小麦谷胱甘肽S-转移酶基因的克隆及原核表达  

Cloning and Prokaryotic Expression of TaGST from Triticum aestivum L.

作　　者：张蕾[1] 于永昂[1,2] 杨天佑[1] ZHANG Lei;YANG Tianyou;YU Yongang(School of Life Science and Technology, Henan Institute of Science and Technology, Collaborative Innovation Center of Modern Biological Breeding of Henan Province, Xinxiang 453003 , China;College of Agronomy , Northwest A & F University , Yangling 712100 , China))

机构地区：[1]河南科技学院生命科技学院,现代生物育种河南省协同创新中心,河南新乡453003 [2]西北农林科技大学农学院,陕西杨凌712100

出　　处：《华北农学报》2016年第3期38-43,共6页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(1305047);河南省教育厅科学技术研究重点项目(12A180011);河南省高等学校青年骨干教师资助计划项目(2014GGJS-100)

摘　　要：为了进一步研究小麦谷胱甘肽-S-转移酶基因(TaGST)的功能,采用RT-PCR方法分离了小麦谷胱甘肽-S-转移酶基因(TaGST)的ORF全长c DNA,并进行了生物信息学分析。结果表明:小麦TaGST基因的ORF全长690 bp,编码229个氨基酸;TaGST蛋白分子质量为25.81 k Da,p I为5.29。系统进化分析表明,该基因编码蛋白与水稻OsGST蛋白的氨基酸同源性最高,与已知植物GST家族成员的氨基酸序列聚类分析将TaGST聚为Phi类GST。构建原核表达载体p ET32-TaGST,对TaGST基因进行原核表达,SDS-PAGE结果表明,其所表达蛋白与预期蛋白大小一致。为进一步研究该基因的特性和功能奠定了理论基础。To investigate the function of TaGST gene,RT-PCR was used to obtain TaGST gene open reading frame sequence from wheat and analyzed by bioinformatics method. The sequence analysis results showed that the ORF of TaGST gene had a length of 690 bp coding for 229 amino acid,and the relative molecular weight of TaGST protein was 25. 81 k Da and its theoretical isoelectric point was 5. 29. Homology analysis showed that the amino acid sequence of TaGST was highly homologous with Oryza sativa,and phylogenetic analysis of the relationship of the newly identified TaGST with some known plantGSTs grouped the TaGST into the class of PhiGSTs. The prokaryotic expression of TaGST gene was done after construction of its prokaryotic expression vector p ET32-TaGST,and the SDS-PAGE results displayed that the expressed protein was consistent with the anticipated size. The results were expected to lay a foundation for further studies on the properities and function of this gene.

关 键 词：小麦 谷胱甘肽过S-转移酶 基因克隆 原核表达 

分 类 号：Q78[生物学—分子生物学] S512.03[农业科学—作物学]