弓形虫微线体蛋白6,7,8与宿主Spata3,Dkk2蛋白相互作用的研究  

Interaction between Toxoplasma gondii Microneme Proteins 6,7,8 and Spata3,Dkk2 Proteins

作　　者：刘卿[1,2] 李法财[2] 杨文彬[2] 刘国华[1] 赵权[3] 朱兴全[1,2] 

机构地区：[1]湖南农业大学动物医学院,长沙410128 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,兰州730046 [3]吉林农业大学动物科学技术学院,长春130118

出　　处：《经济动物学报》2017年第2期68-74,共7页Journal of Economic Animal

基　　金：国家自然科学基金重点项目(31230073)

摘　　要：为了研究弓形虫微线体蛋白6(MIC6)、7(MIC7)及8(MIC8)是否与宿主精子生成相关蛋白3(Spata3)和Dickkopf相关蛋白2(Dkk2)互作,首先从弓形虫RH虫株cDNA中扩增了弓形虫MIC6、MIC7及MIC8基因,并从小鼠cDNA中扩增了Spata3和Dkk2基因。然后分别构建pGADT7-MIC6、pGADT7-MIC7和pGADT7-MIC8捕获载体及pGBKT7-Spata3和pGBKT-Dkk2诱饵载体。利用Clon Tech公司的MatchmakerTM Gold Yeast Two-Hybrid系统,将诱饵载体分别转化Y2HGold酵母菌,将捕获载体分别转化Y187酵母菌,最后通过酵母点对点验证分析弓形虫MIC6、MIC7和MIC8蛋白是否与宿主Spata3和Dkk2蛋白互作。结果表明:在酵母双杂交系统里MIC7和MIC8蛋白分别能够与宿主Spata3和Dkk2蛋白互作,而MIC6不能够与宿主Spata3和Dkk2蛋白互作。The objectives of the present study are to investigate whether the Toxoplasma gondii mi- croneme protein 6 （MIC6） , MIC7 and MIC8 interact with host spermatogenesis-associated protein 3 （Spata3） and dickkopf-associated protein 2 （Dkk2） using the yeast two-hybrid system. The T. gon- dii MIC6, MIC7 and MIC8 gene sequences were amplified from cDNA of T. gondii RH strain, and Spata3 and Dkk2 gene sequences were amplified from cDNA of mouse. Then, the PCR products were used to construct pGADT7-MIC6, pGADT7-MIC7 and pGADT7-MIC8 prey vectors, and pG- BKT7-Spata3 and pGBKT7-Dkk2 bait vectors, respectively. Using the Matchmaker Gold Yeast Two- Hybrid ClonTech system, the bait vectors were transformed into Y2HGold yeast, and the prey vectors were transformed into Y187 yeast. Then, the protein-protein interactions between MIC6, MIC7 and MIC8 and host Spata3 and Dkk2 proteins were determined by yeast small-scale matings. The results showed that MIC7 and MIC8 proteins were able to interact with host Spata3 and Dkk2 proteins, while MIC6 was not.

关 键 词：弓形虫 微线体蛋白 蛋白互作 Spata3 Dkk2 

分 类 号：Q933[生物学—微生物学]