe Gly Lys Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg
1250 1255 1260
Ala Gly Tyr Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala
1265 1270 1275
Ser Pro Ser Glu Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu
1280 1285 1290
Lys Gln Val Asn His Pro His Val Ile Lys Leu Tyr Gly Ala Cys
1295 1300 1305
Ser Gln Asp Gly Pro Leu Leu Leu Ile Val Glu Tyr Ala Lys Tyr
1310 1315 1320
Gly Ser Leu Arg Gly Phe Leu Arg Glu Ser Arg Lys Val Gly Pro
1325 1330 1335
Gly Tyr Leu Gly Ser Gly Gly Ser Arg Asn Ser Ser Ser Leu Asp
1340 1345 1350
His Pro Asp Glu Arg Ala Leu Thr Met Gly Asp Leu Ile Ser Phe
1355 1360 1365
Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr Leu Ala Glu Met Lys
1370 1375 1380
Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Ala Glu
1385 1390 1395
Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser Arg Asp Val
1400 1405 1410
Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg Ile Pro
1415 1420 1425
Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr Thr
1430 1435 1440
Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile
1445 1450 1455
Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg
1460 1465 1470
Leu Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp
1475 1480 1485
Asn Cys Ser Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys
1490 1495 1500
Gln Glu Pro Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp
1505 1510 1515
Leu Glu Lys Met Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala
1520 1525 1530
Ala Ser Thr Pro Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser
1535 1540 1545
Glu Glu Glu Thr Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro
1550 1555 1560
Arg Ala Leu Pro Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Arg
1565 1570 1575
Ile Ser His Ala Phe Thr Arg Phe
1580 1585
<210> 25
<211> 1278
<212> PRT
<213> 智人
<400> 25
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Arg Gly Leu Lys Pro Lys Lys Ala Pro Thr Ala Arg Lys
130 135 140
Thr Thr Thr Arg Arg Pro Lys Pro Thr Arg Pro Ala Ser Thr Gly Val
145 150 155 160
Ala Gly Ala Ser Ser Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly
165 170 175
Glu Leu Ser Ser Ser Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala
180 185 190
Ala Pro Ile Ile Pro Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro
195 200 205
Pro Leu Pro Ser Pro Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val
210 215 220
Arg Asp Leu Glu Glu Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu
225 230 235 240
Asp Lys Ala Lys Leu Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu
245 250 255
Gln Val Gln Glu Trp Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu
260 265 270
Gln Arg Arg Leu Lys Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu
275 280 285
Ala Lys Glu Arg Tyr Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile
290 295 300
Glu Met Ala Thr Leu Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser
305 310 315 320
Leu Gln Gln Glu Val Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr
325 330 335
Thr Asp Leu Glu Ile Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp
340 345 350
Gly Ala Ala Ser Ser Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala
355 360 365
Arg Leu Lys Asp Ala Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu
370 375 380
Lys Gln Glu His Val Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln
385 390 395 400
Glu Leu Glu Val Val Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu
405 410 415
Ser Gln Ala Glu Ser Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala
420 425 430
Ala Leu Gly Ala Glu Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu
435 440 445
Asn Leu Glu Glu Lys Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu
450 455 460
Glu Ala Met Asn Glu Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu
465 470 475 480
Thr Glu Leu Glu Leu Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val
485 490 495
Arg Glu Ala Gln Lys Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp
500 505 510
Tyr Gln Gln Thr Ile Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln
515 520 525
Asp Val Asn Arg Glu Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg
530 535 540
Gln Gln Gln Pro Pro Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala
545 550 555 560
Glu Thr Lys Ala His Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met
565 570 575
Glu Val Ala Gln Ala Asn Arg His Met Ser Leu Leu Thr Ala Phe Met
580 585 590
Pro Asp Ser Phe Leu Arg Pro Gly Gly Asp His Asp Cys Val Leu Val
595 600 605
Leu Leu Leu Met Pro Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys
610 615 620
Gln Ala Gln Glu Lys Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro
625 630 635 640
Gly Leu Arg Gly Ala Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu
645 650 655
Val Tyr Ser Leu Ser Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His
660 665 670
Ala Leu Ser Gln Cys Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu
675 680 685
Tyr Pro Glu Met Ser Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu
690 695 700
Leu Leu His Lys Asp Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu
705 710 715 720
Thr Lys Ala Ile Lys Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala
725 730 735
Glu Gln Pro Glu Asp Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe
740 745 750
Thr Gln Ser Ala Leu Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg
755 760 765
Ala Phe Leu Gln Gly Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu
770 775 780
Arg Asp Leu Glu Thr Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys
785 790 795 800
Ile Arg Arg Arg Met Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala
805 810 815
Leu Ala Phe Gly Pro Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys
820 825 830
His Leu Thr Trp Val Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala
835 840 845
Ala Gln Leu Ile Ala Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala
850 855 860
Ala Leu Glu Glu Leu Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr
865 870 875 880
Pro Ser Ser Ser Pro Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu
885 890 895
Ile Ser Thr Met Asn Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr
900 905 910
Asp Ala Glu Arg Pro Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala
915 920 925
Ala Ala Leu Arg Ala Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys
930 935 940
Leu Glu Asp Arg Glu Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys
945 950 955 960
Ile Lys Gly Glu Glu Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu
965 970 975
Glu Lys Lys Leu Asp Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu
