CN112501204A - Il21r基因人源化非人动物及其构建方法和应用 - Google Patents
Il21r基因人源化非人动物及其构建方法和应用 Download PDFInfo
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- CN112501204A CN112501204A CN202110170978.2A CN202110170978A CN112501204A CN 112501204 A CN112501204 A CN 112501204A CN 202110170978 A CN202110170978 A CN 202110170978A CN 112501204 A CN112501204 A CN 112501204A
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Abstract
本发明提供了一种IL21R基因人源化改造的非人动物的构建方法、一种人源化IL21R蛋白、一种人源化IL21R基因、一种IL21R基因的靶向载体和其在生物医药领域的应用,利用同源重组的方式将部分编码人IL21R蛋白的核苷酸序列导入非人动物基因组中,该非人动物体内能正常表达人源化IL21R蛋白,可以用于人IL21R信号机理研究、炎症、肿瘤及自身免疫性疾病药物筛选,对免疫靶点的新药研发具有重要的应用价值。
Description
技术领域
本发明属于动物基因工程和基因遗传修饰领域,具体地说,涉及一种IL21R基因人源化的非人动物及其构建方法和在生物医药领域的应用。
背景技术
IL21R全名是interleukin 21 receptor。它是一种I型跨膜受体蛋白,主要表达在脾、胸腺、淋巴结及外周血液淋巴细胞。IL21R是由IL-21R亚单位和γc组成的复合物, 其中IL-21R亚单位为配体识别结合部位,γc为信号传导单位。IL-21R具有广泛的生物学功能,在结合其配体IL-21后,通过激活JAKs-STATs信号通路,刺激T细胞和NK细胞增殖、调节B细胞和树突状细胞存活和分化,从而参与机体的固有免疫和适应性免疫。作为新型的免疫调节因子,IL-21R及其配体参与了多种自身免疫性疾病的发生发展,通过调节IL-21R的表达水平或阻断其信号传导通路为自身免疫性疾病及其他相关疾病的治疗提供了新的研究方向。
随着基因工程技术的不断发展和成熟,用人类基因替代或置换动物的同源性基因已经实现,通过这种方式开发人源化实验动物模型是动物模型未来的发展方向。其中基因人源化动物模型,即,利用基因编辑技术,用人源正常或突变基因替换动物基因组的同源基因,可建立更接近人类生理或疾病特征的正常或突变基因动物模型。基因人源化动物不但本身具有重要应用价值,如通过基因人源化可改进和提升细胞或组织移植人源化动物模型,更重要的是,由于人类基因片段的插入,动物体内可表达或部分表达人源蛋白,可作为仅能识别人蛋白氨基酸序列的药物的靶点,为在动物水平进行抗人抗体及其它药物的筛选提供了可能。然而,由于动物与人类在生理学及病理学方面存在差异,加上基因(即遗传因子)的复杂性,如何能构建出“有效”的人源化动物模型用于新药研发仍是最大的挑战(Scheer N, Snaith M, Wolf CR, Seibler J. Generation and utility ofgenetically humanized mouse models, Drug Discov Today; 18(23-24):1200-11,2013)。
目前关于IL21R基因的动物模型主要为基因敲除鼠,而且主要用于肿瘤相关疾病机制研究。鉴于IL21R在肿瘤、自身免疫性疾病等多种疾病发生过程中的广泛参与性以及靶向该信号通路的巨大应用价值,为了使临床前期的试验更有效并使研发失败最小化,本领域仍急需开发人源化IL21R信号通路相关的非人动物模型。
发明内容
本发明的第一方面,提供了一种IL21R基因人源化改造的非人动物的构建方法,所述的非人动物的基因组中包括编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列,所述的构建方法包括用包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列替换至非人动物IL21R基因座。
优选的,所述的非人动物的基因组中包括SEQ ID NO:5所示核苷酸序列,所述的构建方法包括用包含SEQ ID NO:5所示核苷酸序列替换至非人动物IL21R基因座。
优选的,所述的插入或替换至非人动物IL21R基因座为插入或替换非人动物IL21R基因中编码SEQ ID NO:1第24至211位的核苷酸序列,或者插入或替换SEQ ID NO:24所示序列相同的核苷酸序列。
优选的,所述的构建方法包括插入、翻转、敲除或替换。
优选的,所述的非人动物的基因组中包含人IL21R核苷酸序列的3号外显子的部分、4号至5号外显子的全部和6号外显子的部分,进一步优选的,包含3-4号内含子和/或5-6号内含子,更优选的,包含3-6号外显子之间的任一内含子;其中,所述人IL21R核苷酸序列的3号外显子的部分至少包含从编码人IL21R蛋白胞外区N端1-5(例如1、2、3、4、5)个氨基酸的核苷酸序列开始至3号外显子最后1个核苷酸序列为止,6号外显子的部分至少包含从6号外显子第一个核苷酸开始至6号外显子编码的氨基酸C端1-20(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20)个氨基酸的核苷酸序列为止。
优选的,所述的构建方法包括用包含人IL21R核苷酸序列的3号至6号外显子的全部或部分核苷酸序列插入或替换到非人动物IL21R基因座上,进一步优选的,用包含人IL21R核苷酸序列的3号外显子的部分、4号至5号外显子的全部和6号外显子的部分核苷酸序列插入或替换到非人动物IL21R基因座上,更优选的,包含3-4号内含子和/或5-6号内含子,更进一步优选的,包含3-6号外显子之间的任一内含子;其中,所述人IL21R基因的3号外显子的部分至少包含从编码人IL21R蛋白胞外区N端1-5(例如1、2、3、4、5)个氨基酸的核苷酸序列开始至3号外显子最后1个核苷酸序列为止,6号外显子的部分至少包含从6号外显子第一个核苷酸开始至6号外显子编码的氨基酸C端1-20(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20)个氨基酸的核苷酸序列为止。
在本发明的一个具体实施方式中,所述的构建方法包括用包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列或者包含SEQ ID NO:5所示核苷酸序列替换至非人动物IL21R基因的相应区域。
优选的,所述的构建方法包括用包含人IL21R核苷酸序列的3号至6号外显子全部或部分替换非人动物IL21R核苷酸序列的3号至6号外显子的全部或部分;其中,所述非人动物IL21R核苷酸序列包含编码非人动物3号外显子的部分、4号至5号外显子的全部、6号外显子的部分核苷酸序列,优选的,包含3-4号内含子和/或5-6号内含子,进一步优选的,包含3-6号外显子之间的任一内含子,其中,所述非人动物IL21R核苷酸序列的3号外显子的部分至少包含从编码IL21R蛋白胞外区N端1-5(例如1、2、3、4、5)个氨基酸的核苷酸序列开始至3号外显子最后1个核苷酸为止,6号外显子的部分至少包含从6号外显子第一个核苷酸开始至6号外显子编码的氨基酸C端1-20(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20)个氨基酸的核苷酸序列为止。
优选的,所述的构建方法包括用包含所述人源化IL21R基因的核苷酸序列插入或替换到非人动物IL21R基因座上。
优选的,所述的构建方法包括用包含编码所述人源化IL21R蛋白的核苷酸序列插入或替换到非人动物IL21R基因座上。
优选的,所述的插入或替换位点为IL21R基因的内源调控元件之后。
优选的,所述的插入为首先破坏非人动物内源IL21R基因的编码框,随后进行插入操作,或者所述的插入步骤既可在内源IL21R基因处造成移码突变又可以实现插入人源序列的步骤。
优选的,所述的非人动物中人源化IL21R基因是纯合或杂合的。
优选的,所述非人动物的基因组中至少一个染色体上包含人源化IL21R基因。
优选的,所述的非人动物中至少一个细胞表达人或人源化IL21R蛋白。
优选的,使用基因编辑技术进行IL21R基因人源化改造的非人动物的构建,所述的基因编辑技术包括利用胚胎干细胞的基因打靶技术、CRISPR/Cas9技术、锌指核酸酶技术、转录激活子样效应因子核酸酶技术、归巢核酸内切酶或其他分子生物学技术。
本发明所述的非人动物为啮齿类动物;优选的,所述的啮齿类动物为大鼠或小鼠。
优选的,使用靶向载体进行IL21R基因人源化改造的非人动物的构建,所述靶向载体包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列或SEQ ID NO:5所示核苷酸序列,任选地,所述的靶向载体还包含与待改变的转换区5’端同源的DNA片段,即5’臂,和/或,所述的靶向载体还包含与待改变的转换区3’端同源的DNA片段,即3’臂,进一步优选的,所述的5’臂选自非人动物IL21R基因基因组DNA的100-10000个长度的核苷酸;优选的,所述的5’臂与NCBI登录号为NC_000073.7至少具有90%同源性的核苷酸;进一步优选的,所述5’臂序列与SEQ ID NO:3至少具有90%同源性,或者如SEQ ID NO:3所示;优选的,所述的3’臂选自非人动物IL21R基因基因组DNA的100-10000个长度的核苷酸;优选的,所述的3’臂与NCBI登录号为NC_000073.7至少具有90%同源性的核苷酸;进一步优选的,所述的3’臂序列与SEQID NO:4至少具有90%同源性,或者如SEQ ID NO:4所示。
优选的,所述的靶向载体包含人IL21R的3号至6号外显子的全部或部分核苷酸序列;进一步优选的,包含3号外显子的部分、4号至5号外显子的全部和6号外显子的部分,更优选的,包含3-4号内含子和/或5-6号内含子,更进一步优选的,包含3-6号外显子之间的任一内含子,其中,所述人IL21R的核苷酸序列的3号外显子的部分至少包含从编码人IL21R蛋白胞外区N端1-5(例如1、2、3、4、5)个氨基酸的核苷酸序列开始至3号外显子最后一个核苷酸为止,6号外显子的部分至少包含从6号外显子第一个核苷酸开始至6号外显子编码的氨基酸C端1-20(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20)个氨基酸的核苷酸序列为止。
优选的,所述的待改变的转换区位于非人动物IL21R基因座上。进一步优选的,位于非人动物IL21R基因的3号外显子至6号外显子上。
在本发明的一个具体实施方式中,所述的构建方法包括将上述靶向载体导入非人动物细胞中,培养该细胞(优选为胚胎干细胞),然后将培养后的细胞移植至雌性非人动物输卵管内,允许其发育,鉴定筛选获得非人动物。
优选的,所述的非人动物体内表达人或人源化IL21R蛋白,所述的人源化IL21R蛋白包含人IL21R蛋白的胞外区,任选地,包含与SEQ ID NO:2第24至211位或SEQ ID NO:23具有至少70%、80%、85%、90%、95%或至少99%同一性的氨基酸序列或者包含与SEQ ID NO:2第24至211位或SEQ ID NO:23所示氨基酸序列一致的氨基酸序列,同时内源IL21R蛋白的表达降低或缺失。
优选的,所述的人源化IL21R蛋白包含人IL21R蛋白的胞外区的全部或部分,进一步优选的,包含胞外区的部分,更优选的,所述的胞外区的部分包含N端去除0-5(例如1、2、3、4、5)个氨基酸和/或C端去除0-25(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25)个氨基酸的人IL21R蛋白胞外区。
优选的,所述的人源化IL21R蛋白还包含非人动物IL21R蛋白的部分,优选为非人动物IL21R蛋白的信号肽、胞外区、跨膜区和/或胞质区。
在本发明的一个具体实施方式中,所述的人源化IL21R蛋白包含下列组中的一种:
a)SEQ ID NO:23或SEQ ID NO:2第24至211位所示氨基酸序列的部分或全部;
b)与SEQ ID NO:23或SEQ ID NO:2第24至211位所示氨基酸的序列同一性程度为至少大约为90%、91%、92%、93%、94%、95%、96%、97%、98%或至少99%;
c)与SEQ ID NO:23或SEQ ID NO:2第24至211位所示的氨基酸的序列差异不超过10、9、8、7、6、5、4、3、2或不超过1个氨基酸;
d)具有SEQ ID NO:23或SEQ ID NO:2第24至211位所示的,包括取代、缺失和/或插入一个或多个氨基酸残基的氨基酸序列。
优选的,所述的非人动物的基因组中包含人源化IL21R基因,所述的人源化IL21R基因编码人源化IL21R蛋白。
优选的,所述的人源化IL21R基因包含SEQ ID NO:5所示的核苷酸序列,进一步优选的,所述的非人动物中包含的IL21R基因转录的mRNA序列包含SEQ ID NO:8所示的核苷酸序列。
在本发明的一个具体实施方式中,所述的人源化IL21R基因包含下列组中的一种:
a)人源化IL21R基因的mRNA序列为SEQ ID NO:8所示的序列的部分或全部;
b)人源化IL21R基因的mRNA序列与SEQ ID NO:8所示的核苷酸序列的部分或全部的同一性程度为至少大约为90%、91%、92%、93%、94%、95%、96%、97%、98%或至少99%;
c)人源化IL21R基因的mRNA序列与SEQ ID NO:8所示的核苷酸序列差异不超过10、9、8、7、6、5、4、3、2或不超过1个核苷酸;
d)人源化IL21R基因的mRNA序列具有SEQ ID NO:8所示的核苷酸序列所示的,包括取代、缺失和/或插入一个或多个核苷酸的核苷酸序列。
本发明的第二方面,提供了一种IL21R基因人源化改造的非人动物,所述的非人动物采用上述构建方法获得。
本发明的第三方面,提供了一种靶向载体,所述的靶向载体包含人IL21R核苷酸序列的部分,优选的,所述的靶向载体包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列或者SEQ ID NO:5所示核苷酸序列,任选地,所述的靶向载体还包含与待改变的转换区5’端同源的DNA片段,即5’臂,和/或,所述的靶向载体还包含与待改变的转换区3’端同源的DNA片段,即3’臂,优选的,所述的5’臂选自非人动物IL21R基因基因组DNA的100-10000个长度的核苷酸;优选的,所述的5’臂与NCBI登录号为NC_000073.7至少具有90%同源性的核苷酸;进一步优选的,所述5’臂序列与SEQ ID NO:3至少具有90%同源性,或者如SEQ ID NO:3所示;优选的,所述的3’臂选自非人动物IL21R基因基因组DNA的100-10000个长度的核苷酸;优选的,所述的3’臂与NCBI登录号为NC_000073.7至少具有90%同源性的核苷酸;进一步优选的,所述的3’臂序列与SEQ ID NO:4至少具有90%同源性,或者如SEQ ID NO:4所示。
优选的,所述的人IL21R核苷酸序列的部分包含人IL21R的3号至6号外显子的全部或部分核苷酸序列;进一步优选的,包含3号外显子的部分、4号至5号外显子的全部和6号外显子的部分,更优选的,包含3-4号内含子和/或5-6号内含子,更进一步优选的,包含3-6号外显子之间的任一内含子,其中,所述人IL21R的核苷酸序列的3号外显子的部分至少包含从编码人IL21R蛋白胞外区N端1-5(例如1、2、3、4、5)个氨基酸的核苷酸序列开始至3号外显子最后一个核苷酸为止,6号外显子的部分至少包含从6号外显子第一个核苷酸开始至6号外显子编码的氨基酸C端1-20(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20)个氨基酸的核苷酸序列为止。
优选的,所述的待改变的转换区位于非人动物IL21R基因座上,进一步优选的,所述的待改变的转换区位于非人动物IL21R基因3号至6号外显子上。
本发明所述的非人动物为啮齿类动物;优选的,所述的啮齿类动物为大鼠或小鼠。
优选的,所述的靶向载体还包含标记基因,进一步优选的,所述标记基因为负筛选标记的编码基因,更进一步优选的,所述负筛选标记的编码基因为白喉毒素A亚基的编码基因(DTA)。
在本发明的一个具体实施方式中,所述的靶向载体中还包括阳性克隆筛选的抗性基因,进一步优选的,所述阳性克隆筛选的抗性基因为新霉素磷酸转移酶编码序列Neo。
在本发明的一个具体实施方式中,所述的靶向载体中还包括特异性重组系统,进一步优选的,所述特异性重组系统为Frt重组位点(也可选择常规的LoxP重组系统),所述的特异性重组系统为具有两个Frt重组位点,分别连接在抗性基因的两侧。
本发明的第四方面,提供了一种包含上述靶向载体的细胞。
本发明的第五方面,提供了上述靶向载体,或者上述的细胞在IL21R基因修饰中的应用,优选的,所述的应用包括但不限于翻转、敲除、插入或替换。
本发明的第六方面,涉及一种IL21R基因改造的人源化细胞,所述的人源化IL21R基因改造细胞的基因组中包括人IL21R基因的3号外显子至6号外显子。优选的,所述的人IL21R基因编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列或包含SEQ ID NO:5所示核苷酸序列,其通过内源性IL21R调控元件调控;该人源化IL21R基因改造细胞体内表达人或人源化IL21R蛋白,同时内源IL21R蛋白的表达降低或缺失。优选的,所述的人IL21R基因通过内源性IL21R调控元件调控。
本发明的第七方面,涉及一种IL21R基因缺失的细胞,所述的IL21R基因缺失的细胞缺失内源IL21R基因的3号外显子至6号外显子。
本发明的第八方面,涉及一种荷瘤动物模型的制备方法,所述的动物模型的制备方法包括通过上述的人源化IL21R基因改造非人动物制备荷瘤动物模型的步骤。
优选的,所述的荷瘤动物模型的制备方法还包括在上述人源化基因改造非人动物或其后代植入肿瘤细胞的步骤。
本发明的第九方面,提供了一种上述的制备方法获得的荷瘤动物模型。
本发明的第十方面,涉及一种细胞或细胞系或原代细胞培养物,所述细胞或细胞系或原代细胞培养物来源于上述的非人动物或上述的荷瘤动物模型。
本发明的第十一方面,涉及一种组织或器官或其培养物,所述组织或器官或其培养物来源于上述的非人动物或上述的荷瘤动物模型。
