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人IL-12 cDNA的克隆
引用本文:郭亚军,卫立辛,王皓,卢洋,周倩,刘小萍,王华菁,刘彦君.人IL-12 cDNA的克隆[J].第二军医大学学报,1999,20(7):431-434.
作者姓名:郭亚军  卫立辛  王皓  卢洋  周倩  刘小萍  王华菁  刘彦君
作者单位:第二军医大学东方肝胆外科医院国际合作肿瘤研究所
摘    要:目的: 克隆人IL-12的cDNA。方法:利用RT-PCR从人NC 37淋巴母细胞中克隆出IL-12中两个亚基p35和p40的cDNA并进行序列测定。结果:从NC 37细胞中克隆的p35 cDNA序列(GenBank登录号为AF 101062)与先前登录在GenBank上的M65271 (NC 37细胞来源)和M 65291(RPMI 8866细胞来源,美国专利号:5648467)的p35 cDNA序列分别有2处和1处碱基不同:编码第44位氨基酸残基密码子的第3个碱基M 65291是C,而M 65271和本研究AF 101062的相应序列2是G;编码第244氨基酸残基密码子的第3个碱基,M 65271序列是C,而M 65291和AF 101062的相应序列是T;编码第247氨基酸残基密码子的第2个碱基M 65271序列是C,而M 65291和AF 101062的相应序列是T。此外,本研究克隆的p40 cDNA序列与登录在GenBank的p40 cDNA (M 38444)序列完全相同。结论:尽管本研究克隆的p35 cDNA序列与已知的两种p35 cDNA序列分别有2处和1处碱基不同,但其编码的氨基酸序列却与专利登记的p35 cDNA(M 65291)编码的氨基酸序列完全一致,表明克隆的IL-12基因所表达的蛋白质在理论上是符合目前药用标准的。

关 键 词:白介素12  聚合酶链式反应

Cloning of human interleukin 12 cDNA
Liu Yanjun,Wang Huajing,Liu Xiaoping,Zhou Qian,Lu Yang,Wang Hao,Wei Lixin,Guo Yajun.Cloning of human interleukin 12 cDNA[J].Academic Journal of Second Military Medical University,1999,20(7):431-434.
Authors:Liu Yanjun  Wang Huajing  Liu Xiaoping  Zhou Qian  Lu Yang  Wang Hao  Wei Lixin  Guo Yajun
Abstract:Objective:To clone human IL-12 cDNA. Methods: IL-12 cDNA (p35 and p40 cDNA) was cloned by RT-PCR from human lymphoblastoid B cell line NC 37, and their sequences were determined by automatic DNA sequencer. Results: There were 2 bases and one base different in p35 sequences of GenBank M 65271 and M 65291 (patent number: 5648467) respectively comparing to our p35 sequence (AF 101062). The third base of 44th code of M 65291 was C, but that of M 65271 or AF 101062 was G. The third base of the 244th code of M 65271 was C, but that of M 65291 or AF 101062 was T. The second base of 247th code was C, but that of M 65291 or AF 101062 was T. In addition, the sequence of our p40 cDNA was identical with that of GenBank (M 38444). Conclusion: Although there are 2 bases and one base different in p35 cDNA sequence of Genbank M 65271 and M 65291 respectively, comparing to AF 101062, the sequence of corresponding amino acids of AF 101062 is identical with that patented one. The recombinant IL-12 expressed by our p35 and p40 cDNA is acceptable in theory.
Keywords:cDNA
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