用Touch-Up PCR克隆中国仓鼠存活蛋白基因及其序列分析  

Cloning and Sequence Analysis of Chinese Hamster Survivin Gene

作　　者：郭俊伟[1] 赵兴卉[1] 于颖群[1] 孔毅荣[1] 易绍琼[1] 张晓鹏[1] 陈薇[1] 

机构地区：[1]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)

出　　处：《生物技术通讯》2007年第2期209-212,共4页Letters in Biotechnology

基　　金：国家自然科学基金项目(30570951)

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。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.

关 键 词：存活蛋白 Touch-UpPCR BIR结构域 

分 类 号：Q78[生物学—分子生物学]