酵母双杂交筛选类固醇激素合成急性调节蛋白的相互作用蛋白质  

Screening of the Proteins Interacting with Steroidogenic Acute Regulatory Protein via a Yeast Two-Hybrid System

作　　者：杨晓扬[1] 于淼[1] 王建[1] 唐刘君[1] 陈慧[1] 金超智[1] 张卜兮[1] 詹轶群[1] 王晓辉[1] 杨晓明[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所蛋白质组学国家重点实验室,北京100850

出　　处：《生物技术通讯》2012年第1期6-10,共5页Letters in Biotechnology

基　　金：国家自然科学基金(30371596);国家重点基础研究发展计划(2002CB513100)

摘　　要：目的:利用酵母双杂交技术筛选人肝cDNA文库中与类固醇激素合成急性调节蛋白(STAR)相互作用的蛋白质。方法:将全长STAR基因克隆到酵母表达载体pDBLeu中,形成诱饵;将构建好的诱饵质粒转化至AH109酵母感受态中,利用酵母双杂交技术筛选人肝cDNA文库中与其相互作用的蛋白质,并进行相互作用的回转验证。结果:构建了酵母诱饵蛋白表达质粒pDBLeu-STAR,并筛选到6个猎物,其中有5对相互作用回转验证阳性。结论:为进一步研究STAR的功能和作用机制提供了新的线索。Objective： To screen proteins interacting with steroidogenic acute regulatory protein（STAR） via a yeast two-hybrid system.Methods： The full length gene of STAR was cloned into the pDBLeu vector as the bait plasmid.The plasmid was transformed into yeast cell AH109.Then,yeast two-hybrid assay was used to screen the proteins that can interact with STAR from the human liver cDNA library.Yeast two-hybrid re-transformation assay was used to confirm the interactions.Results： The full length STAR gene was successfully cloned into pDBLeu vector.Six preys were obtained via a yeast two-hybrid screening,and five of them were confirmed by yeast two-hybrid re-transformation assay.Conclusion： The results provide new clues for further research of STAR.

关 键 词：酵母双杂交 蛋白质相互作用 类固醇激素合成急性调节蛋白 

分 类 号：Q52[生物学—生物化学] Q78