980 985 990
Lys Val Gln Thr Arg Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys
995 1000 1005
Glu Lys Glu Phe Glu Glu Thr Met Asp Ala Leu Gln Ala Asp Ile
1010 1015 1020
Asp Gln Leu Glu Ala Glu Lys Ala Glu Leu Lys Gln Arg Leu Asn
1025 1030 1035
Ser Gln Ser Lys Arg Thr Ile Glu Gly Leu Arg Gly Pro Pro Pro
1040 1045 1050
Ser Gly Ile Ala Thr Leu Val Ser Gly Ile Ala Gly Glu Glu Gln
1055 1060 1065
Gln Arg Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser Val Pro Gly
1070 1075 1080
Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln Gln Ile Ser
1085 1090 1095
Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu Asn Ser Ile
1100 1105 1110
Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu Pro Pro
1115 1120 1125
Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu Leu
1130 1135 1140
Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu Thr
1145 1150 1155
Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp Ile Thr Arg
1160 1165 1170
Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu Met Glu Gln
1175 1180 1185
Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu Lys Leu Lys
1190 1195 1200
Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro Gly Ala Thr
1205 1210 1215
Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu Arg
1220 1225 1230
Ala Lys Glu Glu Gln Gln Asp Asp Thr Val Tyr Met Gly Lys Val
1235 1240 1245
Thr Phe Ser Cys Ala Ala Gly Phe Gly Gln Arg His Arg Leu Val
1250 1255 1260
Leu Thr Gln Glu Gln Leu His Gln Leu His Ser Arg Leu Ile Ser
1265 1270 1275
<210> 26
<211> 1144
<212> PRT
<213> 智人
<400> 26
Met Met Arg Gln Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro
1 5 10 15
Lys Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser
20 25 30
Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu
35 40 45
Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr
50 55 60
Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser
65 70 75 80
Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys
85 90 95
Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys
100 105 110
Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys
115 120 125
Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu
130 135 140
Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met
145 150 155 160
Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp
165 170 175
Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu
180 185 190
Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu
195 200 205
Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr
210 215 220
Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu
225 230 235 240
Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys
245 250 255
Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg
260 265 270
Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr
275 280 285
Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu
290 295 300
Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val
305 310 315 320
Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met
325 330 335
Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg
340 345 350
Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg
355 360 365
Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys
370 375 380
Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu
385 390 395 400
Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro
405 410 415
Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala
420 425 430
Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn
435 440 445
Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg
450 455 460
Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg
465 470 475 480
Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe
485 490 495
Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala
500 505 510
Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu
515 520 525
Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser
530 535 540
Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala
545 550 555 560
His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln
565 570 575
Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr
580 585 590
Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys
595 600 605
Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp
610 615 620
Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly
625 630 635 640
Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser
645 650 655
Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro
660 665 670
Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln
675 680 685
Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val
690 695 700
Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro
705 710 715 720
Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala
725 730 735
Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr
740 745 750
Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys
755 760 765
Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro
770 775 780
Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu
785 790 795 800
Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr
805 810 815
Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu
820 825 830
Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser
835 840 845
Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu
850 855 860
Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu
865 870 875 880
Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
885 890 895
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile Glu
900 905 910
Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val Ser Gly
915 920 925
Ile Ala Gly Glu Glu Gln Gln Arg Gly Ala Ile Pro Gly Gln Ala Pro
930 935 940
Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu
945 950 955 960
Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu
965 970 975
Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu
980 985 990
Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu
995 1000 1005
Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu
1010 1015 1020
Thr Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp Ile Thr
1025 1030 1035
Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu Met Glu
1040 1045 1050
Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu Lys Leu
1055 1060 1065
Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro Gly Ala
1070 1075 1080
Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu
1085 1090 1095
Arg Ala Lys Glu Glu Gln Gln Asp Asp Thr Val Tyr Met Gly Lys
1100 1105 1110
Val Thr Phe Ser Cys Ala Ala Gly Phe Gly Gln Arg His Arg Leu
