用改进的重叠PCR引入血管内皮生长因子基因突变  

Site-Directed Mutagenesis of Vascular Endothelial Growth Factor (VEGF) by An Improved Overlap PCR

作　　者：白向阳[1] 吕安国[1] 吴文芳[1] 孙静[1] 牛瑞芳[2] 

机构地区：[1]中国科学院沈阳应用生态研究所,沈阳110016 [2]天津医科大学肿瘤医院中心实验室,天津300060

出　　处：《生物化学与生物物理进展》2004年第2期181-184,共4页Progress In Biochemistry and Biophysics

摘　　要：血管内皮生长因子 (vascularendothelialgrowthfactor,VEGF)的PCR产物克隆于T载体上 ,经转化JM1 0 9感受态菌株后 ,随机挑取 8个白斑菌落 ,混合后制成混合模板 .采用 3条引物 ,做两轮重叠PCR反应 ,获得了VEGF的突变基因 ,经PCR鉴定 ,酶切鉴定和测序分析表明所得基因为目的产物 .实践证明这种突变方法简单快速 。The PCR production of vascular endothelial growth factor (VEGF) was subcloned into a T vector, and the recombined plasmids were then transformed into competent cell JM109. Eight white clones were selected randomly, and the recombined plasmids were extracted and mixed as template. Three selected primers were adapted and two rounds of PCR were performed to introduce mutations. The production of those were confirmed by enzyme digestion, PCR amplification and sequencing. The method is proved to be a simple,quick and easy way to introduce site directed mutagenesis, and make latter work easy to do.

关 键 词：点突变 血管内皮生长因子 重叠PCR 基因突变 

分 类 号：Q754[生物学—分子生物学]