cDNA文库基础上运用热启动聚合酶链反应末端延伸快速分离全长cDNA序列  

Rapid Isolation of Full-Length cDNA Sequence from cDNA Library by Hot-Start Polymerase Chain Reaction

作　　者：闾宏伟[1] 杨向东[2] 何淑雅[1] 杨永宗[2] 

机构地区：[1]南华大学生物化学与分子生物学教研室,湖南省衡阳市421001 [2]南华大学心血管病研究所

出　　处：《中国动脉硬化杂志》2002年第5期396-399,共4页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金 (3 0 2 0 0 10 3 );国家重大基础研究项目 (G2 0 0 0 0 5 690 5 );湖南省自然科学基金 (0 0JJY2 0 2 7);湖南省教育厅课题 (0 0C15 1)资助

摘　　要：建立一种cDNA文库基础上运用热启动聚合酶链反应末端快速扩增分离人类新基因全长cDNA序列的新技术。以文库载体序列与待扩增目的基因片段为模板各设计引物进行热启动聚合酶链反应扩增 ,从人胎肝cD NA文库中获得氧化型低密度脂蛋白诱导U937细胞泡沫化过程中差异表达序列标签片段 (FRG4 )的全长cDNA序列 ,聚合酶链反应产物克隆到pGEM T载体中 ,并测序鉴定 ,从而成功获得FRG4的全长cDNA序列。Aim To establish a new technique for rapid isolation full length cDNA sequence of human novel gene from cDNA library using hot start PCR. Methods The library vector specific primers and gene specific primers have been designed for performing hot start PCR to attain the full length cDNA of differential expressed sequence tag(FRG4) of U937 foam cell formation induced by ox LDL from human fetal liver cDNA library. The products of PCR are cloned to pGEM T vector and sequenced. Results We have successfully attained the full length cDNA sequence of FRG4. Conclusion This technique is a rapid and efficient method for isolating full length cDNA sequence of human novel gene from cDNA library.

关 键 词：CDNA 聚合酶链反应 热启动 序列标签 

分 类 号：R362[医药卫生—病理学]