1115 1120 1125
Val Leu Thr Gln Glu Gln Leu His Gln Leu His Ser Arg Leu Ile
1130 1135 1140
Ser
<210> 27
<211> 1253
<212> PRT
<213> 智人
<400> 27
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser
130 135 140
Ser Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser
145 150 155 160
Ser Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile
165 170 175
Pro Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser
180 185 190
Pro Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu
195 200 205
Glu Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys
210 215 220
Leu Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu
225 230 235 240
Trp Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu
245 250 255
Lys Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg
260 265 270
Tyr Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr
275 280 285
Leu Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu
290 295 300
Val Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu
305 310 315 320
Ile Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser
325 330 335
Ser Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp
340 345 350
Ala Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His
355 360 365
Val Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val
370 375 380
Val Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu
385 390 395 400
Ser Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala
405 410 415
Glu Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu
420 425 430
Lys Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn
435 440 445
Glu Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu
450 455 460
Leu Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln
465 470 475 480
Lys Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr
485 490 495
Ile Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg
500 505 510
Glu Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro
515 520 525
Pro Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala
530 535 540
His Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln
545 550 555 560
Ala Asn Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe
565 570 575
Leu Arg Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met
580 585 590
Pro Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu
595 600 605
Lys Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly
610 615 620
Ala Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu
625 630 635 640
Ser Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln
645 650 655
Cys Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met
660 665 670
Ser Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys
675 680 685
Asp Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile
690 695 700
Lys Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu
705 710 715 720
Asp Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala
725 730 735
Leu Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln
740 745 750
Gly Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu
755 760 765
Thr Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg
770 775 780
Met Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly
785 790 795 800
Pro Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp
805 810 815
Val Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile
820 825 830
Ala Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu
835 840 845
Leu Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser
850 855 860
Pro Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met
865 870 875 880
Asn Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg
885 890 895
Pro Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg
900 905 910
Ala Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg
915 920 925
Glu Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu
930 935 940
Glu Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu
945 950 955 960
Asp Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr
965 970 975
Arg Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe
980 985 990
Glu Glu Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala
995 1000 1005
Glu Lys Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg
1010 1015 1020
Thr Ile Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr
1025 1030 1035
Leu Val Ser Gly Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro
1040 1045 1050
Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu
1055 1060 1065
Leu Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln
1070 1075 1080
His Glu Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu
1085 1090 1095
Ala Ser Leu Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly
1100 1105 1110
Pro Gly Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser
1115 1120 1125
Gln Leu Leu Glu Thr Leu Asn Gln Leu Ser Thr His Thr His Val
1130 1135 1140
Val Asp Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala
1145 1150 1155
Gln Leu Met Glu Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr
1160 1165 1170
Val Glu Lys Leu Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln
1175 1180 1185
Arg Pro Gly Ala Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser
1190 1195 1200
Ser Ala Phe Leu Arg Ala Lys Glu Glu Gln Gln Asp Asp Thr Val
1205 1210 1215
Tyr Met Gly Lys Val Thr Phe Ser Cys Ala Ala Gly Phe Gly Gln
1220 1225 1230
Arg His Arg Leu Val Leu Thr Gln Glu Gln Leu His Gln Leu His
1235 1240 1245
Ser Arg Leu Ile Ser
1250
<210> 28
<211> 1139
<212> PRT
<213> 智人
<400> 28
Met Met Arg Gln Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro
1 5 10 15
Lys Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser
20 25 30
Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu
35 40 45
Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr
50 55 60
Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser
65 70 75 80
Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys
85 90 95
Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys
100 105 110
Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys
115 120 125
Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu
130 135 140
Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met
145 150 155 160
Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp
165 170 175
Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu
180 185 190
Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu
195 200 205
Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr
210 215 220
Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu
225 230 235 240
Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys
245 250 255
Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg
260 265 270
Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr
275 280 285
Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu
290 295 300
Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val
305 310 315 320
Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met
325 330 335
Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg
340 345 350
Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg
355 360 365
Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys
370 375 380
Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu
385 390 395 400
Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro
405 410 415
Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala
420 425 430
Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn
435 440 445
Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg
450 455 460
Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg
465 470 475 480
Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe
485 490 495
Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala
500 505 510
Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu
515 520 525
Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser
530 535 540
Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala
545 550 555 560
His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln
565 570 575
Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr
580 585 590
Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys
595 600 605
Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp
610 615 620
Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly
625 630 635 640
Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser
645 650 655
Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro
660 665 670
Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln
675 680 685
Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val
690 695 700
Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro
705 710 715 720
Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala
725 730 735
Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr
740 745 750
Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys
755 760 765
Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro
770 775 780
Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu
785 790 795 800
Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr
805 810 815
Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu
820 825 830
Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser
835 840 845
Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu
850 855 860
Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu
865 870 875 880
Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
885 890 895
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile Glu
900 905 910
Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val Ser Gly
915 920 925
Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser Val Pro Gly
930 935 940
Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln Gln Ile Ser Ala
945 950 955 960
Met Arg Leu His Ile Ser Gln Leu Gln His Glu Asn Ser Ile Leu Lys
965 970 975
Gly Ala Gln Met Lys Ala Ser Leu Ala Ser Leu Pro Pro Leu His Val
980 985 990
Ala Lys Leu Ser His Glu Gly Pro Gly Ser Glu Leu Pro Ala Gly Ala
995 1000 1005
Leu Tyr Arg Lys Thr Ser Gln Leu Leu Glu Thr Leu Asn Gln Leu
1010 1015 1020
Ser Thr His Thr His Val Val Asp Ile Thr Arg Thr Ser Pro Ala
1025 1030 1035
Ala Lys Ser Pro Ser Ala Gln Leu Met Glu Gln Val Ala Gln Leu
1040 1045 1050
Lys Ser Leu Ser Asp Thr Val Glu Lys Leu Lys Asp Glu Val Leu
1055 1060 1065
Lys Glu Thr Val Ser Gln Arg Pro Gly Ala Thr Val Pro Thr Asp
1070 1075 1080
Phe Ala Thr Phe Pro Ser Ser Ala Phe Leu Arg Ala Lys Glu Glu
1085 1090 1095
Gln Gln Asp Asp Thr Val Tyr Met Gly Lys Val Thr Phe Ser Cys
1100 1105 1110
Ala Ala Gly Phe Gly Gln Arg His Arg Leu Val Leu Thr Gln Glu
1115 1120 1125
Gln Leu His Gln Leu His Ser Arg Leu Ile Ser
1130 1135
<210> 29
<211> 1236
<212> PRT
<213> 智人
<400> 29
Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg Val
1 5 10 15
Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly Ala
20 25 30
Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu Ala
35 40 45
Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr Cys
50 55 60
Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val Phe
65 70 75 80
Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser Ala
85 90 95
Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr Ser
100 105 110
Lys Leu Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser
115 120 125
Ser Leu Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser
130 135 140
Glu Pro Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro
145 150 155 160
Thr Pro Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro
165 170 175
Ser Lys Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu
180 185 190
Lys Leu Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu
195 200 205
Lys Glu Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp
210 215 220
Lys Ser Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys
225 230 235 240
Glu Ala Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr
245 250 255
Met Glu Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu
260 265 270
Asp Lys Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val
275 280 285
Glu Ala Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile
290 295 300
Leu Lys Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser
305 310 315 320
Tyr Gln Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala
325 330 335
Leu Val Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val
340 345 350
Lys Leu Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val
355 360 365
Arg Gln Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser
370 375 380
Thr Ile Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu
385 390 395 400
Glu Met Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys
405 410 415
Val Arg Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu
420 425 430
Met Asn Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu
435 440 445
Arg Glu Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys
450 455 460
Arg Val Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile
465 470 475 480
Lys Lys Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu
485 490 495
Leu Thr Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro
500 505 510
Pro Glu Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His
515 520 525
Ala Lys Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala
530 535 540
Asn Arg His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu
545 550 555 560
Arg Pro Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro
565 570 575
Arg Leu Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys
580 585 590
Phe Glu Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala
595 600 605
Ala Gly Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser
610 615 620
Leu Leu Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys
625 630 635 640
Ser Val Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser
645 650 655
Ala His Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp
660 665 670
Gln Leu Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys
675 680 685
Tyr Tyr Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp
690 695 700
Cys Thr Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu
705 710 715 720
Asp Cys Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly
725 730 735
Gly Gln Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr
740 745 750
Ser Cys Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met
755 760 765
Pro Gly Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro
770 775 780
Gln Val Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val
785 790 795 800
Val Ala Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala
805 810 815
Pro Leu Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu
820 825 830
Ala Phe Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro
835 840 845
Tyr Glu Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn
850 855 860
Lys Leu Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro
865 870 875 880
Pro Ser Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala
885 890 895
Glu Ile Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu
900 905 910
Thr Val Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu
915 920 925
Leu Ser Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp
930 935 940
Ser Ala Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg
945 950 955 960
Leu Glu Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu
965 970 975
Glu Thr Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu
980 985 990
Lys Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile
995 1000 1005
Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val
1010 1015 1020
Ser Gly Ile Ala Gly Gly Ala Ile Pro Gly Gln Ala Pro Gly Ser
1025 1030 1035
Val Pro Gly Pro Gly Leu Val Lys Asp Ser Pro Leu Leu Leu Gln
1040 1045 1050
Gln Ile Ser Ala Met Arg Leu His Ile Ser Gln Leu Gln His Glu
1055 1060 1065
Asn Ser Ile Leu Lys Gly Ala Gln Met Lys Ala Ser Leu Ala Ser
1070 1075 1080
Leu Pro Pro Leu His Val Ala Lys Leu Ser His Glu Gly Pro Gly
1085 1090 1095
Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg Lys Thr Ser Gln Leu
1100 1105 1110
Leu Glu Thr Leu Asn Gln Leu Ser Thr His Thr His Val Val Asp
1115 1120 1125
Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser Pro Ser Ala Gln Leu
1130 1135 1140
Met Glu Gln Val Ala Gln Leu Lys Ser Leu Ser Asp Thr Val Glu
1145 1150 1155
Lys Leu Lys Asp Glu Val Leu Lys Glu Thr Val Ser Gln Arg Pro
1160 1165 1170
Gly Ala Thr Val Pro Thr Asp Phe Ala Thr Phe Pro Ser Ser Ala
1175 1180 1185
Phe Leu Arg Ala Lys Glu Glu Gln Gln Asp Asp Thr Val Tyr Met
1190 1195 1200
Gly Lys Val Thr Phe Ser Cys Ala Ala Gly Phe Gly Gln Arg His
1205 1210 1215
Arg Leu Val Leu Thr Gln Glu Gln Leu His Gln Leu His Ser Arg
1220 1225 1230
Leu Ile Ser
1235
<210> 30
<211> 1271
<212> PRT
<213> 智人
<400> 30
Met Ala Gln Ser Lys Arg His Val Tyr Ser Arg Thr Pro Ser Gly Ser
1 5 10 15
Arg Met Ser Ala Glu Ala Ser Ala Arg Pro Leu Arg Val Gly Ser Arg
20 25 30
Val Glu Val Ile Gly Lys Gly His Arg Gly Thr Val Ala Tyr Val Gly
35 40 45
Ala Thr Leu Phe Ala Thr Gly Lys Trp Val Gly Val Ile Leu Asp Glu
50 55 60
Ala Lys Gly Lys Asn Asp Gly Thr Val Gln Gly Arg Lys Tyr Phe Thr
65 70 75 80
Cys Asp Glu Gly His Gly Ile Phe Val Arg Gln Ser Gln Ile Gln Val
85 90 95
Phe Glu Asp Gly Ala Asp Thr Thr Ser Pro Glu Thr Pro Asp Ser Ser
100 105 110
Ala Ser Lys Val Leu Lys Arg Glu Gly Thr Asp Thr Thr Ala Lys Thr
115 120 125
Ser Lys Leu Ala Pro Thr Ala Arg Lys Thr Thr Thr Arg Arg Pro Lys
130 135 140
Pro Thr Arg Pro Ala Ser Thr Gly Val Ala Gly Ala Ser Ser Ser Leu
145 150 155 160
Gly Pro Ser Gly Ser Ala Ser Ala Gly Glu Leu Ser Ser Ser Glu Pro
165 170 175
Ser Thr Pro Ala Gln Thr Pro Leu Ala Ala Pro Ile Ile Pro Thr Pro
180 185 190
Val Leu Thr Ser Pro Gly Ala Val Pro Pro Leu Pro Ser Pro Ser Lys
195 200 205
Glu Glu Glu Gly Leu Arg Ala Gln Val Arg Asp Leu Glu Glu Lys Leu
210 215 220
Glu Thr Leu Arg Leu Lys Arg Ala Glu Asp Lys Ala Lys Leu Lys Glu
225 230 235 240
Leu Glu Lys His Lys Ile Gln Leu Glu Gln Val Gln Glu Trp Lys Ser
245 250 255
Lys Met Gln Glu Gln Gln Ala Asp Leu Gln Arg Arg Leu Lys Glu Ala
260 265 270
Arg Lys Glu Ala Lys Glu Ala Leu Glu Ala Lys Glu Arg Tyr Met Glu
275 280 285
Glu Met Ala Asp Thr Ala Asp Ala Ile Glu Met Ala Thr Leu Asp Lys
290 295 300
Glu Met Ala Glu Glu Arg Ala Glu Ser Leu Gln Gln Glu Val Glu Ala
305 310 315 320
Leu Lys Glu Arg Val Asp Glu Leu Thr Thr Asp Leu Glu Ile Leu Lys
325 330 335
Ala Glu Ile Glu Glu Lys Gly Ser Asp Gly Ala Ala Ser Ser Tyr Gln
340 345 350
Leu Lys Gln Leu Glu Glu Gln Asn Ala Arg Leu Lys Asp Ala Leu Val
355 360 365
Arg Met Arg Asp Leu Ser Ser Ser Glu Lys Gln Glu His Val Lys Leu
370 375 380
Gln Lys Leu Met Glu Lys Lys Asn Gln Glu Leu Glu Val Val Arg Gln
385 390 395 400
Gln Arg Glu Arg Leu Gln Glu Glu Leu Ser Gln Ala Glu Ser Thr Ile
405 410 415
Asp Glu Leu Lys Glu Gln Val Asp Ala Ala Leu Gly Ala Glu Glu Met
420 425 430
Val Glu Met Leu Thr Asp Arg Asn Leu Asn Leu Glu Glu Lys Val Arg
435 440 445
Glu Leu Arg Glu Thr Val Gly Asp Leu Glu Ala Met Asn Glu Met Asn
450 455 460
Asp Glu Leu Gln Glu Asn Ala Arg Glu Thr Glu Leu Glu Leu Arg Glu
465 470 475 480
Gln Leu Asp Met Ala Gly Ala Arg Val Arg Glu Ala Gln Lys Arg Val
485 490 495
Glu Ala Ala Gln Glu Thr Val Ala Asp Tyr Gln Gln Thr Ile Lys Lys
500 505 510
Tyr Arg Gln Leu Thr Ala His Leu Gln Asp Val Asn Arg Glu Leu Thr
515 520 525
Asn Gln Gln Glu Ala Ser Val Glu Arg Gln Gln Gln Pro Pro Pro Glu
530 535 540
Thr Phe Asp Phe Lys Ile Lys Phe Ala Glu Thr Lys Ala His Ala Lys
545 550 555 560
Ala Ile Glu Met Glu Leu Arg Gln Met Glu Val Ala Gln Ala Asn Arg
565 570 575
His Met Ser Leu Leu Thr Ala Phe Met Pro Asp Ser Phe Leu Arg Pro
580 585 590
Gly Gly Asp His Asp Cys Val Leu Val Leu Leu Leu Met Pro Arg Leu
595 600 605
Ile Cys Lys Ala Glu Leu Ile Arg Lys Gln Ala Gln Glu Lys Phe Glu
610 615 620
Leu Ser Glu Asn Cys Ser Glu Arg Pro Gly Leu Arg Gly Ala Ala Gly
625 630 635 640
Glu Gln Leu Ser Phe Ala Ala Gly Leu Val Tyr Ser Leu Ser Leu Leu
