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.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2004年第2期181-184,共4页
Progress In Biochemistry and Biophysics
关键词
点突变
血管内皮生长因子
重叠PCR
基因突变
site directed mutagenesis, vascular endothelial grow factor, overlap PCR
