刚地弓形虫谷胱苷肽巯基转移酶-醛缩酶融合蛋白的制备及纯化  

Preparation and purification of glutathione-S-transferases-aldolase of Toxoplasma gondii

作　　者：尹志奎[1] 郑斌[2] 王东[2] 刘世国[2] 许兵红[2] 张海珠[2] 任红斌[2] 

机构地区：[1]新乡医学院药理学教研室,河南新乡453003 [2]新乡医学院寄生虫学教研室,河南新乡453003

出　　处：《新乡医学院学报》2012年第5期327-329,共3页Journal of Xinxiang Medical University

基　　金：河南省科技攻关计划项目(编号:112102310209);新乡医学院博士科研启动基金资助(2010年)

摘　　要：目的体外制备刚地弓形虫醛缩酶(aldolase)蛋白并纯化。方法以刚地弓形虫cDNA链为模板,聚合酶链反应扩增aldolase基因,克隆至质粒pGEX-4T-1上,转化至大肠杆菌BL21;异丙基-β-D-硫代半乳糖苷诱导表达,亲和层析纯化表达产物。结果获得了aldolase基因序列,构建了aldolase/pGEX-4T-1原核表达系统,表达并纯化了谷胱苷肽巯基转移酶-醛缩酶(GST-aldolase)融合蛋白。结论体外获得了纯化的GST-aldolase蛋白,为后续的aldolase功能研究奠定了基础。Objective To obtain and purify the aldolase protein of Toxoplasma gondii in vitro.Methods The aldolase gene was obtained from cDNA library by polymerase chain reaction amplification,and subcloned into pGEX-4T-1 to generate translational fusion with glutathione-S-transferases（GST）.The recombinant protein GST-aldolase was expressed as a fusion protein with a GST tag in E.coli upon isopropyl-β-D-thigalactopyranoside induction and then purified with affinity chromatography.Results The aldolase gene was obtained,and the recombinant plasmid aldolase/pGEX-4T-1 was constructed successfully and expressed as a fusion protein.Conclusion The protein GST-aldolase is expressed in vitro,which may provide the foundation for the further studies on the function of aldolase.

关 键 词：刚地弓形虫 醛缩酶 表达 纯化 

分 类 号：R382.5[医药卫生—医学寄生虫学]