优选的,所述的组织或器官或其培养物为脾脏、肿瘤或其培养物。
本发明的第十二方面,提供了一种人源化IL21R蛋白,所述的人源化IL21R蛋白包含人IL21R蛋白的全部或部分,所述的人源化IL21R蛋白包含人IL21R蛋白的胞外区,任选地,包含与SEQ ID NO:2第24至211位或SEQ ID NO:23具有至少70%、80%、85%、90%、95%或至少99%同一性的氨基酸序列或者包含与SEQ ID NO:2第24至211位或SEQ ID NO:23所示氨基酸序列一致的氨基酸序列。
优选的,所述的人源化IL21R蛋白包含人IL21R蛋白的胞外区的全部或部分,进一步优选的,包含胞外区的部分,更优选的,所述的胞外区的部分包含N端去除0-5(例如1、2、3、4、5)个氨基酸和/或C端去除0-25(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25)个氨基酸的人IL21R蛋白胞外区。
优选的,所述的人源化IL21R蛋白还包含非人动物IL21R蛋白的部分,优选为非人动物IL21R蛋白的信号肽、胞外区、跨膜区、胞质区。
优选的,所述的人源化IL21R蛋白包含人IL21R基因的3号外显子至6号外显子编码的氨基酸序列,和非人动物IL21R蛋白的氨基酸序列。
在本发明的一个具体实施方式中,所述的人源化IL21R蛋白包含下列组中的一种:
a)SEQ ID NO:23或SEQ ID NO:2第24至211位所示氨基酸序列的部分或全部;
b)与SEQ ID NO:23或SEQ ID NO:2第24至211位所示氨基酸的序列同一性程度为至少大约为90%、91%、92%、93%、94%、95%、96%、97%、98%或至少99%;
c)与SEQ ID NO:23或SEQ ID NO:2第24至211位所示的氨基酸的序列差异不超过10、9、8、7、6、5、4、3、2或不超过1个氨基酸;
d)具有SEQ ID NO:23或SEQ ID NO:2第24至211位所示的,包括取代、缺失和/或插入一个或多个氨基酸残基的氨基酸序列。
本发明的第十三方面,提供了一种编码上述人源化IL21R蛋白的人源化IL21R基因,所述的人源化IL21R基因包含人IL21R基因的3号外显子至6号外显子,和非人动物IL21R基因的核苷酸序列。
优选的,所述的人源化IL21R基因包含SEQ ID NO:5所示的核苷酸序列。
优选的,所述的人源化IL21R基因转录的mRNA序列包含SEQ ID NO:8所示的核苷酸序列。
在本发明的一个具体实施方式中,所述的人源化IL21R基因中包含的人IL21R核苷酸序列的部分选自下列组中的一种:
(A)包含SEQ ID NO:5所示核苷酸序列的全部或部分;
(B)包含与SEQ ID NO:5所示核苷酸序列的同一性至少为75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%或至少99%的核苷酸序列;
(C)包含与SEQ ID NO:5所示核苷酸序列差异不超过10、9、8、7、6、5、4、3、2或不超过1个核苷酸的核苷酸序列;
(D)具有SEQ ID NO:5所示核苷酸序列的,包括取代、缺失和/或插入一个或多个核苷酸的核苷酸序列。
在本发明的一个具体实施方式中,所述的人源化IL21R基因的核苷酸序列转录的mRNA选自下列组中的一种:
(a)包含SEQ ID NO:8所示核苷酸序列的全部或部分;
(b)包含与SEQ ID NO:8所示核苷酸序列的同一性至少为75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%或至少99%的核苷酸序列;
(c)包含与SEQ ID NO:8所示的核苷酸序列差异不超过10、9、8、7、6、5、4、3、2或不超过1个核苷酸的核苷酸序列;或
(d)包含SEQ ID NO:8所示的核苷酸序列所示的,包括取代、缺失和/或插入一个或多个核苷酸的核苷酸序列。
本发明的第十四方面,涉及一种表达上述的人源化IL21R蛋白的构建体。
本发明的第十五方面,涉及一种包含上述构建体的细胞。
本发明的第十六方面,涉及一种包含上述细胞的组织。
本发明的第十七方面,涉及了一种上述的非人动物、上述的荷瘤动物模型、上述的细胞或细胞系或原代细胞培养物、上述的组织或器官或其培养物、上述的人源化IL21R蛋白或上述的人源化IL21R基因在制备治疗或预防肿瘤的药物中的应用。
本发明的第十八方面,涉及一种上述的非人动物、上述的荷瘤动物模型、上述的细胞或细胞系或原代细胞培养物、上述的组织或器官或其培养物、上述的人源化IL21R蛋白或上述的人源化IL21R基因在IL21R基因或蛋白中相关研究中的应用,所述的应用包括:
A)涉及人类细胞的免疫过程的产品开发,制造或筛选人类抗体中的应用;
B)作为药理学、免疫学、微生物学和医学研究的模型系统中的应用;
C)涉及人类细胞的免疫过程的生产和利用动物实验疾病模型,用于病原学研究、用于开发诊断策略或用于开发治疗策略中的应用;
D)在体内研究人IL21R信号通路调节剂的筛选、药效检测、评估疗效、验证或评价;或者,
E)研究IL21R基因功能,研究人IL21R抗体,研究针对人IL21R靶位点的药物、药效,研究免疫相关疾病药物以及抗肿瘤或炎症药物方面的用途。
优选的,所述应用包括在制备药物组合物或者检测试剂盒中的用途。
优选的,所述应用不是疾病的诊断和治疗方法。
本发明所述的“肿瘤”包括但不限于淋巴瘤、B细胞肿瘤、T细胞肿瘤、骨髓/单核细胞肿瘤、非小细胞肺癌、白血病、卵巢癌、鼻咽癌、乳腺癌、子宫内膜癌、结肠癌、直肠癌、胃癌、膀胱癌、肺癌、支气管癌、骨癌、前列腺癌、胰腺癌、肝和胆管癌、食管癌、肾癌、甲状腺癌、头颈部癌、睾丸癌、胶质母细胞瘤、星形细胞瘤、黑色素瘤、骨髓增生异常综合征、以及肉瘤。其中,所述的白血病选自急性淋巴细胞性(成淋巴细胞性)白血病、急性骨髓性白血病、髓性白血病、慢性淋巴细胞性白血病、多发性骨髓瘤、浆细胞白血病、以及慢性骨髓性白血病;所述淋巴瘤选自霍奇金淋巴瘤和非霍奇金淋巴瘤,包括B细胞淋巴瘤、弥漫性大B细胞淋巴瘤、滤泡性淋巴瘤、套细胞淋巴瘤、边缘区B细胞淋巴瘤、T细胞淋巴瘤、和瓦尔登斯特伦巨球蛋白血症;所述肉瘤选自骨肉瘤、尤文肉瘤、平滑肌肉瘤、滑膜肉瘤、软组织肉瘤、血管肉瘤、脂肪肉瘤、纤维肉瘤、横纹肌肉瘤、及软骨肉瘤。在本发明的一个具体实施方式中,所述的肿瘤选自B细胞肿瘤、T细胞肿瘤、骨髓/单核细胞肿瘤。优选包括B或T细胞急性淋巴细胞白血病(ALL)、急性髓细胞白血病(AML)、非霍奇金淋巴瘤(NHL)和多发性骨髓瘤(MM)、鼻咽癌、肺癌。
本发明所述的“免疫相关疾病”包括但不限于过敏、哮喘、心肌炎、肾炎、肝炎、系统性红斑狼疮、类风湿性关节炎、硬皮病、甲状腺功能亢进、原发性血小板减少性紫癜、自身免疫性溶血性贫血、溃疡性结肠炎、自身免疫性肝病、糖尿病、疼痛或神经障碍等。在本发明的一个具体实施方式中。所述的免疫相关疾病为类风湿性关节炎。
本发明所述的“炎症”包括急性炎症,也包括慢性炎症。具体的,包括但不限于变质性炎症、渗出性炎症(浆液性炎、纤维素性炎、化脓性炎、出血性炎、坏死性炎、卡他性炎)、增生性炎症、特异性炎症(结核、梅毒、麻疯、淋巴肉芽肿等)。
本发明所述的IL21R基因人源化的非人动物体内可以正常表达人或人源化IL21R蛋白。可用于针对人IL21R靶位点的药物筛选、药效评估、免疫相关疾病和肿瘤治疗,可以加快新药研发过程、节约时间和成本。对于研究IL21R蛋白功能及相关疾病药物筛选提供了有效的保障。
本发明所述的“全部或部分”,“全部”为整体,“部分”为整体中的局部,或者组成整体的个体。
本发明所述的“人源化IL21R蛋白”,包含来源于人IL21R蛋白的部分和非人IL21R蛋白的部分。其中,所述的“人IL21R蛋白”同“人IL21R蛋白的全部”,即其氨基酸序列与人IL21R蛋白的全长氨基酸序列一致。所述的“人IL21R蛋白的部分”,为连续或间隔的5-538个(优选为10-188个)氨基酸序列与人IL21R蛋白的氨基酸序列一致或与人IL21R蛋白的氨基酸序列具有70%以上同源性。
本发明所述的“人IL21R蛋白的胞外区的全部”,代表其氨基酸序列分别与人IL21R蛋白的胞外区的全长氨基酸序列一致。
本发明所述的“人IL21R蛋白的胞外区的部分”,为连续或间隔5-213个(优选为5-188个)氨基酸序列与人IL21R蛋白的胞外区氨基酸序列一致,或与人IL21R蛋白的胞外区氨基酸序列具有70%以上同源性。
本发明所述的“人源化IL21R基因”,包含来源于人IL21R核苷酸序列的部分和非人IL21R基因的部分。其中,所述的“人IL21R核苷酸序列”同“人IL21R核苷酸序列的全部”,即其核苷酸序列与人IL21R核苷酸序列的全长核苷酸序列一致。所述的“人IL21R核苷酸序列的部分”为连续或间隔的20-49882bp(优选为20-10301bp或20-564bp)个核苷酸序列与人IL21R核苷酸序列一致或与人IL21R核苷酸序列具有70%以上同源性。
本发明所述的“xx号至xxx号外显子”或“xx号至xxx号外显子的全部”包含外显子及其期间的内含子的核苷酸序列,例如所述的“3号至6号外显子”包含3号外显子、3-4号内含子、4号外显子、4-5号内含子、5号外显子、5-6号内含子、6号外显子的全部核苷酸序列。
本发明所述的“x-xx号内含子”表示x号外显子与xx号外显子之间的内含子。例如“3-4号内含子”表示3号外显子与4号外显子之间的内含子。
本发明所述的“外显子的部分”表示连续或间隔几个、几十个或几百个核苷酸序列与全部的外显子核苷酸序列一致。例如人IL21R核苷酸序列的3号外显子的部分,包含连续或间隔的5-103bp个,优选10-83bp个核苷酸序列与人IL21R核苷酸序列的3号外显子核苷酸序列一致。在本发明的一个具体实施方式中,所述的“人源化IL21R基因”中包含的“3号外显子的部分”至少包括从3号外显子编码胞外区N端1-5(例如1、2、3、4、5)个氨基酸的核苷酸序列开始至3号外显子的最后一个核苷酸序列为止。
本发明所述的“基因座”广义上讲代表基因在染色体上所占的位置,狭义上讲代表某一基因上的一段DNA片段,即可以是一个基因也可以是一个基因的一部分。例如所述的“IL21R基因座”表示IL21R基因1号至9号外显子上的任选一段的DNA片段。优选为1号外显子、2号外显子、3号外显子、4号外显子、5号外显子、6号外显子、7号外显子、8号外显子、9号外显子或其期间的内含子中的任一个或两个或多个的组合,或一个或两个或多个的全部或部分,更优选为IL21R基因的3号至6号外显子上。
本发明所述的“核苷酸序列”包含天然的或经过修饰的核糖核苷酸序列、脱氧核糖核苷酸序列。优选为DNA、cDNA、pre-mRNA、mRNA、rRNA、hnRNA、miRNAs、scRNA、snRNA、siRNA、sgRNA、tRNA。
本发明所述“治疗(treating)”(或“治疗(treat)”或“治疗(treatment)”)表示减缓、中断、阻止、控制、停止、减轻、或逆转一种体征、症状、失调、病症、或疾病的进展或严重性,但不一定涉及所有疾病相关体征、症状、病症、或失调的完全消除。术语“治疗(treating)”等是指在疾病已开始发展后改善疾病或病理状态的体征、症状等等的治疗干预。
本发明所述“同源性”,是指在使用蛋白序列或核苷酸序列的方面,本领域技术人员可以根据实际工作需要对序列进行调整,使使用序列与现有技术获得的序列相比,具有(包括但不限于)1%,2%,3%,4%,5%,6%,7%,8%,9%,10%,11%,12%,13%,14%,15%,16%,17%,18%,19%,20%,21%,22%,23%,24%,25%,26%,27%,28%,29%,30%,31%,32%,33%,34%,35%,36%,37%,38%,39%,40%,41%,42%,43%,44%,45%,46%,47%,48%,49%,50%,51%,52%,53%,54%,55%,56%,57%,58%,59%,60%,70%,80%,81%,82%,83%,84%,85%,86%,87%,88%,89%,90%,91%,92%,93%,94%,95%,96%,97%,98%,99%,99.1%,99.2%,99.3%,99.4%,99.5%,99.6%,99.7%,99.8%,99.9%的同一性。
本领域的技术人员能够确定并比较序列元件或同一性程度,以区分另外的小鼠和人序列。
在一个方面,所述非人动物是哺乳动物。在一个方面,所述非人动物是小型哺乳动物,例如跳鼠科或鼠总科超家族。在一个实施方式中,所述基因修饰的动物是啮齿动物。在一个实施方式中,所述啮齿动物选自小鼠、大鼠和仓鼠。在一个实施方式中,所述啮齿动物选自鼠家族。在一个实施方式中,所述基因修饰的动物来自选自丽仓鼠科(例如小鼠样仓鼠)、仓鼠科(例如仓鼠、新世界大鼠和小鼠、田鼠)、鼠总科(真小鼠和大鼠、沙鼠、刺毛鼠、冠毛大鼠)、马岛鼠科(登山小鼠、岩小鼠、有尾大鼠、马达加斯加大鼠和小鼠)、刺睡鼠科(例如多刺睡鼠)和鼹形鼠科(例如摩尔大鼠、竹大鼠和鼢鼠)家族。在一个特定实施方式中,所述基因修饰的啮齿动物选自真小鼠或大鼠(鼠总科)、沙鼠、刺毛鼠和冠毛大鼠。在一个实施方式中,所述基因修饰的小鼠来自鼠科家族成员。在一个实施方式中,所述动物是啮齿动物。在一个特定实施方式中,所述啮齿动物选自小鼠和大鼠。在一个实施方式中,所述非人动物是小鼠。
在一个特定实施方式中,所述非人动物是啮齿动物,其为选自BALB/c、A、A/He、A/J、A/WySN、AKR、AKR/A、AKR/J、AKR/N、TA1、TA2、RF、SWR、C3H、C57BR、SJL、C57L、DBA/2、KM、NIH、ICR、CFW、FACA、C57BL/A、C57BL/An、C57BL/GrFa、C57BL/KaLwN、C57BL/6、C57BL/6J、C57BL/6ByJ、C57BL/6NJ、C57BL/10、 C57BL/10ScSn、C57BL/10Cr和C57BL/Ola的C57BL、C58、CBA/Br、CBA/Ca、CBA/J、CBA/st、CBA/H品系的小鼠。
除非特别说明,本发明的实践将采取细胞生物学、细胞培养、分子生物学、转基因生物学、微生物学、重组DNA和免疫学的传统技术。这些技术在以下文献中进行了详细的解释。例如:Molecular Cloning A Laboratory Manual,2ndEd.,ed. By Sambrook,FritschandManiatis (Cold Spring Harbor Laboratory Press:1989);DNA Cloning,Volumes I and II (D.N.Glovered.,1985);Oligonucleotide Synthesis (M.J.Gaited.,1984);Mullisetal. U.S. Pat.No.4,683,195;Nucleic Acid Hybridization(B.D.Hames& S.J.Higginseds.1984);Transcription And Translation (B.D.Hames&S.J.Higginseds.1984);Culture Of Animal Cells (R.I.Freshney,AlanR.Liss,Inc.,1987);Immobilized Cells And Enzymes (IRL Press,1986);B.Perbal,A PracticalGuide To Molecular Cloning(1984);the series,Methods In ENZYMOLOGY (J.Abelsonand M.Simon,eds.inchief,Academic Press,Inc.,New York),specifically,Vols. 154and 155 (Wuetal.eds.) and Vol.185,″Gene Expression Technology″ (D.Goeddel,ed.);Gene Transfer Vectors For Mammalian Cells (J.H.Miller and M.P.Caloseds.,1987,Cold Spring Harbor Laboratory);Immunochemical Methods In Cell AndMolecular Biology (Mayer and Walker,eds.,Academic Press,London,1987);HandbookOf Experimental Immunology,Volumes V (D.M.Weir and C.C.Blackwell,eds.,1986);and Manipulating the Mouse Embryo,(Cold Spring Harbor Laboratory Press,ColdSpring Harbor,N.Y.,1986)。
以上只是概括了本发明的一些方面,不是也不应该认为是在任何方面限制本发明。
本说明书提到的所有专利和出版物都是通过参考文献作为整体而引入本发明的。本领域的技术人员应认识到,对本发明可作某些改变并不偏离本发明的构思或范围。
下面的实施例进一步详细说明本发明,不能认为是限制本发明或本发明所说明的具体方法的范围。
附图说明
以下,结合附图来详细说明本发明的实施例,其中:
图1:人和小鼠IL21R基因结构对比示意图(非按比例);
图2:人源化IL21R基因座示意图(非按比例);
图3:IL21R基因打靶策略示意图(非按比例)
图4:重组后ES细胞Southern blot结果,其中WT为野生型对照;
图5:人源化IL21R小鼠FRT重组过程示意图(非按比例)
图6:F1代小鼠PCR结果,其中WT为野生型对照,H2O为水对照,PC为阳性对照;
图7:人IL21R和鼠IL21R的流式检测结果,其中WT为野生型C57BL/6小鼠,H/H为IL21R人源化纯合子小鼠。
具体实施方式
下面结合具体实施例来进一步描述本发明,本发明的优点和特点将会随着描述而更为清楚。但这些实施例仅是范例性的,并不对本发明的范围构成任何限制。本领域技术人员应该理解的是,在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换,但这些修改和替换均落入本发明的保护范围内。
在下述每一实施例中,设备和材料是从以下所指出的几家公司获得:
PerCP/Cy5.5 anti-mouse TCR β chainAntibody购自Biolegend,货号:553174;
Brilliant Violet 510™ anti-mouse CD45Antibody购自Biolegend,货号:103138;
