期刊论文

中文

湖南中医药大学学报

1674-070X

2007

27

1

33-35

10.3969/j.issn.1674-070X.2007.01.012

湖南省2004年青年骨干教师培养资助项目；

本校为第一机构

目的 将DNA聚合酶δ相互作用蛋白1(PDIP1)编码序列562位碱基T突变为G.方法 利用定点突变PCR对PDIP1编码序列进行定点突变,然后将PCR定点突变产物和pGEX-4T-2载体经EcoR I与Sal I酶切,再利用连接酶将它们定向连接起来形成pGEX-4T-2-PDIP1重组质粒,转化E.coli DH5α感受态,时所得重组质粒进行酶切和测序.结果 第1轮和第2轮PCR都得到了预期大小的扩增片段,重组质粒经酶切也得到预期大小的片段,测序结果表明已成功将PDIP1编码序列第562位碱基T突变为G.结论 利用PCR定点突变是基因点突变的一种简单实用的好方法.

Objective To transit T at 562 base in PDIP1 to G. Methods PCR point mutation was induced by mutating primer and gene vector of PDIP1, the product of PCR and vector pGEX-4T-2 were digested by EcoR Ⅰ and Sal Ⅰ, the pGEX-4T-2-PDIP1 was ligased by the objective segments and them. Then, the recombined plasmid was transformed into E.coli DH5α and the recombined plasmid from Ecoli DH5α was sequenced and digested. Results The expected segments were gained in the first and second round of PCR, the recombined plasmid was testified to be right by cutt...