摘要
利用PCR定点突变技术 ,以野生型大肠杆菌二氢叶酸还原酶基因为模板 ,获取 3种突变型 (Cys85Ser,Cys15 1Ser,Cys85 15 1Ser)二氢叶酸还原酶 (DHFR)基因 .用限制性内切酶BamHⅠ与PstⅠ将 3种突变基因片段插入到克隆载体pUC18上 ,进行蓝白筛选 ,将筛选的克隆进行DNA序列测定 .
By using the Polymerase Chain Reaction(PCR) site mutation technique,The three mutants (Cys85Ser,Cys151Ser,Cys85/151Ser)DNA fragment of Escherichia coli dihydrofolate reductase (DHFR) gene was amplified.The fragments were inserted into plasmid pUC18 by the sites of two restriction enzyme BamHI and PstI,and the target genes were confirmed by DNA equencing.
出处
《首都师范大学学报(自然科学版)》
2005年第1期63-67,共5页
Journal of Capital Normal University:Natural Science Edition
基金
国家自然科学基金项目 (3 980 0 0 2 8)
北京市科技新星计划项目 (9612 8)