PE anti-mouse IL-21R Antibody购自Biolegend,货号:131905;
APC anti-human CD360 (IL-21R) Antibody购自Biolegend,货号:347807;
FITC anti-Mouse CD19Antibody购自Biolegend,货号:115506;
Zombie NIR™ Fixable Viability Kit购自Biolegend,货号:423106;
PE Rat IgG2b, k isotype CtrlAntibody购自Biolegend,货号:101302。
实施例1 IL21R基因人源化小鼠的制备
本实施例对非人动物(如小鼠)进行改造,使该非人动物体内包含编码人源化IL21R蛋白的核苷酸序列,得到经遗传修饰的非人动物体内可表达人源化IL21R蛋白。小鼠IL21R基因(NCBI Gene ID:60504,Primary source:MGI:1890475,UniProt ID:Q9JHX3,位于7号染色体NC_000073.7的第125202424位至第125232742位,基于转录本NM_021887.2及其编码蛋白NP_ 068687.1(SEQ ID NO:1))和人IL21R基因(NCBI Gene ID:50615,Primarysource:HGNC:6006,UniProt ID:Q9HBE5,位于16号染色体NC_000073.7的第27402162-27452043位,基于转录本NM_181078.3及其编码蛋白NP_851564.1(SEQ ID NO:2))。对比示意图如图1所示。
为了达到本发明的目的,可在小鼠内源IL21R基因座引入编码人IL21R蛋白的基因序列,使得该小鼠表达人或人源化IL21R蛋白。具体来说,可以通过基因编辑技术在小鼠内源IL21R基因座上用人IL21R基因的核苷酸序列(例如DNA序列、cDNA序列等)替换小鼠相应序列,如将至少包含小鼠IL21R基因的3号外显子的部分序列至6号外显子的部分序列用对应的人DNA序列替换,得到人源化IL21R基因座(示意图如图2所示),实现对小鼠IL21R基因的人源化改造。
进一步设计如图3所示的打靶策略示意图,图中显示了靶向载体上含有小鼠IL21R基因上游和下游的同源臂序列,以及包含人IL21RDNA序列的A片段。其中,上游同源臂序列(5’同源臂,SEQ ID NO:3)与NCBI登录号为NC_000073.7第125219443至125224456位核苷酸序列相同,下游同源臂序列(3’同源臂,SEQ ID NO:4)与NCBI登录号为NC_000073.7第125230780至125235238位核苷酸序列相同;人IL21R DNA序列(SEQ ID NO:5)与NCBI登录号为NC_000016.10的第27434367至27444667位核苷酸序列相同。
靶向载体上还包括用于阳性克隆筛选的抗性基因,即新霉素磷酸转移酶编码序列Neo,并在抗性基因的两侧装上两个同向排列的位点特异性重组系统Frt重组位点,组成Neo盒(Neo cassette)。其中,Neo盒5’端与鼠的连接设计为5’-tctagctcaagaccacacagctaaggagtgactccaggtcaGATATCGAATTCCGAAGTTCCTATTCTCTAGAAAGTATAGGAACTTCAGGTCT-3’(SEQID NO:6),其中,序列“caggtca”的最后一个“a”是鼠的最后一个核苷酸,序列“GATATC”的第一个“G”是Neo盒的第一个核苷酸;Neo盒3’端与鼠的连接设计为5’-CTCTAGAAAGTATAGGAACTTCATCAGTCAGGTACATAATGGTGGATCCGATATCggggtagagagatgtagtggttctgagatgttctacggggtgatctctgccaat-3’(SEQ ID NO:7)内,其中序列“GATATC”的“C”是Neo盒的最后一个核苷酸,序列“ggggta”的第一个“g”是鼠的第一个核苷酸。此外,还在靶向载体3’同源臂下游构建了具有负筛选标记的编码基因(白喉毒素A亚基的编码基因(DTA))。改造后的人源化小鼠IL21R的mRNA序列如SEQ ID NO:8所示,表达的蛋白序列如SEQ ID NO:23所示。
靶向载体构建可采用常规方法进行,如酶切连接等。构建好的靶向载体通过酶切进行初步验证后,再送测序公司进行测序验证。将测序验证正确的靶向载体电穿孔转染入C57BL/6小鼠的胚胎干细胞中,利用阳性克隆筛选标记基因对得到的细胞进行筛选,并利用PCR(PCR引物详见表1)和Southern Blot技术进行检测确认外源基因的整合情况,筛选出正确的阳性克隆细胞,经PCR鉴定为阳性的克隆再进行Southern Blot(分别用BglII或SacI或AseI消化细胞DNA并使用3个探针进行杂交,酶、探针及目的片段长度如表2所示)检测,Southern Blot检测结果如图4所示,表明6个经PCR验证为阳性的胚胎干细胞(ES-01、ES-02、ES-03、ES-04、ES-05和ES-06)均为阳性的克隆,且无随机插入。
表1 PCR检测引物序列及目的片段长度
表2 Southern Blot酶和探针表
Southern Blot检测包括如下探针引物:
5’探针(5’Probe):
5’Probe-F:5’-TGTGCCTATGACTGTTGCAGGTGTT -3’(SEQ ID NO:13),
5’Probe-R:5’- CCTCGCTATCCTAACATGCCAGCTC-3’(SEQ ID NO:14);
3’探针(3’Probe):
3’Probe-F:5’- AATGAATAGGGTGTGCAGGGTGCAT-3’(SEQ ID NO:15),
3’Probe-R:5’-GCAGCATCCCCACAGAGAGAAACTA-3’(SEQ ID NO:16);
Neo探针(Neo Probe):
Neo Probe-F:5’-GGATCGGCCATTGAACAAGAT -3’(SEQ ID NO:17),
Neo Probe-R:5’-CAGAAGAACTCGTCAAGAAGGC -3’(SEQ ID NO:18)。
将筛选出的正确阳性克隆细胞(黑色鼠)按照本领域已知的技术导入已分离好的囊胚中(白色鼠),得到的嵌合囊胚转移至培养液中短暂培养后移植至受体母鼠(白色鼠)的输卵管,可生产F0代嵌合体鼠(黑白相间)。将F0代嵌合鼠与野生型鼠回交获得F1代鼠,再将F1代杂合小鼠互相交配即可获得F2代纯合子鼠。还可将阳性鼠与Flp工具鼠交配去除阳性克隆筛选标记基因(该过程示意图见图5)后,再通过互相交配即可得到人源化IL21R基因纯合子小鼠。示例性的F1代小鼠鉴定结果见图6,其中,编号为F1-01至F1-06的小鼠均为阳性杂合小鼠,PCR测定引物如表3所示。
表3 PCR检测引物序列及目的片段长度
进一步采用流式细胞术检测IL21R基因人源化小鼠体内IL21R蛋白的表达情况。选取9周龄野生型C57BL/6小鼠和IL21R基因人源化纯合子小鼠各1只,取脾脏细胞,用抗鼠CD45抗体Brilliant Violet 510™ anti-mouse CD45Antibody、鼠源T细胞表面抗体PerCP/Cy5.5 anti-mouse TCR β chain、抗鼠IL21R抗体anti-mouse IL-21R Antibody,PE, Biolegend™(mIL21R-PE)、( 以PE大鼠抗体IgG2a同型对照抗体Rat IgG2a, κIsotype Ctrl Antibody, PE, Biolegend™(IgG2a)抗体作参照),抗鼠CD19抗体FITCanti-Mouse CD19和抗人CD360抗体anti-human CD360 (IL-21R) Antibody ,APC,Biolegend™(hIL21R-APC)(以鼠抗体Mouse IgG1, κ Isotype Ctrl (FC) Antibody,APC,Biolegend™为参照)进行染色,将染色后的细胞进行流式检测,hIL21R在B细胞中的检测结果如图7所示,在生理状态下的野生C57BL/6小鼠(WT)B细胞中只检测到鼠IL21R蛋白,纯合鼠(H/H,Homozygote)B细胞中只检测到人源化IL21R蛋白。以上表明本实施例成功构建出可表达人源化IL21R蛋白的IL21R基因人源化小鼠。
实施例2 体内药效验证
利用本方法制得的IL21R人源化小鼠可以用于评估靶向人IL21R的调节剂的药效。例如,取IL21R人源化小鼠纯合子皮下接种小鼠结肠癌细胞MC38,待肿瘤体积生长到约100mm3后根据肿瘤体积分为对照组或治疗组,治疗组随机选择靶向人IL21R的药物X、Y、Z等,对照组注射等体积的生理盐水。定期测量肿瘤体积并称量小鼠的体重,可通过比较小鼠体重变化和肿瘤大小即可有效评估化合物的体内安全性和体内药效。
实施例3双基因或多基因人源化小鼠
利用本方法或制得的IL21R基因人源化小鼠还可以制备双基因修饰或多基因修饰的小鼠模型。如前述实施例1中,囊胚显微注射使用的胚胎干细胞可选择来源于含有PD-1、PD-L1、CTLA4、OX40、LAG3、TIM3、CD73等其它基因修饰的小鼠,或者,也可在人源化IL21R小鼠的基础上,利用分离小鼠ES胚胎干细胞和基因重组打靶技术,获得IL21R与其它基因修饰的双基因或多基因修饰的小鼠模型。也可将本方法得到的IL21R小鼠纯合子或杂合子与其它基因修饰的纯合或杂合小鼠交配,对其后代进行筛选,根据孟德尔遗传规律,可有一定机率得到IL21R基因与其它基因修饰的双基因或多基因修饰的杂合小鼠,再将杂合子相互交配可以得到双基因或多基因修饰的纯合子,利用这些双基因或多基因修饰的小鼠可以进行靶向人IL21R和其它基因调节剂的体内药效验证等。
以上详细描述了本发明的优选实施方式,但是,本发明并不限于上述实施方式中的具体细节,在本发明的技术构思范围内,可以对本发明的技术方案进行多种简单变型,这些简单变型均属于本发明的保护范围。
另外需要说明的是,在上述具体实施方式中所描述的各个具体技术特征,在不矛盾的情况下,可以通过任何合适的方式进行组合,为了避免不必要的重复,本发明对各种可能的组合方式不再另行说明。
此外,本发明的各种不同的实施方式之间也可以进行任意组合,只要其不违背本发明的思想,其同样应当视为本发明所公开的内容。
序列表
<110> 百奥赛图(北京)医药科技股份有限公司
<120> IL21R基因人源化非人动物及其构建方法和应用
<130> 1
<160> 24
<170> SIPOSequenceListing 1.0
<210> 1
<211> 529
<212> PRT
<213> 小鼠(Mouse)
<400> 1
Met Pro Arg Gly Pro Val Ala Ala Leu Leu Leu Leu Ile Leu His Gly
1 5 10 15
Ala Trp Ser Cys Leu Asp Leu Thr Cys Tyr Thr Asp Tyr Leu Trp Thr
20 25 30
Ile Thr Cys Val Leu Glu Thr Arg Ser Pro Asn Pro Ser Ile Leu Ser
35 40 45
Leu Thr Trp Gln Asp Glu Tyr Glu Glu Leu Gln Asp Gln Glu Thr Phe
50 55 60
Cys Ser Leu His Arg Ser Gly His Asn Thr Thr His Ile Trp Tyr Thr
65 70 75 80
Cys His Met Arg Leu Ser Gln Phe Leu Ser Asp Glu Val Phe Ile Val
85 90 95
Asn Val Thr Asp Gln Ser Gly Asn Asn Ser Gln Glu Cys Gly Ser Phe
100 105 110
Val Leu Ala Glu Ser Ile Lys Pro Ala Pro Pro Leu Asn Val Thr Val
115 120 125
Ala Phe Ser Gly Arg Tyr Asp Ile Ser Trp Asp Ser Ala Tyr Asp Glu
130 135 140
Pro Ser Asn Tyr Val Leu Arg Gly Lys Leu Gln Tyr Glu Leu Gln Tyr
145 150 155 160
Arg Asn Leu Arg Asp Pro Tyr Ala Val Arg Pro Val Thr Lys Leu Ile
165 170 175
Ser Val Asp Ser Arg Asn Val Ser Leu Leu Pro Glu Glu Phe His Lys
180 185 190
Asp Ser Ser Tyr Gln Leu Gln Val Arg Ala Ala Pro Gln Pro Gly Thr
195 200 205
Ser Phe Arg Gly Thr Trp Ser Glu Trp Ser Asp Pro Val Ile Phe Gln
210 215 220
Thr Gln Ala Gly Glu Pro Glu Ala Gly Trp Asp Pro His Met Leu Leu
225 230 235 240
Leu Leu Ala Val Leu Ile Ile Val Leu Val Phe Met Gly Leu Lys Ile
245 250 255
His Leu Pro Trp Arg Leu Trp Lys Lys Ile Trp Ala Pro Val Pro Thr
260 265 270
Pro Glu Ser Phe Phe Gln Pro Leu Tyr Arg Glu His Ser Gly Asn Phe
275 280 285
Lys Lys Trp Val Asn Thr Pro Phe Thr Ala Ser Ser Ile Glu Leu Val
290 295 300
Pro Gln Ser Ser Thr Thr Thr Ser Ala Leu His Leu Ser Leu Tyr Pro
305 310 315 320
Ala Lys Glu Lys Lys Phe Pro Gly Leu Pro Gly Leu Glu Glu Gln Leu
325 330 335
Glu Cys Asp Gly Met Ser Glu Pro Gly His Trp Cys Ile Ile Pro Leu
340 345 350
Ala Ala Gly Gln Ala Val Ser Ala Tyr Ser Glu Glu Arg Asp Arg Pro
355 360 365
Tyr Gly Leu Val Ser Ile Asp Thr Val Thr Val Gly Asp Ala Glu Gly
370 375 380
Leu Cys Val Trp Pro Cys Ser Cys Glu Asp Asp Gly Tyr Pro Ala Met
385 390 395 400
Asn Leu Asp Ala Gly Arg Glu Ser Gly Pro Asn Ser Glu Asp Leu Leu
405 410 415
Leu Val Thr Asp Pro Ala Phe Leu Ser Cys Gly Cys Val Ser Gly Ser
420 425 430
Gly Leu Arg Leu Gly Gly Ser Pro Gly Ser Leu Leu Asp Arg Leu Arg
435 440 445
Leu Ser Phe Ala Lys Glu Gly Asp Trp Thr Ala Asp Pro Thr Trp Arg
450 455 460
Thr Gly Ser Pro Gly Gly Gly Ser Glu Ser Glu Ala Gly Ser Pro Pro
465 470 475 480
Gly Leu Asp Met Asp Thr Phe Asp Ser Gly Phe Ala Gly Ser Asp Cys
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Gly Ser Pro Val Glu Thr Asp Glu Gly Pro Pro Arg Ser Tyr Leu Arg
500 505 510
Gln Trp Val Val Arg Thr Pro Pro Pro Val Asp Ser Gly Ala Gln Ser
515 520 525
Ser
<210> 2
<211> 538
<212> PRT
<213> 人(human)
<400> 2
Met Pro Arg Gly Trp Ala Ala Pro Leu Leu Leu Leu Leu Leu Gln Gly
1 5 10 15
Gly Trp Gly Cys Pro Asp Leu Val Cys Tyr Thr Asp Tyr Leu Gln Thr
20 25 30
Val Ile Cys Ile Leu Glu Met Trp Asn Leu His Pro Ser Thr Leu Thr
35 40 45
Leu Thr Trp Gln Asp Gln Tyr Glu Glu Leu Lys Asp Glu Ala Thr Ser
50 55 60
Cys Ser Leu His Arg Ser Ala His Asn Ala Thr His Ala Thr Tyr Thr
65 70 75 80
Cys His Met Asp Val Phe His Phe Met Ala Asp Asp Ile Phe Ser Val
85 90 95
Asn Ile Thr Asp Gln Ser Gly Asn Tyr Ser Gln Glu Cys Gly Ser Phe
100 105 110
Leu Leu Ala Glu Ser Ile Lys Pro Ala Pro Pro Phe Asn Val Thr Val
115 120 125
Thr Phe Ser Gly Gln Tyr Asn Ile Ser Trp Arg Ser Asp Tyr Glu Asp
130 135 140
Pro Ala Phe Tyr Met Leu Lys Gly Lys Leu Gln Tyr Glu Leu Gln Tyr
145 150 155 160
Arg Asn Arg Gly Asp Pro Trp Ala Val Ser Pro Arg Arg Lys Leu Ile
165 170 175