645 650 655
Gln Ala Thr Leu His Arg Tyr Glu His Ala Leu Ser Gln Cys Ser Val
660 665 670
Asp Val Tyr Lys Lys Val Gly Ser Leu Tyr Pro Glu Met Ser Ala His
675 680 685
Glu Arg Ser Leu Asp Phe Leu Ile Glu Leu Leu His Lys Asp Gln Leu
690 695 700
Asp Glu Thr Val Asn Val Glu Pro Leu Thr Lys Ala Ile Lys Tyr Tyr
705 710 715 720
Gln His Leu Tyr Ser Ile His Leu Ala Glu Gln Pro Glu Asp Cys Thr
725 730 735
Met Gln Leu Ala Asp His Ile Lys Phe Thr Gln Ser Ala Leu Asp Cys
740 745 750
Met Ser Val Glu Val Gly Arg Leu Arg Ala Phe Leu Gln Gly Gly Gln
755 760 765
Glu Ala Thr Asp Ile Ala Leu Leu Leu Arg Asp Leu Glu Thr Ser Cys
770 775 780
Ser Asp Ile Arg Gln Phe Cys Lys Lys Ile Arg Arg Arg Met Pro Gly
785 790 795 800
Thr Asp Ala Pro Gly Ile Pro Ala Ala Leu Ala Phe Gly Pro Gln Val
805 810 815
Ser Asp Thr Leu Leu Asp Cys Arg Lys His Leu Thr Trp Val Val Ala
820 825 830
Val Leu Gln Glu Val Ala Ala Ala Ala Ala Gln Leu Ile Ala Pro Leu
835 840 845
Ala Glu Asn Glu Gly Leu Leu Val Ala Ala Leu Glu Glu Leu Ala Phe
850 855 860
Lys Ala Ser Glu Gln Ile Tyr Gly Thr Pro Ser Ser Ser Pro Tyr Glu
865 870 875 880
Cys Leu Arg Gln Ser Cys Asn Ile Leu Ile Ser Thr Met Asn Lys Leu
885 890 895
Ala Thr Ala Met Gln Glu Gly Glu Tyr Asp Ala Glu Arg Pro Pro Ser
900 905 910
Lys Pro Pro Pro Val Glu Leu Arg Ala Ala Ala Leu Arg Ala Glu Ile
915 920 925
Thr Asp Ala Glu Gly Leu Gly Leu Lys Leu Glu Asp Arg Glu Thr Val
930 935 940
Ile Lys Glu Leu Lys Lys Ser Leu Lys Ile Lys Gly Glu Glu Leu Ser
945 950 955 960
Glu Ala Asn Val Arg Leu Ser Leu Leu Glu Lys Lys Leu Asp Ser Ala
965 970 975
Ala Lys Asp Ala Asp Glu Arg Ile Glu Lys Val Gln Thr Arg Leu Glu
980 985 990
Glu Thr Gln Ala Leu Leu Arg Lys Lys Glu Lys Glu Phe Glu Glu Thr
995 1000 1005
Met Asp Ala Leu Gln Ala Asp Ile Asp Gln Leu Glu Ala Glu Lys
1010 1015 1020
Ala Glu Leu Lys Gln Arg Leu Asn Ser Gln Ser Lys Arg Thr Ile
1025 1030 1035
Glu Gly Leu Arg Gly Pro Pro Pro Ser Gly Ile Ala Thr Leu Val
1040 1045 1050
Ser Gly Ile Ala Gly Glu Glu Gln Gln Arg Gly Ala Ile Pro Gly
1055 1060 1065
Gln Ala Pro Gly Ser Val Pro Gly Pro Gly Leu Val Lys Asp Ser
1070 1075 1080
Pro Leu Leu Leu Gln Gln Ile Ser Ala Met Arg Leu His Ile Ser
1085 1090 1095
Gln Leu Gln His Glu Asn Ser Ile Leu Lys Gly Ala Gln Met Lys
1100 1105 1110
Ala Ser Leu Ala Ser Leu Pro Pro Leu His Val Ala Lys Leu Ser
1115 1120 1125
His Glu Gly Pro Gly Ser Glu Leu Pro Ala Gly Ala Leu Tyr Arg
1130 1135 1140
Lys Thr Ser Gln Leu Leu Glu Thr Leu Asn Gln Leu Ser Thr His
1145 1150 1155
Thr His Val Val Asp Ile Thr Arg Thr Ser Pro Ala Ala Lys Ser
1160 1165 1170
Pro Ser Ala Gln Leu Met Glu Gln Val Ala Gln Leu Lys Ser Leu
1175 1180 1185
Ser Asp Thr Val Glu Lys Leu Lys Asp Glu Val Leu Lys Glu Thr
1190 1195 1200
Val Ser Gln Arg Pro Gly Ala Thr Val Pro Thr Asp Phe Ala Thr
1205 1210 1215
Phe Pro Ser Ser Ala Phe Leu Arg Ala Lys Glu Glu Gln Gln Asp
1220 1225 1230
Asp Thr Val Tyr Met Gly Lys Val Thr Phe Ser Cys Ala Ala Gly
1235 1240 1245
Phe Gly Gln Arg His Arg Leu Val Leu Thr Gln Glu Gln Leu His
1250 1255 1260
Gln Leu His Ser Arg Leu Ile Ser
1265 1270
<210> 31
<211> 1114
<212> PRT
<213> 智人
<400> 31
Met Ala Lys Ala Thr Ser Gly Ala Ala Gly Leu Arg Leu Leu Leu Leu
1 5 10 15
Leu Leu Leu Pro Leu Leu Gly Lys Val Ala Leu Gly Leu Tyr Phe Ser
20 25 30
Arg Asp Ala Tyr Trp Glu Lys Leu Tyr Val Asp Gln Ala Ala Gly Thr
35 40 45
Pro Leu Leu Tyr Val His Ala Leu Arg Asp Ala Pro Glu Glu Val Pro
50 55 60
Ser Phe Arg Leu Gly Gln His Leu Tyr Gly Thr Tyr Arg Thr Arg Leu
65 70 75 80
His Glu Asn Asn Trp Ile Cys Ile Gln Glu Asp Thr Gly Leu Leu Tyr
85 90 95
Leu Asn Arg Ser Leu Asp His Ser Ser Trp Glu Lys Leu Ser Val Arg
100 105 110
Asn Arg Gly Phe Pro Leu Leu Thr Val Tyr Leu Lys Val Phe Leu Ser
115 120 125
Pro Thr Ser Leu Arg Glu Gly Glu Cys Gln Trp Pro Gly Cys Ala Arg
130 135 140
Val Tyr Phe Ser Phe Phe Asn Thr Ser Phe Pro Ala Cys Ser Ser Leu
145 150 155 160
Lys Pro Arg Glu Leu Cys Phe Pro Glu Thr Arg Pro Ser Phe Arg Ile
165 170 175
Arg Glu Asn Arg Pro Pro Gly Thr Phe His Gln Phe Arg Leu Leu Pro
180 185 190
Val Gln Phe Leu Cys Pro Asn Ile Ser Val Ala Tyr Arg Leu Leu Glu
195 200 205
Gly Glu Gly Leu Pro Phe Arg Cys Ala Pro Asp Ser Leu Glu Val Ser
210 215 220
Thr Arg Trp Ala Leu Asp Arg Glu Gln Arg Glu Lys Tyr Glu Leu Val
225 230 235 240
Ala Val Cys Thr Val His Ala Gly Ala Arg Glu Glu Val Val Met Val
245 250 255
Pro Phe Pro Val Thr Val Tyr Asp Glu Asp Asp Ser Ala Pro Thr Phe
260 265 270
Pro Ala Gly Val Asp Thr Ala Ser Ala Val Val Glu Phe Lys Arg Lys
275 280 285
Glu Asp Thr Val Val Ala Thr Leu Arg Val Phe Asp Ala Asp Val Val
290 295 300
Pro Ala Ser Gly Glu Leu Val Arg Arg Tyr Thr Ser Thr Leu Leu Pro
305 310 315 320
Gly Asp Thr Trp Ala Gln Gln Thr Phe Arg Val Glu His Trp Pro Asn
325 330 335
Glu Thr Ser Val Gln Ala Asn Gly Ser Phe Val Arg Ala Thr Val His
340 345 350
Asp Tyr Arg Leu Val Leu Asn Arg Asn Leu Ser Ile Ser Glu Asn Arg
355 360 365
Thr Met Gln Leu Ala Val Leu Val Asn Asp Ser Asp Phe Gln Gly Pro
370 375 380
Gly Ala Gly Val Leu Leu Leu His Phe Asn Val Ser Val Leu Pro Val
385 390 395 400
Ser Leu His Leu Pro Ser Thr Tyr Ser Leu Ser Val Ser Arg Arg Ala
405 410 415
Arg Arg Phe Ala Gln Ile Gly Lys Val Cys Val Glu Asn Cys Gln Ala
420 425 430
Phe Ser Gly Ile Asn Val Gln Tyr Lys Leu His Ser Ser Gly Ala Asn
435 440 445
Cys Ser Thr Leu Gly Val Val Thr Ser Ala Glu Asp Thr Ser Gly Ile
450 455 460
Leu Phe Val Asn Asp Thr Lys Ala Leu Arg Arg Pro Lys Cys Ala Glu
465 470 475 480
Leu His Tyr Met Val Val Ala Thr Asp Gln Gln Thr Ser Arg Gln Ala
485 490 495
Gln Ala Gln Leu Leu Val Thr Val Glu Gly Ser Tyr Val Ala Glu Glu
500 505 510
Ala Gly Cys Pro Leu Ser Cys Ala Val Ser Lys Arg Arg Leu Glu Cys
515 520 525
Glu Glu Cys Gly Gly Leu Gly Ser Pro Thr Gly Arg Cys Glu Trp Arg
530 535 540
Gln Gly Asp Gly Lys Gly Ile Thr Arg Asn Phe Ser Thr Cys Ser Pro
545 550 555 560
Ser Thr Lys Thr Cys Pro Asp Gly His Cys Asp Val Val Glu Thr Gln
565 570 575
Asp Ile Asn Ile Cys Pro Gln Asp Cys Leu Arg Gly Ser Ile Val Gly
580 585 590
Gly His Glu Pro Gly Glu Pro Arg Gly Ile Lys Ala Gly Tyr Gly Thr
595 600 605
Cys Asn Cys Phe Pro Glu Glu Glu Lys Cys Phe Cys Glu Pro Glu Asp
610 615 620
Ile Gln Asp Pro Leu Cys Asp Glu Leu Cys Arg Thr Val Ile Ala Ala
625 630 635 640
Ala Val Leu Phe Ser Phe Ile Val Ser Val Leu Leu Ser Ala Phe Cys
645 650 655
Ile His Cys Tyr His Lys Phe Ala His Lys Pro Pro Ile Ser Ser Ala
660 665 670
Glu Met Thr Phe Arg Arg Pro Ala Gln Ala Phe Pro Val Ser Tyr Ser
675 680 685
Ser Ser Gly Ala Arg Arg Pro Ser Leu Asp Ser Met Glu Asn Gln Val
690 695 700
Ser Val Asp Ala Phe Lys Ile Leu Glu Asp Pro Lys Trp Glu Phe Pro
