人谷胱甘肽硫转移酶A1真核表达系统的构建及序列分析  

Construction of an Eukaryotic Expression Vector pcDNA3-Glutathione-S-Transferase A1 Gene From Human Liver cDNA Library and Partial Sequence Determination

作　　者：陈萍萍[1] 吴逸明[2] 张朝武[1] 吴拥军[2] 

机构地区：[1]四川大学公共卫生学院,四川成都610041 [2]郑州大学公共卫生学院,河南郑州450052

出　　处：《环境与健康杂志》2003年第3期144-146,共3页Journal of Environment and Health

基　　金：河南省自然科学基金资助项目(0111021400)

摘　　要：目的构建人谷胱甘肽硫转移酶A1真核表达系统,为毒理学、遗传药理学的研究等提供材料。方法用RT-PCR技术从人肝脏组织总RNA中分离扩增人谷胱甘肽硫转移酶A1基因的cDNA序列,将其插入到真核表达载体pcDNA3多克隆位点中,构建重组表达质粒,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。结果人谷胱甘肽硫转移酶A1被正确地克隆到真核表达载体pcDNA3中,测序结果同Genbank序列比较,在152位点T→C,氨基酸由Met→Thr。结论经酶切鉴定和PCR扩增,证实人谷胱甘肽硫转移酶A1真核表达系统构建成功。Objective To construct an eukaryotic expression vector carrying human glutathione-S-transferase(GST)A1gene and to provide study materials for toxicology and pharmacogenetic.Methods The GST A1cDNA was amplified and separated from human liver total RNAs by RT-PCR approach and recombined with eukaryotic expression vector pcDNA3.The recombined plasmid pcDNA3-hGSTA1was verified using PCR,restriction analysis and sequencing determination.Results Human GST A1gene was recombined correctly with pcDNA3,compared with Genbank,in code152T→C,amino acid Met→Thr.Conclusion The eukaryotic expression vector pcDNA3-hGSTA1was constructed for research on toxicology and pharmacogenetics.

关 键 词：谷胱甘肽硫转移酶A1 RT-PCR 基因克隆 序列测定 

分 类 号：R114[医药卫生—卫生毒理学]