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用Touch-Up PCR克隆中国仓鼠存活蛋白基因及其序列分析
引用本文:郭俊伟,赵兴卉,于颖群,孔毅荣,易绍琼,张晓鹏,陈薇.用Touch-Up PCR克隆中国仓鼠存活蛋白基因及其序列分析[J].生物技术通讯,2007,18(2):209-212.
作者姓名:郭俊伟  赵兴卉  于颖群  孔毅荣  易绍琼  张晓鹏  陈薇
作者单位:军事医学科学院,微生物流行病研究所,病原微生物生物安全国家重点实验室(2004DAV00214),北京,100071
摘    要:目的:通过Touch-UpPCR(上升PCR)的方法从中国仓鼠卵巢细胞中克隆存活蛋白基因,并对其进行序列分析。方法:分别从293、H22、CHO-S细胞中提取总RNA并反转录得cDNA,然后用Touch-UpPCR克隆存活蛋白基因。结果:获得了存活蛋白基因,全长423bp,序列测定及同源性分析表明其与GenBank中报道的人以及小鼠的存活蛋白基因高度同源,同源性分别为84.8%和88.2%;氨基酸序列分析表明它们的蛋白产物具有功能相同的BIR结构域。结论:从中国仓鼠卵巢细胞中首次克隆到存活蛋白基因。

关 键 词:存活蛋白  BIR结构域
文章编号:1009-0002(2007)02-0209-04
修稿时间:2006-11-09

Cloning and Sequence Analysis of Chinese Hamster Survivin Gene
GUO Jun-wei,ZHAO Xing-hui,YU Ying-qun,KONG Yi-rong,YI Shao-qiong,ZHANG Xiao-peng,CHEN Wei.Cloning and Sequence Analysis of Chinese Hamster Survivin Gene[J].Letters in Biotechnology,2007,18(2):209-212.
Authors:GUO Jun-wei  ZHAO Xing-hui  YU Ying-qun  KONG Yi-rong  YI Shao-qiong  ZHANG Xiao-peng  CHEN Wei
Institution:2004DAV00214
Abstract:Objective:To clone the survivin gene using Touch-Up PCR in Chinese hamster ovary cell and to analyse its sequence.Methods:The total RNA from 293,H22,CHO-S cells was extracted and the survivin gene was amplified by Touch-Up PCR.Results:DNA sequence analysis revealed that the survivin gene was 423 bp.Sequence analysis shows that it was homologous with genes from human and mouse,amino acid sequence analysis shows that all of the three proteins contained the same BIR domain.Conclusion:The survivin gene using Touch-Up PCR in Chinese hamster ovary cell line CHO-S has been cloned for the first time.
Keywords:Touch-Up PCR
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