705 710 715 720
Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly
725 730 735
Lys Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr
740 745 750
Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu
755 760 765
Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His
770 775 780
Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu
785 790 795 800
Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu
805 810 815
Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser
820 825 830
Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr Met
835 840 845
Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr
850 855 860
Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile
865 870 875 880
Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser
885 890 895
Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg
900 905 910
Ile Pro Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr
915 920 925
Thr Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile
930 935 940
Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu
945 950 955 960
Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys
965 970 975
Ser Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro
980 985 990
Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met
995 1000 1005
Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro
1010 1015 1020
Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr
1025 1030 1035
Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro
1040 1045 1050
Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Met Ser Asp Pro Asn
1055 1060 1065
Trp Pro Gly Glu Ser Pro Val Pro Leu Thr Arg Ala Asp Gly Thr
1070 1075 1080
Asn Thr Gly Phe Pro Arg Tyr Pro Asn Asp Ser Val Tyr Ala Asn
1085 1090 1095
Trp Met Leu Ser Pro Ser Ala Ala Lys Leu Met Asp Thr Phe Asp
1100 1105 1110
Ser
<210> 32
<211> 1072
<212> PRT
<213> 智人
<400> 32
Met Ala Lys Ala Thr Ser Gly Ala Ala Gly Leu Arg Leu Leu Leu Leu
1 5 10 15
Leu Leu Leu Pro Leu Leu Gly Lys Val Ala Leu Gly Leu Tyr Phe Ser
20 25 30
Arg Asp Ala Tyr Trp Glu Lys Leu Tyr Val Asp Gln Ala Ala Gly Thr
35 40 45
Pro Leu Leu Tyr Val His Ala Leu Arg Asp Ala Pro Glu Glu Val Pro
50 55 60
Ser Phe Arg Leu Gly Gln His Leu Tyr Gly Thr Tyr Arg Thr Arg Leu
65 70 75 80
His Glu Asn Asn Trp Ile Cys Ile Gln Glu Asp Thr Gly Leu Leu Tyr
85 90 95
Leu Asn Arg Ser Leu Asp His Ser Ser Trp Glu Lys Leu Ser Val Arg
100 105 110
Asn Arg Gly Phe Pro Leu Leu Thr Val Tyr Leu Lys Val Phe Leu Ser
115 120 125
Pro Thr Ser Leu Arg Glu Gly Glu Cys Gln Trp Pro Gly Cys Ala Arg
130 135 140
Val Tyr Phe Ser Phe Phe Asn Thr Ser Phe Pro Ala Cys Ser Ser Leu
145 150 155 160
Lys Pro Arg Glu Leu Cys Phe Pro Glu Thr Arg Pro Ser Phe Arg Ile
165 170 175
Arg Glu Asn Arg Pro Pro Gly Thr Phe His Gln Phe Arg Leu Leu Pro
180 185 190
Val Gln Phe Leu Cys Pro Asn Ile Ser Val Ala Tyr Arg Leu Leu Glu
195 200 205
Gly Glu Gly Leu Pro Phe Arg Cys Ala Pro Asp Ser Leu Glu Val Ser
210 215 220
Thr Arg Trp Ala Leu Asp Arg Glu Gln Arg Glu Lys Tyr Glu Leu Val
225 230 235 240
Ala Val Cys Thr Val His Ala Gly Ala Arg Glu Glu Val Val Met Val
245 250 255
Pro Phe Pro Val Thr Val Tyr Asp Glu Asp Asp Ser Ala Pro Thr Phe
260 265 270
Pro Ala Gly Val Asp Thr Ala Ser Ala Val Val Glu Phe Lys Arg Lys
275 280 285
Glu Asp Thr Val Val Ala Thr Leu Arg Val Phe Asp Ala Asp Val Val
290 295 300
Pro Ala Ser Gly Glu Leu Val Arg Arg Tyr Thr Ser Thr Leu Leu Pro
305 310 315 320
Gly Asp Thr Trp Ala Gln Gln Thr Phe Arg Val Glu His Trp Pro Asn
325 330 335
Glu Thr Ser Val Gln Ala Asn Gly Ser Phe Val Arg Ala Thr Val His
340 345 350
Asp Tyr Arg Leu Val Leu Asn Arg Asn Leu Ser Ile Ser Glu Asn Arg
355 360 365
Thr Met Gln Leu Ala Val Leu Val Asn Asp Ser Asp Phe Gln Gly Pro
370 375 380
Gly Ala Gly Val Leu Leu Leu His Phe Asn Val Ser Val Leu Pro Val
385 390 395 400
Ser Leu His Leu Pro Ser Thr Tyr Ser Leu Ser Val Ser Arg Arg Ala
405 410 415
Arg Arg Phe Ala Gln Ile Gly Lys Val Cys Val Glu Asn Cys Gln Ala
420 425 430
Phe Ser Gly Ile Asn Val Gln Tyr Lys Leu His Ser Ser Gly Ala Asn
435 440 445
Cys Ser Thr Leu Gly Val Val Thr Ser Ala Glu Asp Thr Ser Gly Ile
450 455 460
Leu Phe Val Asn Asp Thr Lys Ala Leu Arg Arg Pro Lys Cys Ala Glu
465 470 475 480
Leu His Tyr Met Val Val Ala Thr Asp Gln Gln Thr Ser Arg Gln Ala
485 490 495
Gln Ala Gln Leu Leu Val Thr Val Glu Gly Ser Tyr Val Ala Glu Glu
500 505 510
Ala Gly Cys Pro Leu Ser Cys Ala Val Ser Lys Arg Arg Leu Glu Cys
515 520 525
Glu Glu Cys Gly Gly Leu Gly Ser Pro Thr Gly Arg Cys Glu Trp Arg
530 535 540
Gln Gly Asp Gly Lys Gly Ile Thr Arg Asn Phe Ser Thr Cys Ser Pro
545 550 555 560
Ser Thr Lys Thr Cys Pro Asp Gly His Cys Asp Val Val Glu Thr Gln
565 570 575
Asp Ile Asn Ile Cys Pro Gln Asp Cys Leu Arg Gly Ser Ile Val Gly
580 585 590
Gly His Glu Pro Gly Glu Pro Arg Gly Ile Lys Ala Gly Tyr Gly Thr
595 600 605
Cys Asn Cys Phe Pro Glu Glu Glu Lys Cys Phe Cys Glu Pro Glu Asp
610 615 620
Ile Gln Asp Pro Leu Cys Asp Glu Leu Cys Arg Thr Val Ile Ala Ala
625 630 635 640
Ala Val Leu Phe Ser Phe Ile Val Ser Val Leu Leu Ser Ala Phe Cys
645 650 655
Ile His Cys Tyr His Lys Phe Ala His Lys Pro Pro Ile Ser Ser Ala
660 665 670
Glu Met Thr Phe Arg Arg Pro Ala Gln Ala Phe Pro Val Ser Tyr Ser
675 680 685
Ser Ser Gly Ala Arg Arg Pro Ser Leu Asp Ser Met Glu Asn Gln Val
690 695 700
Ser Val Asp Ala Phe Lys Ile Leu Glu Asp Pro Lys Trp Glu Phe Pro
705 710 715 720
Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly
725 730 735
Lys Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr
740 745 750
Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu
755 760 765
Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His
770 775 780
Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu
785 790 795 800
Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu
805 810 815
Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser
820 825 830
Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr Met
835 840 845
Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr
850 855 860
Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile
865 870 875 880
Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser
885 890 895
Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg
900 905 910
Ile Pro Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr
915 920 925
Thr Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile
930 935 940
Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu
945 950 955 960
Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys
965 970 975
Ser Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro
980 985 990
Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met
