刚地弓形虫微线体蛋白6和醛缩酶在虫体内的定位  

Localization of microneme protein 6 and aldolase in Toxoplasma gondii

作　　者：尹志奎[1] 郑斌[2] 姚志军[2] 刘世国[2] 任红斌[2] 许兵红[2] 张海珠[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》2013年第2期86-89,共4页Journal of Xinxiang Medical University

基　　金：河南省科技攻关计划项目(编号:112102310209);河南省教育厅自然科学研究项目(编号:2012B310019);新乡医学院博士科研启动基金(2010)

摘　　要：目的观察刚地弓形虫微线体蛋白6(MIC6)和醛缩酶在虫体内的定位。方法聚合酶链反应扩增微线体蛋白2羧基端(MIC2C)基因片段,构建MIC2C/pGEX-4T-1/BL21表达系统,异丙基-β-D-硫代半乳糖苷诱导表达谷胱苷肽巯基转移酶-MIC2C(GST-MIC2C)蛋白,用该蛋白免疫新西兰兔,制备GST-MIC2C多克隆抗体。刚地弓形虫速殖子涂片,分别用一抗、荧光二抗温育,荧光显微镜下观察。结果成功获得GST-MIC2C多克隆抗体。荧光显微镜下显示MIC6(红色荧光)和醛缩酶(绿色荧光)2种蛋白皆定位于弓形虫的顶端。结论刚地弓形虫MIC6与醛缩酶在虫体内存在共定位关系,为2种蛋白相互作用关系的确立提供了细胞定位学证据。Objective To observe the location of microneme protein 6 （MIC6） and aldolase in Toxoplasma gondii. Methods The gene fragment of carboxyl terminus of microneme protein 2（MIC2C） was amplificated by polymerase chain re- action（PCR） and the expression system MIC2C/pGEX-4T-1/BL21 was constructed. The expression of glutathione-S-transfera- ses-MIC2C（GST-MIC2C） protein was induced by isopropyl-13-D-thigalactopyranoside and then this protein was used to immu- nize New Zealand rabbits to prepare the GST-MIC2C polyclonal antibody. The Toxoplasma gondii were spotted onto slides,and incubated with antibody, immunofluorescence antibody respectively, then were observeded by fluorescent microscope. Results The polyclonal antibody GST-MIC2C was obtained. The fluorescence microscopy displayed that MIC6 （red fluorescence） and aldolase （green fluorescence） were positioned at the top of the Toxoplasma gondii. Conclusion MIC6 and aldolase are co-lo- calized in Toxoplasma gondii,which provide the cellular localization evidence for protein-protein interaction.

关 键 词：刚地弓形虫 微线体蛋白6 醛缩酶 定位 

分 类 号：R382.5[医药卫生—医学寄生虫学]