Ser Val Asp Ser Arg Ser Val Ser Leu Leu Pro Leu Glu Phe Arg Lys
180 185 190
Asp Ser Ser Tyr Glu Leu Gln Val Arg Ala Gly Pro Met Pro Gly Ser
195 200 205
Ser Tyr Gln Gly Thr Trp Ser Glu Trp Ser Asp Pro Val Ile Phe Gln
210 215 220
Thr Gln Ser Glu Glu Leu Lys Glu Gly Trp Asn Pro His Leu Leu Leu
225 230 235 240
Leu Leu Leu Leu Val Ile Val Phe Ile Pro Ala Phe Trp Ser Leu Lys
245 250 255
Thr His Pro Leu Trp Arg Leu Trp Lys Lys Ile Trp Ala Val Pro Ser
260 265 270
Pro Glu Arg Phe Phe Met Pro Leu Tyr Lys Gly Cys Ser Gly Asp Phe
275 280 285
Lys Lys Trp Val Gly Ala Pro Phe Thr Gly Ser Ser Leu Glu Leu Gly
290 295 300
Pro Trp Ser Pro Glu Val Pro Ser Thr Leu Glu Val Tyr Ser Cys His
305 310 315 320
Pro Pro Arg Ser Pro Ala Lys Arg Leu Gln Leu Thr Glu Leu Gln Glu
325 330 335
Pro Ala Glu Leu Val Glu Ser Asp Gly Val Pro Lys Pro Ser Phe Trp
340 345 350
Pro Thr Ala Gln Asn Ser Gly Gly Ser Ala Tyr Ser Glu Glu Arg Asp
355 360 365
Arg Pro Tyr Gly Leu Val Ser Ile Asp Thr Val Thr Val Leu Asp Ala
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Glu Gly Pro Cys Thr Trp Pro Cys Ser Cys Glu Asp Asp Gly Tyr Pro
385 390 395 400
Ala Leu Asp Leu Asp Ala Gly Leu Glu Pro Ser Pro Gly Leu Glu Asp
405 410 415
Pro Leu Leu Asp Ala Gly Thr Thr Val Leu Ser Cys Gly Cys Val Ser
420 425 430
Ala Gly Ser Pro Gly Leu Gly Gly Pro Leu Gly Ser Leu Leu Asp Arg
435 440 445
Leu Lys Pro Pro Leu Ala Asp Gly Glu Asp Trp Ala Gly Gly Leu Pro
450 455 460
Trp Gly Gly Arg Ser Pro Gly Gly Val Ser Glu Ser Glu Ala Gly Ser
465 470 475 480
Pro Leu Ala Gly Leu Asp Met Asp Thr Phe Asp Ser Gly Phe Val Gly
485 490 495
Ser Asp Cys Ser Ser Pro Val Glu Cys Asp Phe Thr Ser Pro Gly Asp
500 505 510
Glu Gly Pro Pro Arg Ser Tyr Leu Arg Gln Trp Val Val Ile Pro Pro
515 520 525
Pro Leu Ser Ser Pro Gly Pro Gln Ala Ser
530 535
<210> 3
<211> 5014
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
ggagaacagc aggagtgggt gtggcttaca ccggataccc gtcctgccac tggctgctct 60
cagctctgag gttcaggggt agaatgtatt catgtcaaag atggcaactg aggccctggt 120
catccacaaa aagatcagag gtcagctgaa tactgggtcc ctcctcctct ttgctatgcc 180
aggaaaaagc acacagaaat ataactaaca cagtataggg gggatagggg tctttgggct 240
catgtaaatg aaaggtttac atgtgggcat cagatataac ctaattcatg ttcttaatgt 300
agccctgctg ttgtggctcg aatctgaagt gtctcctata ggctcatgtg tttaaatgct 360
tgttcctacg attggggacg gagcctggat gaaggaagta ggtctctggg ggcgggcctt 420
tgagggtgat acccaccctg tttcctgcct tgctccctgc ttggtgggcc atcacgatgg 480
gaatggcctc tgccaagcac ctgtatcacc actctaccac accttcctgt tgggacaggc 540
tgaacccctg aaatcatgag ccaaggtaca actctaagag acaaccaaag gtgtctgcag 600
gtttctgata ggcgcccaga aaagcatcaa tatcaggaat ctacagatga ggccaggctg 660
tctgccactt gcttccctga gcttgtgtct tgctgggcct ggttctgtgc cttgtctgtg 720
gaccatgcag aggggctgca gctgagagag cccaccgtga ggagggctgc caattcaggc 780
agacagagca gctgatgtct gtctgtggct tgtgtagagc tgtggtgtgg agtggaagag 840
cggaggctcc tgacaaagga cggaagggta tccactttcc ctctggctga ggttcgacac 900
agagttctgc tctgctggga tgtctgtcgc aggtgtgaca ccttagagag gagcaggctt 960
ggagcaagcc tcctatggtc agagctcacc tcttcagagt gtcatatcag gggtggggga 1020
gggggctttg ctcagtctgg tgctgcacca aagcaccgga ccctggcaag tgagaaggat 1080
gtttattcag ctgagggtct agcatctggg gagccagggc catggccact atcacagcct 1140
ggtagggagc tgtacaagga ggcacagctg atgcgccagt ttggatctct cttcctttcc 1200
tcccatgtcc tccctgcttg cctttctttt ccaatgctgg gggcagaaac cagggactca 1260
tatatgttta ggcaaatact ggaataccga gttccattcc tgccatgatg tattcaaaac 1320
tcactaatgc catcacgggc tctgccttca tgacctcagc taatcccgat tccctccatt 1380
ctattaaagt atggtctggg tgttaggttt ctaacacaca aagttttttt tgggggggtg 1440
tgtgtgacac aaaccatacc acagggcttg tgaaggtggg ggccacccaa gatcagggtt 1500
actcaaagca ggttaactga cattttccat aattttttta ttctaattct gatgttatag 1560
taacgactat aattacaata gttataaggc tgaagcagga agttcagaag gtcatgatca 1620
gtcttggtta catagttaag actctgtctc aagaagacag gggtggaaag gaagaaaata 1680
cacccctgtg gaaaaagtgt acaccacgag tgtgaatttt tagaatgggg ttggaataaa 1740
attaagcatt tatagaaaaa cagtaaataa gtaattttga aagtgaagca ttttggaagc 1800
cccttgggca cagcgttcct ggttgttctg ttccagaact ttctggactc tgttctagct 1860
cagtcagccc cgcacatgct gtgtgtcaga gcctatgccc agctttctct acacaaagct 1920
ctagacaggc cttcattgct tcattttatg aggcccagag atagcaagag actggcctgg 1980
ggccacacag ctggccttct gaaccttggt catttacact gcttctttga tgatggtctg 2040
gtaggctact caatgctgct ggatttgggg gaagcccgga tgaaatgtag tcttatggtg 2100
aggtctggga tagtgtctga cctgacccat gcgtaggcta ccttccttca aggccctggt 2160
ccttgctcca cagtagaaga gctggactgc acgagactcc tgagcaggtg gtctccctga 2220
gctagcactt ctataggcaa agggcaggag gagcctggtg ttgtccacct atggttaccc 2280
ttctgcccac agcatcatta cccccttgat tctacagggg ccctgtctat ctcagccagt 2340
caggggctct cagcaacttt gtgtggtgcc ctggctttcc tagtgtgcca ctgtgggcta 2400
ccccgggctt cctgttctaa taagaatacc tcctaggtcc cccatgggct aacctcatct 2460
ttggtactca acaggggtct tctttatgag cttcggacca gctcttttga tgtggcaggg 2520
actgaccctg ggtggggaag ccactcagtg catgacccca gctggttcac cacatatacc 2580
acatactttt cttgcaggtc tgggacacag catgccccgg ggcccagtgg ctgccttact 2640
cctgctgatt ctccatggag gtaagcctca gtaccatgac tgcaggatgg atggaggcca 2700
tctcctccca caaagctcag ctgggcccac atgatgttgt aggctcctgg tcatgtcatg 2760
gctgctggcc aacaacccac aatccacagc ctctgtctat ttgaccctaa aactgaatgc 2820
ctcatttcca aagagctcca gtaaacagaa aagctctcca ccatcatggg aaggaagtca 2880
tacccttggt ctaacctcca ttcctctttg tgcaagactc ttactcaggt catgaaccct 2940
cagtaggctg gaggagaagg gaggtggaga tagagaagcc agaattctgt tggcagtccc 3000
tgaggaaagg cgtcagaact gcaggccaag agtcctgttt agggcactct ccccaaagcc 3060
aagagtccag aagtgctctt cctcaagagc taggagatga gaggctgttt agactgccca 3120
gggcgagggg aacttgactc cctgcaggat cagcaccatt tgctctgggt tttaccgaaa 3180
tatccagtac tttagttccc ttaggaaaca ggcgtgactc ttgatttccc tttctgtttg 3240
cgacaccctg cctgctctgg aaacccctgt cattcctgag tcagggagag gaggttgggg 3300
ggctgggtat caggcacttg tcacaaagac tagaatgtca tctcctgctc ttcctgtgga 3360
gcctggctgt gttgccatca gtggaaagac agaggttcag ggagaacaca gaccctgcgg 3420
ccaccctgtg ggtggctgcc tgactcttag gtctgccttt gtctcatggc actaagacaa 3480
aatataagga gcttggatag gatgatggct ttggtgatgt gcttgtcatt ccagcatcag 3540
gacccatgga agggacctag tgcagctgag catacttgca tggggtgtgg ggggggtggt 3600
ggaagcagat cccaggggca tgctggtcag gctgctcagc taaatcagtg agttgcaggc 3660
tgggcgagag agcctatctc aacaaacaag gttgagaaag actgagaaag acccctgtct 3720
gcaacttgca tgtgtgcatg catgtgcagg agcctacaca cagggtcaaa tgtctatctt 3780
tcgcagcctg ggtgctcaga cactctatgc ctggaaaagt ccagcatcta gttccggtgt 3840
ggtgctgacc ctcttgtttt cttgatggga gacaggatct gggcagacag aagtctggtc 3900
cctccagcca cttgtaacac cactaggccc ccttaggagc ttggtctggg gactctattg 3960
ctgtggaggc cccgcctcct cacatgctca gtggcctgag catttgatgg ggcacacaca 4020
ttctaatttt accagggccc caggtacctg ttgtgctgct attagttcat tcatttattc 4080
attcattttt gagctttgga gacaaggttt tggtctgtag ttgtaatcct cctgcctcag 4140
cttgctgcta tgtcaattct atgttatttt tatttcatag tcagatactt ttcctttccc 4200
tttaagacag gatctggctg tgtagcccag gctggtctta aattgggaat ccacctgcct 4260
ctgttcccca aatgcttggg aaggcagatg tgtcccatgc ctgctctact gtcatttgca 4320
aaggaaacaa atggctgggg tggtggggga gagggtggag gagaggttag gggagcaaga 4380
gggagagagg gtgggaaaga gggagggagg gggagaggga gggggagagg gggagggaga 4440
agatagagga gaaggtgggg gagagggaga gggtggggga gggggagggg gaaagggtgg 4500
gagagaggga gggggagagg tggctcttcc agcctagtgc ctgctgcagc attcaacctg 4560
gtagccatgc tgtcatcagc agcgacacag ttaacttttc tacaaactct gaagtcttct 4620
ccactctttc accggcttcc ctaacccctc ccctcctgga agaggctaca agctcatgtc 4680
cctgtgccac ttgccctgac ctgggggatt tatttgttct ctgcctatag tggattcaaa 4740
gtgttttacc ctatgaggga cagtggtctt agaaccaaat atcgagccca gtttcacatt 4800
aaaagagcct catcaccatg actactcttg aaatgttggg actcacacaa aaacttctag 4860
atgacaagag tccccgtgtc ccaggctttc atcccagcct gtgactctgc ccaagcttgt 4920
cctcagcctc acacaacctg ccaaggcctt tcccctgtgt ctcacccaca agggcctctc 4980
tgttatcccg ccagcttgga gctgcctgga cctc 5014
<210> 4
<211> 4459
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
ggggtagaga gatgtagtgg ttctgagatg ttctacgggg tgatctctgc caatgcagta 60
gtctctttta ttgtaagcaa tatcctgcag gaccactgtc tccagtccag catccccaaa 120
ggcctttcaa cacggctttt tagtaattca ttccatctat aaacatttat ggtacaccta 180
