人谷胱甘肽硫转移酶M1 TV2基因真核表达载体的构建与鉴定

Construction and identification of eukaryotic vector pcDNA3.1 for human glutathione-S-transferase M1 TV2 gene

目的 :构建人谷胱甘肽硫转移酶M1TV2 (GSTM1TV2 )基因真核表达载体并进行序列分析。方法 :用RT PCR技术从人肺脏组织总RNA中分离扩增人GSTM1TV2基因的cDNA序列 ,将其插入到真核表达载体pcDNA3.1多克隆位点中 ,构建真核重组表达质粒pcDNA3.1 GSTM1TV2 ,并用PCR扩增、酶切分析及序列测定等方法对重组质粒进行鉴定。结果与结论 :对人GSTM1TV2测序结果同GenBank序列完全一致 ,基因登陆号为AY5 32 92 7,表明已成功构建了pcDNA3.1 GSTM1TV2真核重组表达质粒 ,为进一步进行真核细胞转染 ,研究毒物、药物代谢的机制提供了理论依据。

Aim: To construct an eukaryotic expression vector carrying human glutathione-S-transferase M1(transcript variant2,GST M1 TV2) gene and analyse its sequence. Methods: The GST M1 TV2 cDNA was amplified from human lung total RNAs by RT-PCR and was recombined with eukaryotic expression vector pcDNA3.1.The recombined plasmid pcDNA3.1-GST M1 TV2 was verified with PCR,restriction analysis, and sequencing determination.Results and Conclusion: Human GST M1 TV2 has been recombined with pcDNA3.1. Compared with Genbank,it is correct.The gene number is AY532927.The recombined plasmid pcDNA3.1-GST M1 TV2 may be helpful to research on transfection into eukaryotic cell and machanism to toxin and drug matabolism.