人MGMT基因cDNA的克隆表达及表达蛋白修复功能的鉴定

Cloning and Expression of MGMT cDNA and Analysis of the DNA Repair Activity of the Recombinant Protein

克隆了Hela细胞O6 甲基鸟嘌呤 DNA 甲基转移酶 (MGMT)基因的cDNA序列 ,该序列与国外发表的cDNA完全一致。将此cDNA插入原核表达载体pET 2 1a后转化大肠杆菌BL2 1(DE3)获得表达的重组菌株pET 2 1a MGMT E .coliBL2 1(DE3) ,经IPTG诱导后产生分子量为 2 4kD的蛋白质。烷化类诱变剂致死突变实验表明 ,MGMT蛋白的表达能修复受体菌DNA分子因烷化类诱变剂导致的突变。

A cDNA encoding human O6-methylguanine-DNA methyltranferase (MGMT) was cloned from Hela S3 cells and the sequence is identical with the published data.The MGMT cDNA was inserted into the expression vector pET-21a and transformed into \%E.coli\% BL21 (DE3).A 24 kD protein was expressed after IPTG induction.Essays using lethal dose of alkylating agents indicate that expression of MGMT protein can repair the DNA mutations of the recombinant bacteria produced by alkylating agents.