酵母双杂交筛选与甲状旁腺素1型受体相互作用的蛋白  

Screening of the Proteins Interacted with Parathyroid Hormone Type 1 Receptor by Yeast Two-Hybridization

作　　者：刘俊丽[1] 冯凯[1] 路浩军[1] 秦亚亚[1] 张建中[1] 

机构地区：[1]解放军306医院病理实验科,北京100101

出　　处：《生物技术通讯》2010年第2期188-191,共4页Letters in Biotechnology

基　　金：306医院院级课题(07QN10);总后司令部课题;卫生部课题

摘　　要：目的:采用Clontech公司第3套酵母双杂交系统,筛选与甲状旁腺素1型受体(PTH1R)相互作用的蛋白。方法:将全长PTH1R基因克隆到载体pGBKT7作为诱饵,用LiAc介导将诱饵蛋白质粒pGBKT7-PTH1R和人肾cDNA文库质粒转化到酵母菌AH109中,检测Mel1报告基因的表达,并进行相互作用的验证。结果:测序和相互作用验证结果表明SNAPIN在酵母系统中能特异地与PTH1R相互作用;根据对SNAPIN基因的功能分析,其可能与细胞内钙稳态的调控有关。结论:为进一步研究PTH1R促进骨肉瘤进展的机制提供了一定的线索。Objective：To screen the proteins which can interact with parathyroid hormone type 1 receptor（PTH1R） by yeast two-hybridization system.Methods：The full long PTH1R gene was subcloned into the vector of pGBKT7 to construct the bait plasmid pGBKT7-PTH1R.A cDNA library from human kidney recombinant plasmids was as the target genes.They were simultaneously transformed into yeast cell AH109 by LiAc-mediated transformation method.And then,the expression of reporter gene Mel1 was tested.Results：SNAPIN could interact specifically with PTH1R was confirmed by sequence and protein interaction test again.SNAPIN seemed to associate with the regulation of intracellular calcium homeostasis according to its functional analysis.Conclusion：The results provide clues for the mechanism of PTH1R in promoting the progress of osteosarcoma.

关 键 词：甲状旁腺素l型受体 酵母双杂交 相互作用 SNAPIN 

分 类 号：Q78[生物学—分子生物学]