| 人MGMT基因Cdna的克隆表达及表达蛋白修复功能的鉴定 |
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| 引用本文： | 朱玉文,刘建国,黎高翔.人MGMT基因Cdna的克隆表达及表达蛋白修复功能的鉴定[J].生物工程学报,2001,17(4):396-399. |
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| 作者姓名： | 朱玉文 刘建国 黎高翔 |
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| 作者单位： | 中国科学院微生物研究所,北京,100080 |
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| 基金项目： | 国家自然科学基金资助项目(39990570). |
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| 摘 要： | 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。
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| 关 键 词： | O6-甲基鸟嘌呤-DNA-甲基转移酶 烷化剂 基因克隆表达 DNA分子修复 |
| 文章编号： | 1000-3061(2001)04-0396-04 |
| 修稿时间： | 2000年11月30 |
Cloning and Expression of MGMT cDNA and Analysis of the DNA Repair Activity of the Recombinant Protein |
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| ZHU Yu-Wen\ LIU Jian-Guo\ LI Gao-Xiang.Cloning and Expression of MGMT cDNA and Analysis of the DNA Repair Activity of the Recombinant Protein[J].Chinese Journal of Biotechnology,2001,17(4):396-399. |
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| Authors: | ZHU Yu-Wen\ LIU Jian-Guo\ LI Gao-Xiang |
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| Institution: | Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China. |
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| Abstract: | 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. |
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| Keywords: | O6-methylguanine-DNA methyltransferase(MGMT) alkylating agents DNA repair gene cloning and expression |
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