ctgtgtgcca ggtactgagg acacagttgt gatcagggct agtgtagaca cacaagcaaa 240
actagagaca tccggaagtg tcaggagacg gagtagaggc tgggccactt agacctcagg 300
ctctccctgc acacgtcctc aagaccttag gacttaggaa cctggtccca gcacccagct 360
gttccttggc tggggcactg gtaactagcg tggatatgag acagaggaca gtcagtcctt 420
actaaaggtg ggaacacggg ctctgagaac ggacagtatt gggaacccac tgggcagggg 480
gttcacagac agacatcatg gcgcgctctc tctctctctc tctctcctgt tttcttgttc 540
ttctgctttc cccgtctctg gcttgtccct gtactccccc ccccaccccc atctttggct 600
ctctctgttc acacccgacc ttgttgtccc cagctcatga ctgtgtgttt ctttctcata 660
gaaatgggtt aatacccctt tcacggcctc cagcatagag ttggtgccac agagttccac 720
aacaacatca gccttacatc tgtcattgta tccagccaag gagaagaagt tcccggggct 780
gccgggtctg gaagagcaac tggagtgtga tggaatgtct gagcctggtc actggtgcat 840
aatccccttg gcagctggcc aagcggtctc agcctacagt gaggagagag accggccata 900
tggtctggtg tccattgaca cagtgactgt gggagatgca gagggcctgt gtgtctggcc 960
ctgtagctgt gaggatgatg gctatccagc catgaacctg gatgctggcc gagagtctgg 1020
ccctaattca gaggatctgc tcttggtcac agaccctgct tttctgtctt gcggctgtgt 1080
ctcaggtagt ggtctcaggc ttggaggctc cccaggcagc ctactggaca ggttgaggct 1140
gtcatttgca aaggaagggg actggacagc agacccaacc tggagaactg ggtccccagg 1200
agggggctct gagagtgaag caggttcccc ccctggtctg gacatggaca catttgacag 1260
tggctttgca ggttcagact gtggcagccc cgtggagact gatgaaggac cccctcgaag 1320
ctatctccgc cagtgggtgg tcaggacccc tccacctgtg gacagtggag cccagagcag 1380
ctagcatata ataaccagct atagtgagaa gaggcctctg agcctggcat ttacagtgtg 1440
aacatgtagg ggtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtctt gggttgtgtg 1500
ttagcacatc catgttggga tttggtctgt tgctatgtat tgtaatgcta aattctctac 1560
ccaaagttct aggcctacga gtgaattctc atgtttacaa acttgctgtg taaaccttgt 1620
tccttaattt aataccattg gttaaataaa attggctgca accaattact ggagggatta 1680
gaggtagggg gcttttgagt tacctgtttg gagatggaga aggagagagg agagaccaag 1740
aggagaagga ggaaggagag gagaggagag gagaggagag gagaggagag gagaggagag 1800
gagaggagag gagaggctgc cgtgagggga gagggaccat gagcctgtgg ccaggagaaa 1860
cagcaagtat ctggggtaca ctggtgagga ggtggccagg ccagcagtta gaagagtaga 1920
ttaggggtga cctccagtat ttgtcaaagc caattaaaat aaccatagtg gagtcttatt 1980
tatttgtacg ctagtcaggg ataagtttaa aattggttgt actacacatc catttgtgag 2040
caatgcctct gcacgtccag tgttcactgt caatgcctgt acgtgtgtgc ctgggctcac 2100
gtgtgtatgc acgagtggag tgtaggtgtc tcttagatag gagcctctga ctcttcccct 2160
gcggacatgt ggggtggtca cactcagtgt caagtgattg gtccagattg ctgcccctgg 2220
ggcactatca ccaagctcga ccctaccagg ggccatctct aggacctatg aagtcctgac 2280
tatcttttat cccattagaa tgagaagatt tttggggaac ctagggatgt aagctgtaaa 2340
ccccagctct ggtggctcca ggctatgcag cctggagaag ccacttctga attctatgct 2400
gactttactt gcctggacat taggaatgac gtaggtaccc tggatcagct ctatacagtc 2460
tgcagcccca ttctgcttcc tgtcttttgt ggcttctagg atccaggtgc atactctggg 2520
gtgcatactc caggtgtgtg tgtgtgtgtg tgtgtataca tatatatata tatatatata 2580
tatatatata tatatatata tatacaccag cctacactac acccacacac ccacagcctg 2640
ggaaccagtg gggcatcctc ccaccaagag tactcagcag gtttccaggc tgtgacacca 2700
gaagcagccc caccaaccaa gttgggtttt cgtttgtttg tttgtttgtt ttctggcttc 2760
ttctgcccat taaaaagaaa atttctggga ggcggcgttg gcacatacct ctaatcctca 2820
tctctgactg caaggccaat ctggtataca gaatgagttc caggacaacc tgtgtcaaca 2880
aacaaacaaa caaacaaaca aacaaacata agataacaac aaaaaccaaa cccaaaccac 2940
caccaaaaaa caaaacaaaa caaaacaaaa caaaaacaaa ccaaccaaac aaaccaaaca 3000
aacaaacaac ctggagaagg aggaggagga ggaagaggag gaggaggagg aggaagagga 3060
ggaggaggaa gaggaggagg aggaggagga ggaggaggag gaggaagaag aggaggagga 3120
ggaagaggag gaggaggaag aggaggagga ggaagaggag gaggaagagg aagaggagga 3180
ggattctttt aagtatccag gaagtattta taatacaagg cacagggaga gggtacagag 3240
cacagtacct acagaacccc ctgtatgaca cccacagagg acaggggacc caaaggagcc 3300
atcaagaagc agctctggtt ctgtagctag tacctgtgtt ccccctaccc tcggtgccat 3360
gcttatccta ggttagggga ggttcccttg catccaatga ttcttttgag cacgatgttc 3420
gctgtgtgag ggatgctaag gtatgctggg aggtattgga gaactccttc ccagtcacca 3480
ggatacagta agagcgcgtg ggtgctggaa ttgcttagaa cttgctgttg ttttcttctc 3540
tccctccttc atttctgttt ccctcttccc ttcctttgtc gttctcttct cgaatggtct 3600
gttctccctg caaggatgca cctgagaggt ggggtggtat ttgcaaaacc aggcaggatg 3660
gaggcaacct ggtgggaagc ctccatagtg gctgggattc cggggtacag attctcccct 3720
caatggccca ggaaaaaagg ctgtctacct gtccagtctg cagagctgga ttggaaagaa 3780
gttagcagcc gcctgctcat ttgcaaggca gggtagctgg tggaatcacc aagggaatgc 3840
atgtatgtcc tccgtatctg actgagggct ggaaagttct ggaacactgg tagtttcgaa 3900
agtgagacag agcctaggac cagggtgcga gagagaagct cagagcagca aaacccccag 3960
ctctttcctg cacgtgcaca caaccccacc cggtgcaccc agacacttag accaaagtgt 4020
gctgtggttt tcacatactc tgggagccac agactgcggg tgctcttggc agctctccag 4080
ggaaccaact cagcccctca gcagggagcg ctcctgacag gggtgtgggc aggaagaaca 4140
ttcagggctg aggccctggc cctcaccagc tccatttgaa ggggcgcctc tcgggaacaa 4200
acgtggatgt ggaagttgaa ggcaagagct ggaccaccta gtagtggatg aaaggatggc 4260
ggaaggcttg agagaatggc ggctggatgc agagacggag atactgcaca agagcagccg 4320
accagatgca gccagtaaag ctagttagtt gctctgtctg ctctcagttt tacccccagc 4380
cctacacaca acgggtgctt aataaagtat tagcaaagcc cgatggtagc ctgtcttatt 4440
tccaggacct caggctcca 4459
<210> 5
<211> 10301
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
gtctgctaca ccgattacct ccagacggtc atctgcatcc tggaaatgtg gaacctccac 60
cccagcacgc tcacccttac ctggtaagta gccgggcctc accagtcccc ggggatgcaa 120
ttcagggtgc ctgctcagtg attcccccag gaggacactg tgcactgagg accactgtgt 180
ccgcctttca gacaaccact cagggctcac aggacttgaa aagtcaggag cccgcagggt 240
tggtctatcc accagcttca aaggccactc tctcacccac acaggtcggc catggttttg 300
agaccccggg atggctggtt tcccgcatga tttttcaagc acttaggtac caggccccca 360
actacacgct tcccatgaat ggtcaccccc aaccctcaca atgattactc ccattttaca 420
gatgaggaca ttgaggttca gagaggttaa gcaacttgcc taaggtcaca cagccagtga 480
atgctggagc tgggattcaa attcagaccg cccaactcca aagtccatgg gtccctttta 540
gaccctatgc agctcaggat aaagaagagc ttctgttatc catcagctca ttcattcatt 600
caaaaccatt tataggccag gtgcagtggc tcatgtttct aaccccagca ctttgggagg 660
ccaaggaagg aggatcactt gagcccagga gtttgagacc agcctgggca acatcacaaa 720
accctgtctc tacaaaaaat aaatcagtcg ggcgtggtgg cacaatcctg taatcccagc 780
tacttgagag tctaaggcag ggggaccaag ctcaggaggt cgaggctgca gtgagctatg 840
atcacgccac tgcactccag cctgggcaaa aaagcaagat cccaactcaa aaacaaaaca 900
aaacaaaaca aaacatgtat taagttcctg ccagacactg ttgcaggcac tggggattca 960
gcagagcaag actgaccagg ccccagatgc cttggaactg acgtaccacg gtggggcgga 1020
ggctggggga gacagacaca caaaacaacc aacaaggtgc tttctctttt ttattttatt 1080
ttattttatt ttattttatt tttttgagac agggtcttgc tctgttgccc aggctggagt 1140
gcagtggtgg gatcagaact cactgtagcc ttgacctcct gggctcaagc aatccctcca 1200
cctaagcctc ctgagtagct gggactacag gcatgcatca ccatacctgg ataatttttg 1260
tagtttttgt aaagacaggg ttttgccatg ttacccaggc tggattcaaa ctcctgggct 1320
caagcaatct gcccacctca gcctcccaaa gtgctgggat tacaggtgtg agccaccatg 1380
ccaggcccaa acaggacact ttctttcttt cttttttttg gagacggagt ctcactctgt 1440
tgcccaggct ggagtgcggt ggcatgatct cggctcactg caacctccac ctcctgggtt 1500
caagcaattc tcctgcctca acctcccaag tatctgggat tgcatccagg tgtgcaccac 1560
catgcctggc taatttttgt attttaagta gagacagggt tttgtcatgt tagcccacct 1620
ggtctcgaac tcctggcctc aagtgatccg cccactgtac ccgcctttca gacaaccact 1680
cagggctcac aggacttgaa aagtcaggag cccgcagggt tggtctatcc accagcttca 1740
aagaccactc tctcgcccac acaggtcgcc catggttttg agaccccggg atggccggtc 1800
tgccgcatgg ccagcctccc aaagtgttgg gattacaggc ataagccacc gtgttctgcc 1860
ccaaacagga tactttctaa taatactggg tgctatatca aggacagaac actggacctc 1920
aggtaaaggg accacaactg ctagtgagaa ggaggaagcc agggagctgg gcagagggca 1980
ctgccagtgc aaaggccctg tgggagaaga gctggtgcgt ttgaggaata gggaggaggt 2040
cagtgtggct ggagtggagt gaggcagggt gagggggaat aaggagcaag ttcaggctgg 2100
gaggcttgga gagcatttat tcagcaggaa cttgtattcc agcaaatttg gcagagaact 2160
cagtccacac atccctactg ggcctagaga gtggtcatgt tcagatcctg aacatctagg 2220
tagggaaatt aatatccggt aaactactat acatacttca aaacccatca tttttgtcca 2280
gtgggagtta aaaagccaca gcattttccc taatcccatc ccaccaactt tcccagaagt 2340
ccacccagag gaggagatga ggttcattta gctcttttcc tggccctcag cacaggctat 2400