995 1000 1005
Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro
1010 1015 1020
Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr
1025 1030 1035
Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro
1040 1045 1050
Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Arg Ile Ser His Ala
1055 1060 1065
Phe Thr Arg Phe
1070
<210> 33
<211> 26
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 33
tgtccagctt tgtgcctgat tgatgt 26
<210> 34
<211> 26
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 34
gctgggcact gaagagaaag gaatgc 26
<210> 35
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 35
agcaggatga gtgcggaggc aagc 24
<210> 36
<211> 33
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 36
ttaactatca aacgtgtcca ttaattttgc cgc 33
<210> 37
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 37
agtactgggg tggctggg 18
<210> 38
<211> 19
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 38
cactttggac aaggagatg 19
<210> 39
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 39
acagaactgg agctgcgg 18
<210> 40
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 40
ggactggtgt actcgctg 18
<210> 41
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 41
tcctagactg caggaaacac 20
<210> 42
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 42
catcgagaaa gtccagac 18
<210> 43
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 43
gctgctggag acattgaa 18
<210> 44
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 44
tcactgctgc tcagctca 18
<210> 45
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 45
gaggatccaa agtgggaatt 20
<210> 46
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 46
agtatctggc cgagatgaag 20
<210> 47
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 47
gcaaagacct ggagaagatg 20
<210> 48
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 48
aggacgttga actctgacag 20
<210> 49
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 49
cctttgcttc atccagaatc 20
<210> 50
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 50
gattttgtgt ttctccagct ct 22
<210> 51
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 51
cctgcttctc tgaggaagaa 20
<210> 52
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 52
gggccttagt ctcagcaaac 20
<210> 53
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 53
gagcactctg cgtgaactta 20
<210> 54
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 54
cagcttgttc atggtactga t 21
<210> 55
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 55
tggtgagtcc ttcaccag 18
<210> 56
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 56
cctagagttt ttccaagaac ca 22
<210> 57
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 57
catttaactg gaatccgacc 20
<210> 58
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 58
gactctctcc aggccagttc 20
<210> 59
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 59
ggctatcaga agtaaaacca cc 22
<210> 60
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 60
cgagagctga tggcacta 18
<210> 61
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 61
cttcatcaca agtgaagtac ttcc 24
<210> 62
<211> 19
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 62
cgtactccac gatgaggag 19
<210> 63
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 63
gattctggat gaagcaaagg 20
<210> 64
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 64
ggaagtactt cacttgtgat gaag 24
<210> 65
<211> 18
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 65
cccagccacc ccagtact 18
<210> 66
<211> 17
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 66
gtaaaacgac ggccagt 17
<210> 67
<211> 17
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 67
gttttcccag tcacgac 17
<210> 68
<211> 17
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 68
caggaaacag ctatgac 17
<210> 69
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 69
ctggagccac agtacccact 20
<210> 70
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 70
tccaaattcg ccttctccta 20
<210> 71
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 71
ttcatcagcc ttcctcaggg aggat 25
<210> 72
<211> 41
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 72
ggggacaagt ttgtacaaaa aagcaggctt cgccaccagc a 41
<210> 73
<211> 42
<212> DNA
<213> 人工序列
<220>
<223> 引物
<400> 73
ggggaccact ttgtacaaga aagctgggtt ttaactatca aa 42
<210> 74
<211> 21
<212> RNA
<213> 人工序列
<220>
<223> RET siRNA1
<220>
<221> misc_feature
<223> 胸腺嘧啶n链
<220>
<221> misc_feature
<222> (20)..(21)
<223> 胸腺嘧啶n链
<400> 74
cacaugucau caaauuguan n 21
<210> 75
<211> 21
<212> RNA
<213> 人工序列
<220>
<223> RET siRNA2
<220>
<221> misc_feature
<222> (20)..(21)
<223> 胸腺嘧啶n链
<400> 75
ggauugaaaa caaacucuan n 21
<210> 76
<211> 21
<212> RNA
<213> 人工序列
<220>
<223> RET siRNA
<220>
<221> misc_feature
<222> (20)..(21)
<223> 胸腺嘧啶n链
<400> 76
gcuugucccg agauguuuan n 21
<210> 77
<211> 21
<212> RNA
<213> 人工序列
<220>
<223> RET siRNA3
<220>
<221> misc_feature
<222> (20)..(21)
<223> 胸腺嘧啶n链
<400> 77
ccacugcuac cacaaguuun n 21

Claims (9)

1.一种融合多肽，其具有激酶活性和/或细胞增殖作用，其中DCTN1蛋白质的N-端部分与RET蛋白质的C-端部分融合，所述融合多肽的氨基酸序列为SEQ ID NO：18所示的氨基酸序列。
2.一种多核苷酸，其编码根据权利要求1所述的多肽。
3.根据权利要求2所述的多核苷酸，其选自以下(g)和(h)：
(g)多核苷酸，其包括SEQ ID NO：17所示的碱基序列；
(h)多核苷酸，其在严格条件下与包括与SEQ ID NO：17所示的碱基序列互补的碱基序列的多核苷酸杂交。
4.一种表达载体，其包括根据权利要求2或3所述的多核苷酸。
5.一种用权利要求2或3所述的多核苷酸转染的细胞。
6.一种引物或探针，其用于检测样品中根据权利要求2或3所述的多核苷酸的存在，所述引物或探针是选自以下(k)和(l)的多核苷酸：
(k)多核苷酸，其是与编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的融合点杂交的探针；和
(1)多核苷酸，其是一组设计成将融合点夹在编码DCTN1蛋白质的多核苷酸和编码RET蛋白质的多核苷酸之间的有义引物和反义引物。
7.一种用于检测癌症的生物标志物，所述生物标志物包括选自根据权利要求1所述的多肽和编码所述多肽的多核苷酸的至少一种。
8.抑制RET的化合物在制造用于癌症治疗的药物组合物中的用途，所述药物组合物用于治疗对于DCTN1基因和RET基因的融合基因呈阳性、和/或对于DCTN1蛋白质和RET蛋白质的融合蛋白呈阳性的癌症患者，所述癌症为甲状腺癌，
其中所述化合物选自：
4-氨基-1-(叔丁基)-N-(5-甲基-1H-吡唑-3-基)-1H-吡唑并[3,4-d]嘧啶-3-甲酰胺；
4-氨基-7-(叔丁基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
4-氨基-7-(1-氟-2-甲基丙-2-基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
4-氨基-N-(5-甲基-1H-吡唑-3-基)-7-(1-甲基环丙基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
4-氨基-7-(2-环丙基丙-2-基)-N-(5-甲基-1H-吡唑-3-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-(3-吗啉代丙-1-炔-1-基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-((四氢-2H-吡喃-4-基)乙炔基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
(R)-4-氨基-N-[4-(甲氧基甲基)苯基]-7-(1-甲基环丙基)-6-((四氢呋喃-2-基)甲氧基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
4-氨基-N-[4-(甲氧基甲基)苯基]-6-(((1-甲基-1H-吡唑-4-基)乙炔基)-7-(1-甲基环丙基)-7H-吡咯并[2,3-d]嘧啶-5-甲酰胺；
卡博替尼；
凡德他尼；和
乐伐替尼。
9.根据权利要求6所述的引物或探针在制造检测试剂中的用途，所述检测试剂用于检测根据权利要求1所述的多肽的存在、或者根据权利要求2或3所述的多核苷酸的存在。
CN201880054583.2A 2017-08-21 2018-08-20 Dctn1蛋白质与ret蛋白质的融合蛋白 Active CN111032696B (zh)

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