利用双杂交系统筛选并鉴定与GPR30相互作用的蛋白  

SCREENING OF GPR30 INTERACTING PROTEINS BY TWO-HYBRID SYSTEM

作　　者：张蕾[1] 常宏[2] 徐东[1] 

机构地区：[1]首都医科大学宣武医院心脏中心,北京100053 [2]河北医科大学基础课教学部生物化学教研室,河北石家庄050091

出　　处：《河北医科大学学报》2013年第4期378-380,F0002,共4页Journal of Hebei Medical University

摘　　要：目的筛选并鉴定与GPR30相互作用的蛋白,为GPR30蛋白亚细胞定位和功能研究提供线索。方法应用酵母双杂交技术,构建诱饵表达载体Pgbk/GPR30,与人卵巢互补DNA(complementary DNA,cDNA)文库共转化酵母菌AH109,筛选阳性克隆并测序。利用哺乳动物双杂交系统验证GPR30与候选蛋白的相互作用。结果从人卵巢cDNA文库中筛选出4个与GPR30结合的蛋白基因,分别是小G蛋白Rab8a、Rab40a、NM23-H2和TACC3。其中,小G蛋白Rab8a、Rab40a与GPR30的相互作用在哺乳动物双杂交系统中得到验证。结论小G蛋白Rab8a和Rab40a可能参与了GPR30细胞内转运或与GPR30参与的信号转导有关。Objective To screen and identify the GPR30 interacting proteins and provide clues for GPR30 protein subcellular localization and cellular function. Methods The bait expression vector pGBK/GPR30 was constructed and co-transformed with human ovarian cDNA library into yeast AH109 cells for yeast two-hybrid screening. Candidate interacting proteins were confirmed by mammalian two- hybrid system. Results The genes coding for four proteins that can bind to GPR30 were selected from human ovarian cDNA library, small G proteins Rab8a, Ra40a, NM23-H2 and TACC3. And the interaction between GPR30 and small G proteins RabSa, Rab40a was confirmed by mammalian two-hybrid system. Conclusion Small G proteins Rab8a and Ra40a may be involved in the intracellular transport of GPR30 or GPR30 mediated signal transduction by direct interaction.

关 键 词：受体 雌激素 双杂交系统技术 卵巢 

分 类 号：R349.81[医药卫生—基础医学]