tattagatat tcagtggatt tggggactgg gcgaggtggc tcagtcctgt aatcccagca 2460
ctttgggagg ccaaggctgg cagatcactt gagcccagga gtttgagacc agcctaagca 2520
acatggtggg accccatctc tacaaaaaat acaaaaagtt agctgggcat ggtggtgcat 2580
gcatgtagtc ccaggtactg aggaggctga ggtgggagga tcagctgagc ctgggaaggt 2640
cgaggctgca gtaagccatg atcacactac tatactccag cctggggcca gagtgagacc 2700
ctgtctcaaa aaaatggatg ttggacagaa aggagagaag gaaacccaaa atgcctagcg 2760
cccttcctgc aggacagcca tctggttgtt gggaccaaca gggacagatt taggcttagt 2820
ccaagcaaga atggcccaaa tttccaaaat tgagagctgc tgccctaaat gaggtagtga 2880
gttcctggcc cctgggaggt gtgcaagctg agcccggaca caattcattg aggatgctgt 2940
aggggaggtg ataccttccc tgggaaaagg ttggttggac caggtgacca ccccccagcc 3000
ctgcctcaaa tccctcccag ccctgagagt ctgggcttct gccctggccc tgagcactga 3060
gcccaccacc atcccccctg cccctggctt ccagccatga ccggctgctt tgtccttgaa 3120
ggcaagacca gtatgaagag ctgaaggacg aggccacctc ctgcagcctc cacaggtcgg 3180
cccacaatgc cacgcatgcc acctacacct gccacatgga tgtattccac ttcatggccg 3240
acgacatttt cagtgtcaac atcacagacc agtctggcaa ctactcccag gagtgtggca 3300
gctttctcct ggctgagagc agtgagtatc ctgggacccc aggcttaggg tggcactcaa 3360
tcttagctca ccgcagccct gacctctctg ggctcaggtg atcctcccac ctcagcctcc 3420
cgtgtagctg ggacaacagg tgtacaccac tacacccagc taatttttgt atttttagta 3480
gagacgggga tcttgccatg ttgcccaggc cagtctcgaa ctcctgggct ccagcgatcc 3540
acttgccttg gcctcccaaa gtgctaggat tataggtagg ggctgggtcc cctactccct 3600
gccctctggg ccagctctcc tccttccatc agatctgacc tggcctatgt cctcactaca 3660
ccttagagtt ctgttttgag ttgcgctatt ttttactaaa gtgtgaaagg ttgataatag 3720
gtttgagagg tattgaagga gctaggctgg gacccgggac ttctgggttc gtgttccggc 3780
tcagctctgc ccatctgtgt gaccatggat gaggcatacc tacctctggg acagcttccc 3840
tatttgtaga gcaaatgtcc ttttaaccct gactttctgg gatcctgacc ccataatgag 3900
gcgaatggac gacactgagt tacagaaaaa aaaaaccctc ttgggttttt tccccttcat 3960
aagctccagg gaggcctgga atttccctcc aggctggagg tagagggggg tcccaggggc 4020
aggagggcct ggttggtggg agcagtttta tctgtgagtt cagaattagg gcaggtggca 4080
gggcgaggtg gctcatgcct gtaattccaa cactttggaa ggctgaggca ggaggattgc 4140
ttgaacctgg ggctttaaga ccaacctggg caatatagcg agacctcatc tcaaaacaca 4200
aaacaagaac cagggcaggg gccaggcact gtggctcacg cctctaatcc cagcactttg 4260
ggaagctgag gcaggtggat cgtttgaggt caggtgttcg agaccagcct gggcaacatg 4320
gtgaaacccc atctctacta aaaatacaaa aaaatttgcc gggcgtggtg gcacacgccc 4380
gtaatcccgg ctactcggga ggctgaggca ggagaattgt ttgaacctgg gagacagagg 4440
ttgcaatgag ccaagatcat gccaccgcac tccagcctgg gcgtcagagt gagactccgt 4500
ctcaaacaaa acaaaacaga actaggcagg gcatggaagg gcctgggagg tggctttggc 4560
atggtgtgtg tgtgtgtgtg tgtgtgtgta tgtgtgtacc caaagcggcc ccagaggcaa 4620
acacaggctg caaagggagc ctgccctcag gaaggccccg cccatgagct cgggagtttc 4680
tgccgctaga ttcccagcag aggggctggg gtataagagg tgtctgtgat tcacactctg 4740
ggggtccagg tttgcacctc tttgctgagt ggatgtcaca ccacagagcg tgagagtgaa 4800
gcttcacaca ctcccagaca catgcacact cacgcaaatt cccagggtca cagactcacg 4860
cacacacaga ctcatagatg cgccctcaaa ctcacagatg tctccccagc cccagcctgc 4920
atggcatgga cacagaggcc cacactgcct ctctctcttg ctctctctca ctctcataca 4980
cactcacaca tacatacaca cacacacaca cacacacgta cacaatacac acacactcac 5040
acatatactc tcacatcact catacatggg cacacacacg cacacactca cacatgcatt 5100
ctcacacacg tgtagactca cacatgcaca cacatataca ctttctcaca cgtgtgcaca 5160
cacgcacata tacacatact catacactct cacatcacac acgggcacac atatgcacac 5220
actcacacat gcattctcac acacgtgtac actcatgcac agacacacac attactcacg 5280
ggcacacatg cacacactca cacatgcaca gactcacaag gggaccctcc ttgccacttg 5340
tcttgcagcg tgaatccaag catttctctc tggcattccc tccctccctg ccaggatctg 5400
cagctagcct ggtcccagct ggggcaggaa gtgattttct aaccacctcc cgccgagggt 5460
cccttcctcc tcctgggccc tgggcctcgc tgggctgagc agcccctctt cacgaccccc 5520
aggctctgca aacatctccc gagtacccac ggggcagttc tcagtaccct ggtgtcttct 5580
gggactcatc ctcacccagc cacacaggga agccgagagc cggcctccac ctgccccggg 5640
gaagacaaaa gaggggttag aaacgatttc agtcaaaaac atcccattac tggaacagca 5700
acagcaacaa cccagtgttc accatggcat tctgtaaaat gtgcacctct gtcacctcgg 5760
ggcgtggtgg gagccctcag tgcgggtttc aggatcacag gcagccttag agctcctttc 5820
tggaggcctt gcaaggttca ttagttaatc tgacaaatat atatatatat atatatatat 5880
atagagagag agagagagag agagagagag agagcgagca agcgcgcgcc agggtgtagc 5940
tttgtcctcc agcctggggt gcagtggcac aaccatagct cattgcagcc tggaactcct 6000
gggctcaagc gatcctccca cttcggcctc ctgagtggtt gggactacag gtgcacatca 6060
ccatcctggc taatttttaa attttttgta gagctgggat cttgctgtgt tgtccaggct 6120
ggtcttgaac acctggcctc aagcaatcct cccacctcag cctcccaaag tgctgagatt 6180
tttggcatga gccaccttgc ccagcctaat ttgccaaata tttattgagg gcctactgtg 6240
ccccaggctg tgttctagga gctgtgggca cagactgaac ctggaggaca aaacctctgc 6300
cccaaggagc taagagccca gtgagggagg cacatagtaa acaaaatcac agggtgagag 6360
gtgcagagtg ttagatggtg gcagattgaa agtgaagaag gtggctgggt gtggtggctc 6420
acacctgtag tcccagcact ttgggaggtt gaggtgggtg gatcacttga ggtcagaaga 6480
tcaagaccag tctggccaac atggggaaac tacttctcta ctaaaaatac aaaaattagc 6540
cagtcatggt ggtgcgcacc tatggtccca gctacttggg aggctgaggc cgtgtaatcg 6600
cttgggcttg ggaggtggag actgcagtga gctgagatca cgccactgca ctccagcctg 6660
ggtgacagag attctgtctc aaaaaaaaaa aaaaaaaaaa gaaagaagag aaaaaagaaa 6720
gggagggagg gagggaagga gggagaaagg aaggaaggaa ggagaacaaa aaaagaaaaa 6780
gtggagaagg tgagggcaga ggcctgcaac tcctctctga cactaataga cacttgtcac 6840
tcagttcctc tgaagttact gccttctgct aaatagcttc atccctttga acttgaagcc 6900
ccatgatata ggaactgagc ccattttaca agagaggaat ttaaggccct taacatattg 6960
tgttctgctc aagttcatga aatgtccttc gaatggagca aaaacagaat gccggctttg 7020
accttcacct ggaggccgcc catgaaagct tgctgttatg gctcagagct cggctcttgc 7080
acttgccagg agacgctgaa agatgaggac catcagattg gcctcccagg gccactgtgg 7140
ggtgtcaaac tgcccagagc agggcagcca ggcaggctct gtgtaaatgg cagccctcac 7200
cattagcatt catgctaaga agagtcatca actgcttgaa gtagctgggg ttgaaagaaa 7260
atgataccat catttagaac agaaatggaa aatgaatgga tttgccagca cgttgctgag 7320
aacatttttg gatgggttgc taactttaaa acttaatata ggccaggtgc agtggcttgt 7380
tcttgcaatc ccaggatttt gggaggctga ggtgggagca tcacttgagg ccaggagttt 7440
gagaccagcc tgggcaaaga tagtgagacc ccatttctac aaaacagaca aataattagc 7500
caagtgtggt cgcatccacc tgtagtctca gctacttggg aggctgaagt gggaggatgg 7560
cttgagccca ggaggttgag gctgctagtg agctatgatc gcaccactgc actccagcct 7620
gggcaacaca gtgagatcct gtctctaggg ggaaaaaacc cttggagatt tcacatgata 7680
attccagggg tctcccgttt cccattctgc acctcatatt gaaacttgcc cttcagtgca 7740
tagaaagcag ggtttcagat ttcatgtgtg tgcgtgtgca tttgtgtgtg tgcatgctgt 7800
gcgtgtgcat gtgtgttgca tgtgtaggca tgcgtttggg tgtgcatgtg tgggcatgca 7860
tgtgtgtggg tgtgcatgtg cacatgtgtt tgtgtatgcg tgtgtgtagg catgtgcgtg 7920
tgtgtttccc tagaaaactt agaaaagctg accactctgt gctgctgggg ccattcactt 7980
ttaactcttt tatttattta tttttgttcg tttttgagat ggagtctcgc tctgtcgccc 8040
gggctggagt gcagtggcgc aatcttggct cactccaagc tctgcctccc gggttcacgc 8100
cattctcctg cctcagcctc ccgagtagat gggactacag gcgcccgcca ccacgcccgg 8160
ctaatttttg tatttttagt aaagatgggg tttcaccgtg ttagccagga tggtctcgat 8220
ctcctgacct cgtgacccac ctgccttggc cttccaaagt gctgggatta caggcatgag 8280
ccactgcacc cagccagact gttttatttc gtcacagtcc ctaaacctgc ttcactcctc 8340
tctgtgacct gcccgcccct gagtttgcaa tcctgatcag gtctaaatct cccaggtccc 8400
agatgaataa actgaagccc agagttggga gtactttgcc caaagtcact cagggttgtt 8460
ggtcttcatc tcatccatga tgggtcaggc atggaggcag gggtgggggc aggcaggact 8520
tcctcccatc accaaatcct tttcacctgc agcaaattct caccccattt tcttttcctg 8580
ggtttttcca ccaagtcaag ccggctcccc ctttcaacgt gactgtgacc ttctcaggac 8640
agtataatat ctcctggcgc tcagattacg aagaccctgc cttctacatg ctgaagggca 8700
agcttcagta tgagctgcag tacaggaacc ggggagaccc ctgggctgtg gtgaggaatg 8760
tggggatcag tgcagctttg tgggagggga ggggagatga cggagtgcaa aggcatctgg 8820
gtgggagaga cgggtgaaca acatcattca tggccaagaa cagagaccaa gggcacgagc 8880
actcccttac agcgggccct cctcctcccc tcacttctcc ccaaggtagg agacccctga 8940
ataatctgtc ctccccaaac tggtctcctc ggcggtcagc atgccaaaga gactgtccct 9000
tccccagact ccaccccagg tctagcatag tcacaataga agaagtttca gaaatggggc 9060
aacctggaag cttcccaatt gcttagaatg gccatagttg gagagttcca gaaatatcaa 9120
ttattaagaa ggtccagcca ggtgcggtgg ctcatgcctg taatcccagc actttggagg 9180
