弓形虫MIC6羧基端与醛缩酶相互作用的鉴定  

Identification on interaction of C terminal of microneme protein 6 with aldolase in Toxoplasma gondii

作　　者：郑斌[1] 尹志奎[2] 詹希美[3] 

机构地区：[1]河南省新乡医学院寄生虫学教研室,新乡453003 [2]河南省新乡医学院药理学教研室,新乡453003 [3]中山大学中山医学院寄生虫学教研室,广州510089

出　　处：《中国人兽共患病学报》2013年第9期883-885,890,共4页Chinese Journal of Zoonoses

基　　金：河南省科技攻关计划项目(No.112102310209);河南省教育厅自然科学研究项目(No.2012B310019);新乡医学院博士科研启动基金(No.2010)~~

摘　　要：目的鉴定弓形虫MIC6羧基端和醛缩酶的相互作用及两种蛋白在虫体内的定位。方法分别用GST-MIC6C多克隆抗体和兔免疫前血清结合的sepharose与弓形虫裂解液进行免疫共沉淀实验,实验产物进行SDS-PAGE和Western blot分析。弓形虫速殖子涂片,免疫荧光定位法确定两种蛋白在虫体内的定位。结果与对照相比,GST-MIC6C多克隆抗体的免疫共沉淀产物中有蛋白条带可分别被相应的抗体识别。荧光显微镜下显示,MIC6(红色荧光)和醛缩酶(绿色荧光)两种蛋白皆定位于弓形虫的顶端。结论弓形虫MIC6羧基端与醛缩酶存在相互作用并在虫体内存在共定位关系。This study aims to identify the interaction of C terminal of microneme protein 6 （MIC6C） with aldolase in Toxoplasma gondii and to observe the proteins location in tachyzoites of T. gondii. Anti-GST-MIC6C or preimmune rabbit se ra were added to protein G coated sepharose beads, and then parasite lysate was added to the beads and incubated. Beads were washed, and the protein-coated beads were dissolved in protein loading buffer. Proteins were resolved by SDS-PAGE and trans {erred to NC membranes for immunoblotting. Protein bands were found in co sediment products coming from immunoprecipita- tion （IP） experiment o{ anti-GST-MIC6C antibody, compared with the control （preimmune rabbit sera）. MIC6 and aldolase, examining by immunofluorescence （IF） microscopy of Toxoplasma tachyzoites smears, were apically localized. It＇s suggested that the MIC6C interacts with aldolase, and co-localizes in T. gondii.

关 键 词：刚地弓形虫 微线体蛋白6羧基端 醛缩酶 免疫共沉淀 共定位 

分 类 号：R382.5[医药卫生—医学寄生虫学]