肖芸,王季石,孙等军,任志娟,张维,吴正宇,张旭,方琴
XIAO Yun 1; WANG Ji-shi 2; SUN Deng-jun 3; REN Zhi-juan 2; ZHANG Wei 2; WU Zheng-yu 3; ZHANG Xu 2; FANG Qin 2 (1. Department of Clinical Hematological Examinations;

• 1:贵阳医学院附院血液检验科
• 2:贵阳医学院附院血液科
• 3:贵阳医学院附院药剂科
• 4:贵阳医学院附院血液科

摘要(Abstract)：

目的 :构建原核表达载体pPROEXHTb G15 6AMGMT ,观察突变型G15 6AMGMT在大肠杆菌中的表达情况。方法 :通过基因重组技术将G15 6AMGMT亚克隆至pPROEXHTb原核表达载体 ,在大肠杆菌DH5α中经异丙基硫代 β D半乳糖苷 (IPTG)诱导表达。 结果 :经PCR及酶切分析证实 ,成功构建了pPROEXHTb G15 6AMGMT原核表达载体 ,经SDS 聚丙烯酰胺凝胶电泳 (SDS PAGE)显示重组克隆化基因pPROEXHTb G15 6AMGMT在大肠杆菌DH5α中有表达。结论 :G15 6AMGMT在大肠杆菌DH5α中有表达 ,为获取突变型MGMT工程蛋白奠定了基础。
Objective: To construct recombinant prokaryotic expression vector pPROEX HTb-G156A MGMT and to observe its expression in E.coli DH5α. Methods: The G156 MGMT cDNA was subcloned into pPROEX HTb. G156A MGMT protein was induced by IPTG to express in E.coli. Results: The recombinant vector, pPROEX HTb-G156A MGMT, was successfully constructed and confirmed by the restriction endonuclease and PCR analysis. G156A MGMT protein expressed successflly in E.coli. Conclusions: G156A MGMT expresses in E.coli under the induction of IPTG. The study provides a foundation to the research aimed at obtaining mutant protein of G156A MGMT.

关键词(KeyWords)： 六氧甲基鸟嘌呤-DNA-甲基转移酶;大肠杆菌;基因表达
O 6-methylguanine-DNA-methyltransferase; Escherichia coli; gene expression

Abstract:

Keywords:

基金项目(Foundation): 贵州省科委基金资助项目(D2 0 0 0 - 5)

作者(Authors): 肖芸,王季石,孙等军,任志娟,张维,吴正宇,张旭,方琴
XIAO Yun 1; WANG Ji-shi 2; SUN Deng-jun 3; REN Zhi-juan 2; ZHANG Wei 2; WU Zheng-yu 3; ZHANG Xu 2; FANG Qin 2 (1. Department of Clinical Hematological Examinations;

DOI: 10.19367/j.cnki.1000-2707.2004.02.003

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