ccaaggcagg tggatctctt gaggtcagga gtttgagacc agccagggca acatggtgaa 9240
actctgtttc tactaaaaat acaaaaaatt agccaggcgt ggtgacatgc acctgtaatc 9300
cagctattct ggaggcttag gcaggagaat cacttgaatc caggaggcgg aggttgcagt 9360
gagctgggat cgcgctactg cactctagcc tgggcaacag agtgagactc gatctcaaaa 9420
aaaaaaaaaa ggtccagtgt tagagggccc attgtttcaa acatgaaggt gttgagtgat 9480
gtccttccag cccactattt agaacagtca caagtgatgt actgtctcta aaaaataaaa 9540
aatagtaaca atgatagaac agtcacagct gaagagttct agaattgccc atcctttagg 9600
atagtcccag ctgaagatgt ccatgagtgc catactttag aaagtcccca gtggaagact 9660
tccagaggta tccattgcta agatgtctgt agttagagtt ctagaaatgc ctgctgttgg 9720
gaatgtcagt atgtcaactc ttggaattct tcaacagttg aagacattct ccactgatgt 9780
actgagactt atatcagtag tacatcggtt gagaatgtcc tcaactgttg aagaattcca 9840
aaagtatgca ttgcctggaa tgctgccaga ccattaccac tcctgagata tgcccagtcc 9900
tgcagggaga gtccactgcc aggactgtga gtggctgaat tggatcaaag agaaatcaaa 9960
cagagcatcc tatcacatgc acgggccagg cagtgctggt gagtctgtgc tccccatgcc 10020
tgagtccagc tactcaggca taggcttgat tcttgtggga agagaagaga cactcagccc 10080
ctttgatgga agctgggaaa agaggtggct ctgctggagg caggggctgg cctggtttaa 10140
ccctgacctg gtgcatcctt tccttgtact ggcagagtcc gaggagaaag ctgatctcag 10200
tggactcaag aagtgtctcc ctcctccccc tggagttccg caaagactcg agctatgagc 10260
tgcaggtgcg ggcagggccc atgcctggct cctcctacca g 10301
<210> 6
<211> 94
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
tctagctcaa gaccacacag ctaaggagtg actccaggtc agatatcgaa ttccgaagtt 60
cctattctct agaaagtata ggaacttcag gtct 94
<210> 7
<211> 109
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
ctctagaaag tataggaact tcatcagtca ggtacataat ggtggatccg atatcggggt 60
agagagatgt agtggttctg agatgttcta cggggtgatc tctgccaat 109
<210> 8
<211> 2607
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 8
agatagacac cgaagcctcc gctggtggcc ctgtgtttca gtcgcacaca gctgtctgcc 60
cacttctcct gtggtgtgcc tcacggtcac ttgcttgtct gaccgcaagt ctgcccatcc 120
ctggggcagc caactggcct cagcccgtgc cccaggcgtg ccctgtctct gtctggctgc 180
cccagcccta ctgtcttcct ctgtgtaggc tctgcccaga tgcccggctg gtcctcagcc 240
tcaggactat ctcagcagtg actcccctga ttctggactt gcacctgact gaactcctgc 300
ccacctcaaa ccttcacctc ccaccaccac cactccgagt cccgctgtga ctcccacgcc 360
caggagacca cccaagtgcc ccagcctaaa gaatggcttt ctgagaaaga ccctgaagga 420
gtaggtctgg gacacagcat gccccggggc ccagtggctg ccttactcct gctgattctc 480
catggagctt ggagctgcct ggacctcgtc tgctacaccg attacctcca gacggtcatc 540
tgcatcctgg aaatgtggaa cctccacccc agcacgctca cccttacctg gcaagaccag 600
tatgaagagc tgaaggacga ggccacctcc tgcagcctcc acaggtcggc ccacaatgcc 660
acgcatgcca cctacacctg ccacatggat gtattccact tcatggccga cgacattttc 720
agtgtcaaca tcacagacca gtctggcaac tactcccagg agtgtggcag ctttctcctg 780
gctgagagca tcaagccggc tccccctttc aacgtgactg tgaccttctc aggacagtat 840
aatatctcct ggcgctcaga ttacgaagac cctgccttct acatgctgaa gggcaagctt 900
cagtatgagc tgcagtacag gaaccgggga gacccctggg ctgtgagtcc gaggagaaag 960
ctgatctcag tggactcaag aagtgtctcc ctcctccccc tggagttccg caaagactcg 1020
agctatgagc tgcaggtgcg ggcagggccc atgcctggct cctcctacca ggggacctgg 1080
agtgagtgga gtgaccccgt catctttcag acccaggctg gggagcccga ggcaggctgg 1140
gaccctcaca tgctgctgct cctggctgtc ttgatcattg tcctggtttt catgggtctg 1200
aagatccacc tgccttggag gctatggaaa aagatatggg caccagtgcc cacccctgag 1260
agtttcttcc agcccctgta cagggagcac agcgggaact tcaagaaatg ggttaatacc 1320
cctttcacgg cctccagcat agagttggtg ccacagagtt ccacaacaac atcagcctta 1380
catctgtcat tgtatccagc caaggagaag aagttcccgg ggctgccggg tctggaagag 1440
caactggagt gtgatggaat gtctgagcct ggtcactggt gcataatccc cttggcagct 1500
ggccaagcgg tctcagccta cagtgaggag agagaccggc catatggtct ggtgtccatt 1560
gacacagtga ctgtgggaga tgcagagggc ctgtgtgtct ggccctgtag ctgtgaggat 1620
gatggctatc cagccatgaa cctggatgct ggccgagagt ctggccctaa ttcagaggat 1680
ctgctcttgg tcacagaccc tgcttttctg tcttgcggct gtgtctcagg tagtggtctc 1740
aggcttggag gctccccagg cagcctactg gacaggttga ggctgtcatt tgcaaaggaa 1800
ggggactgga cagcagaccc aacctggaga actgggtccc caggaggggg ctctgagagt 1860
gaagcaggtt ccccccctgg tctggacatg gacacatttg acagtggctt tgcaggttca 1920
gactgtggca gccccgtgga gactgatgaa ggaccccctc gaagctatct ccgccagtgg 1980
gtggtcagga cccctccacc tgtggacagt ggagcccaga gcagctagca tataataacc 2040
agctatagtg agaagaggcc tctgagcctg gcatttacag tgtgaacatg taggggtgtg 2100
tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tcttgggttg tgtgttagca catccatgtt 2160
gggatttggt ctgttgctat gtattgtaat gctaaattct ctacccaaag ttctaggcct 2220
acgagtgaat tctcatgttt acaaacttgc tgtgtaaacc ttgttcctta atttaatacc 2280
attggttaaa taaaattggc tgcaaccaat tactggaggg attagaggta gggggctttt 2340
gagttacctg tttggagatg gagaaggaga gaggagagac caagaggaga aggaggaagg 2400
agaggagagg agaggagagg agaggagagg agaggagagg agaggagagg agaggagagg 2460
ctgccgtgag gggagaggga ccatgagcct gtggccagga gaaacagcaa gtatctgggg 2520
tacactggtg aggaggtggc caggccagca gttagaagag tagattaggg gtgacctcca 2580
gtatttgtca aagccaatta aaataac 2607
<210> 9
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 9
tgattaatgt ttgatgtggg agggc 25
<210> 10
<211> 18
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 10
ggcccggcta cttaccag 18
<210> 11
<211> 20
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 11
gctcgactag agcttgcgga 20
<210> 12
<211> 21
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 12
agatgggcaa agcctggaca a 21
<210> 13
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 13
tgtgcctatg actgttgcag gtgtt 25
<210> 14
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 14
cctcgctatc ctaacatgcc agctc 25
<210> 15
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 15
aatgaatagg gtgtgcaggg tgcat 25
<210> 16
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 16
gcagcatccc cacagagaga aacta 25
<210> 17
<211> 21
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 17
ggatcggcca ttgaacaaga t 21
<210> 18
<211> 22
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 18
cagaagaact cgtcaagaag gc 22
<210> 19
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 19
cttgtcctca gcctcacaca acctg 25
<210> 20
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 20
ggtgaccatt catgggaagc gtgta 25
<210> 21
<211> 25
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 21
gacaagcgtt agtaggcaca tatac 25
<210> 22
<211> 24
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 22
gctccaattt cccacaacat tagt 24
<210> 23
<211> 529
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Met Pro Arg Gly Pro Val Ala Ala Leu Leu Leu Leu Ile Leu His Gly
1 5 10 15
Ala Trp Ser Cys Leu Asp Leu Val Cys Tyr Thr Asp Tyr Leu Gln Thr
20 25 30
Val Ile Cys Ile Leu Glu Met Trp Asn Leu His Pro Ser Thr Leu Thr
35 40 45
Leu Thr Trp Gln Asp Gln Tyr Glu Glu Leu Lys Asp Glu Ala Thr Ser
50 55 60
Cys Ser Leu His Arg Ser Ala His Asn Ala Thr His Ala Thr Tyr Thr
65 70 75 80
Cys His Met Asp Val Phe His Phe Met Ala Asp Asp Ile Phe Ser Val
85 90 95
Asn Ile Thr Asp Gln Ser Gly Asn Tyr Ser Gln Glu Cys Gly Ser Phe
100 105 110
Leu Leu Ala Glu Ser Ile Lys Pro Ala Pro Pro Phe Asn Val Thr Val
115 120 125
Thr Phe Ser Gly Gln Tyr Asn Ile Ser Trp Arg Ser Asp Tyr Glu Asp
130 135 140
Pro Ala Phe Tyr Met Leu Lys Gly Lys Leu Gln Tyr Glu Leu Gln Tyr
145 150 155 160
Arg Asn Arg Gly Asp Pro Trp Ala Val Ser Pro Arg Arg Lys Leu Ile
165 170 175
Ser Val Asp Ser Arg Ser Val Ser Leu Leu Pro Leu Glu Phe Arg Lys
180 185 190
Asp Ser Ser Tyr Glu Leu Gln Val Arg Ala Gly Pro Met Pro Gly Ser
195 200 205
Ser Tyr Gln Gly Thr Trp Ser Glu Trp Ser Asp Pro Val Ile Phe Gln
210 215 220
Thr Gln Ala Gly Glu Pro Glu Ala Gly Trp Asp Pro His Met Leu Leu
225 230 235 240
Leu Leu Ala Val Leu Ile Ile Val Leu Val Phe Met Gly Leu Lys Ile
245 250 255
His Leu Pro Trp Arg Leu Trp Lys Lys Ile Trp Ala Pro Val Pro Thr
260 265 270
Pro Glu Ser Phe Phe Gln Pro Leu Tyr Arg Glu His Ser Gly Asn Phe
275 280 285
Lys Lys Trp Val Asn Thr Pro Phe Thr Ala Ser Ser Ile Glu Leu Val
290 295 300
Pro Gln Ser Ser Thr Thr Thr Ser Ala Leu His Leu Ser Leu Tyr Pro
305 310 315 320
Ala Lys Glu Lys Lys Phe Pro Gly Leu Pro Gly Leu Glu Glu Gln Leu
325 330 335
Glu Cys Asp Gly Met Ser Glu Pro Gly His Trp Cys Ile Ile Pro Leu
340 345 350
