应用酵母双杂交系统筛选人胃黏膜上皮组织SHIP2的相互作用蛋白  

Screening of interacting proteins of SHIP2 in human gastric mucosal epithelium by yeast two-hybrid system

作　　者：徐良 钱学艺 任林 邵玉玲 李允允 叶艳 XU Liang;QIAN Xueyi;REN Lin;SHAO Yulin;LI Yunyun;YE Yan(Department of Immunology,School of Basic Medicine,Anhui Medical University,Hefei 230032;Precision Medicine Center,Yijishan Hospital of Wannan Medical University,Wuhu 241001,China)

机构地区：[1]安徽医科大学基础医学院免疫学教研室,安徽合肥230032 [2]皖南医学院第一附属医院弋矶山医院精准医学中心,安徽芜湖241001

出　　处：《细胞与分子免疫学杂志》2019年第11期1000-1007,共8页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81972264)。

摘　　要：目的通过酵母双杂交系统筛选人胃黏膜上皮组织标准均一化cDNA文库,寻找与含Src同源蛋白2肌醇-5-磷酸酶2(SHIP2)相互作用的蛋白。方法利用酵母双杂交系统,以SHIP2的P1(SH2+5-Ptase)和P2(PRD+SAM)段作为诱饵蛋白,筛选出人胃黏膜上皮组织均一化cDNA文库中与SHIP2相互作用的蛋白,并通过免疫共沉淀法进行验证。结果挑选出39个阳性克隆,经测序比对分析,回复性杂交,免疫共沉淀试验验证,最终确定一个与SHIP2相互作用的蛋白抗增殖蛋白1(prohibitin1/PHB)。结论酵母双杂交系统筛选人胃黏膜上皮组织SHIP2的相互作用蛋白PHB。Objective To screen a standard homogenous cDNA library of human gastric mucosal epithelium with yeast two-hybrid system and find the proteins that interact with Src-homology 2-containing inositol 5-phosphatase 2(SHIP2).Methods Using the yeast two-hybrid system,P1(SH2+5-Ptase)and P2(PRD+SAM)segments of SHIP2 were used as bait proteins to screen the proteins that bind to SHIP2 from the homogenous cDNA library of human gastric mucosal epithelium.The selected interacting proteins of SHIP2 were verified by co-immunoprecipitation assay.Results A total of 39 positive clones were selected and sequenced for alignment analysis.It was verified that PHB interacted with SHIP2 by reductive hybridization and co-immunoprecipitation assays.Conclusion PHB,the interacting protein of SHIP2 was screened by yeast two-hybrid system from the homogenous cDNA library of human gastric mucosal epithelium.

关 键 词：酵母双杂交系统 胃癌 含Src同源蛋白2肌醇-5-磷酸酶2(SHIP2) 抗增殖蛋白1(prohibitin1/PHB) 

分 类 号：R735.2[医药卫生—肿瘤] R392.11[医药卫生—临床医学]