Ala Ala Gly Gln Ala Val Ser Ala Tyr Ser Glu Glu Arg Asp Arg Pro
355 360 365
Tyr Gly Leu Val Ser Ile Asp Thr Val Thr Val Gly Asp Ala Glu Gly
370 375 380
Leu Cys Val Trp Pro Cys Ser Cys Glu Asp Asp Gly Tyr Pro Ala Met
385 390 395 400
Asn Leu Asp Ala Gly Arg Glu Ser Gly Pro Asn Ser Glu Asp Leu Leu
405 410 415
Leu Val Thr Asp Pro Ala Phe Leu Ser Cys Gly Cys Val Ser Gly Ser
420 425 430
Gly Leu Arg Leu Gly Gly Ser Pro Gly Ser Leu Leu Asp Arg Leu Arg
435 440 445
Leu Ser Phe Ala Lys Glu Gly Asp Trp Thr Ala Asp Pro Thr Trp Arg
450 455 460
Thr Gly Ser Pro Gly Gly Gly Ser Glu Ser Glu Ala Gly Ser Pro Pro
465 470 475 480
Gly Leu Asp Met Asp Thr Phe Asp Ser Gly Phe Ala Gly Ser Asp Cys
485 490 495
Gly Ser Pro Val Glu Thr Asp Glu Gly Pro Pro Arg Ser Tyr Leu Arg
500 505 510
Gln Trp Val Val Arg Thr Pro Pro Pro Val Asp Ser Gly Ala Gln Ser
515 520 525
Ser
<210> 24
<211> 3707
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
acttgctaca ctgactacct ctggaccatc acctgtgtcc tggagacacg gagccccaac 60
cccagcatac tcagtctcac ctggtgagta gccacgcctc caggccctga ggagctgctc 120
agagaagatg acaggatgtg gaggggctgc ctactgagcc cagggctgag ctaacctgtg 180
gcatgctctg tgggaagctt agctacaagc aagggcccgc cccttccacc agcaatgtca 240
atcactggga ttatgagctg ggagccaggt ccccacctta cagatgacca agctcacagg 300
actcacttgg atcacacaaa tgggaggctg tagtctgact tgctttccct catagacctt 360
cctgggttgg tagccctggg atggacagtt cccagtgacc ttctaagcac tgagtaggca 420
ctgtgtcctc accatgaata atttttaaat tattgctacc atgttccagt cgaggatact 480
gaatttcagt gaggtttagc cactggccca gattctcata gccagtgtac tcacaatccc 540
acacgatgtt ttaggataca caccacttgt ccttcaggcg ttcacccact ttaaacaagg 600
tttttgccag atgtgctgta ggggctgagg atcaaaacaa gactcgatac tgcagggtgg 660
gggagtcagg gatgctcagc tgggttggcc agggcattca tgccagcagg cacagcacgt 720
gcaaaggtca cgggtgagca aacgccacga gtgtcagctc ttgcttccag tgccacctga 780
ctgtgacctg agcagagtga gccagagtga ggggaaatag aagacacaga tggtggtggt 840
tgtgcagcag ccgtctgggc tgcatggaga agggcagctg gggtcaggga gctttgaagg 900
cccctctggc tgctacactg agagtgttgg agttcagaga tagcggagga gggagcaata 960
ccttcagaca gccgttacag ggcttgaggc tggaggctcg cttggaggag attgttcagc 1020
aaggtttgaa ttacaagaaa ttcagtgaag aactaagtaa agtggcagat tggtcctgga 1080
gggtgggtgg tcacgttcag gaaggaagaa aatatacctc cgaggacttc tatatgtact 1140
ctaaaaccca tccttttttg cccagtgaga cttaaagagc caaagcattc ctcactgtcc 1200
catctgacca atgtccctgg aagatctctc agagcagaga actactaagc cttactcccc 1260
tctaagcaca gagctgtggc tgtgccgtgg tggagggtgg gaaggagcga aagccagccc 1320
ggggtttcct atagctcagc agtcccaaag ctgagagctg ctgccctggg caagatagtg 1380
agtgtctggt cctctgaagt acgcaagctg tgtctggctg gactcagagc atgctgggtg 1440
ccgtagggag gggactgtcc ctgtgagagg ggctgcttgg accatgaccc tgagccctac 1500
ctccagccct tctcagttca agctttgctt tactgtcctg ccccaggcac tgagctccac 1560
tgtccccagc cccatgactg tctactctgt ccccaaaggc aagatgaata tgaggaactt 1620
caggaccaag agaccttctg cagcctacac aggtctggcc acaacaccac acatatatgg 1680
tacacgtgcc atatgcgctt gtctcaattc ctgtccgatg aagttttcat tgtcaatgtg 1740
acggaccagt ctggcaacaa ctcccaagag tgtggcagct ttgtcctggc tgagagcagt 1800
aagtaccatg agcccgaagc agagaggacc taggaccttt ctctgacctc agagcaagct 1860
cttctcactc ggccaaaatc tactctcttt ggtgtgtcct tttgagatgc acagctcagg 1920
tgtgatgttt tgatacgaga tttgggctaa atggccatgg agagccttag aggcttggaa 1980
aggtgtaggg caaggactcc actcagccct ccatctctgc ctgaccctga acaaggttta 2040
ccttgtaaag accctttaaa catgacccct aggattttgc tcctagtatg agcatcagtc 2100
agctaacatg aaatccttgc agctagatga gaggggccta aagttaggaa ggccccagga 2160
tggacctggc tatagctgtc acggagccca ggctgggcaa aggagggatc tggggcaggc 2220
agtggctttg gcgcatggca gcatgtgtat ctatagatac atatacatgt atgagtatta 2280
tatgtttata tgtatgcatc tctgtgtggt atggggcccc ttctttagga gagaaatcga 2340
aagcaacaga atccagtagg tgccttgctt aggacgtttc tgcattaggc actaatatta 2400
aactctaaga aactcccatc agccatctgg agtcccagct tttgagggtt tgtttttctc 2460
tttccactaa gaaaggacag aggatcttgg tctgtatgct tgtccccagc tgggctgggc 2520
tgagcctgag tcctgactct gcctttaagc ctgggagtcc ttctccacta tccccagtcc 2580
taggtctctg tttcagtcac ctgcctgcag tgatgagatc tagtccaaac gagcaggggc 2640
tgtggccagg gagtgctttg ttccaagtca gccagcggcc tttagacact gtcgggaggg 2700
agggactctg ctgcctgcag agtcttaccc tgttccatct ttctgggtct tcctccaagt 2760
caaaccagct ccccccttga acgtgactgt ggccttctca ggacgctatg atatctcctg 2820
ggactcagct tatgacgaac cctccaacta cgtgctgagg ggcaagctac aatatgagct 2880
gcagtatcgg aacctcagag acccctatgc tgtggtgagg tggctgggct ttggggtggg 2940
agctacaggg gtatccaggc aggtcccagt gtagtggcac aaggaacagg gcttagcatt 3000
ccctccctgc atgcaaccct ctccttcctt ctccctgggt cgaagaaccc ctcagaacat 3060
ctccacctgg actacttcct tacagtccac ctcagttaca gtggggcgag tttgggaaca 3120
tcccacttac ttggaatgtg aaggctactt gagagtgcac agctggaagt ttccaggatg 3180
gcccactgtt tagacagagc gagaatggtc ctggttgaag atttccataa aggacatcat 3240
ctagaatgtt cccaaggaga catttatagc catccattgg caagatggct gtaactaggg 3300
acttcccaaa aggtctactg ttaatgtcct atgtcatttg agacagggag ttcaggggct 3360
cactgtctgt gaagtttcta gttaaggaca tatagaggtg tccatggttg ctcccaagac 3420
attgatggtt ggtggtcagg ctactgccac agagagggag ctctagtcag ggatgtgcgc 3480
tccctgtggg ggcctgcgtg gtgtgacccc agtggaagga ctctgtgctc tactgatgac 3540
ctgagtgact acagagtgtc ctggttcttc tgtcctggca gaggccggtg accaagctga 3600
tctcagtgga ctcaagaaac gtctctcttc tccctgaaga gttccacaaa gattctagct 3660
accagctgca ggtgcgggca gcgcctcagc caggcacttc attcagg 3707
Claims (10)
1.一种IL21R基因人源化改造的非人动物的构建方法,其特征在于,所述的非人动物的基因组中包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列,所述的构建方法包括用包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列替换至非人动物IL21R基因座上。
2.根据权利要求1所述的构建方法,其特征在于,所述的非人动物的基因组中包含SEQID NO:5所示核苷酸序列,所述的构建方法包括用包含SEQ ID NO:5所示核苷酸序列替换至非人动物IL21R基因座上。
3.根据权利要求1或2所述的构建方法,其特征在于,所述的替换至非人动物IL21R基因座为替换非人动物IL21R基因中编码SEQ ID NO:1第24至211位的核苷酸序列,或者替换SEQID NO:24所示序列相同的核苷酸序列。
4.根据权利要求1或2所述的构建方法,其特征在于,所述的非人动物体内表达人或人源化IL21R蛋白,所述的人源化IL21R蛋白包括人IL21R蛋白的胞外区,所述的人源化IL21R蛋白包含SEQ ID NO:23所示的氨基酸序列。
5.根据权利要求1或2所述的构建方法,其特征在于,使用靶向载体进行非人动物的构建,其中,所述的靶向载体包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列或包含SEQ ID NO:5所示核苷酸序列,所述的靶向载体还包含5’臂和/或3’臂,所述的5’臂的核苷酸序列如SEQ ID NO:3所示,所述的3’臂的核苷酸序列如SEQ ID NO:4所示。
6.根据权利要求1或2所述的构建方法,其特征在于,所述的非人动物为大鼠或小鼠。
7.一种靶向载体,其特征在于,所述的靶向载体包含编码SEQ ID NO:2第24至211位氨基酸的核苷酸序列或包含SEQ ID NO:5所示核苷酸序列,所述的靶向载体还包含5’臂和/或3’臂,所述的5’臂的核苷酸序列如SEQ ID NO:3所示,所述的3’臂的核苷酸序列如SEQ IDNO:4所示。
8.一种人源化IL21R蛋白,其特征在于,所述的人源化IL21R蛋白包含人IL21R蛋白的胞外区,所述的人源化IL21R蛋白包含SEQ ID NO:2第24至211位氨基酸或包含SEQ ID NO:23所示的氨基酸序列。
9.一种编码权利要求8所述的人源化IL21R蛋白的人源化IL21R基因,其特征在于,所述的人源化IL21R基因包含SEQ ID NO:5所示核苷酸序列。
10.权利要求1-6任一构建方法构建的非人动物、权利要求8所述的人源化IL21R蛋白、权利要求9所述的人源化IL21R基因在IL21R基因或蛋白相关研究中的应用,所述的应用包括:
A)涉及人类细胞的免疫过程的产品开发,制造或筛选人类抗体中的应用;
B)作为药理学、免疫学、微生物学和医学研究的模型系统中的应用;
C)涉及人类细胞的免疫过程的生产和利用动物实验疾病模型,用于病原学研究中的应用;
D)在体内研究人IL21R信号通路调节剂的筛选、药效检测、评估疗效、验证或评价;或者,
E)研究IL21R基因功能,研究人IL21R抗体,研究针对人IL21R靶位点的药物、药效,研究免疫相关疾病药物以及抗肿瘤或炎症药物方面的用途。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116083483A (zh) * | 2021-10-12 | 2023-05-09 | 百奥赛图江苏基因生物技术有限公司 | Il21基因人源化非人动物及其构建方法和应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111197058A (zh) * | 2018-11-20 | 2020-05-26 | 北京百奥赛图基因生物技术有限公司 | 人源化cd73基因动物模型的制备方法及应用 |
-
2021
- 2021-02-08 CN CN202110170978.2A patent/CN112501204B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111197058A (zh) * | 2018-11-20 | 2020-05-26 | 北京百奥赛图基因生物技术有限公司 | 人源化cd73基因动物模型的制备方法及应用 |
Non-Patent Citations (6)
Title |
---|
NCBI GENBANK: "Homo sapiens interleukin 21 receptor (IL21R), RefSeqGene on chromosome 16,NCBI Reference Sequence: NG_012222.1,55393bp DNA linear", 《NCBI GENBANK》 * |
NCBI GENBANK: "interleukin-21 receptor isoform X2 [Pongo abelii],NCBI Reference Sequence: XP_003778635.1,538aa linear", 《NCBI GENBANK》 * |
NCBI GENBANK: "interleukin-21 receptor isoform X4 [Mus musculus],NCBI Reference Sequence: XP_036009203.1,268aa linear", 《NCBI GENBANK》 * |
姜雪峰等: "IL21/ IL21R信号在TNBS诱导小鼠炎症性肠病病变形成中的作用研究", 《微生物学杂志》 * |
汤海涛等: "IL21及其受体对小鼠急性肠炎调节机制", 《国际免疫学杂志》 * |
高大蕊等: "白细胞介素21作为治疗性药物的研究进展", 《中国新药与临床杂志》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116083483A (zh) * | 2021-10-12 | 2023-05-09 | 百奥赛图江苏基因生物技术有限公司 | Il21基因人源化非人动物及其构建方法和应用 |
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