应用酵母双杂交系统筛选MRP2胞内区相互作用蛋白

Screening the interaction proteins of MRP2 by yeast two-hybrid system

目的应用酵母双杂交技术研究与多药耐药蛋白MRP2发生相互作用的蛋白。方法应用酵母双杂交系统,以人MRP2胞内区C末端220个氨基酸为诱饵,筛选成人肝cDNA文库,寻找能与之相互作用的蛋白。利用生物信息学分析,一对一酵母回复性杂交等提供的信息,筛除假阳性。结果经过酵母双杂交共获得162个克隆,经过验证分析,最终确定3个无重复阳性克隆。结论获得的3个基因编码的蛋白可能揭示MRP2新的作用机制。

Objective To study interaction proteins of MRP2 by yeast two-hybrid system. Methods Using the 220 C-terminal amino acids of MRP2 as bait, a human liver cDNA library was screened and the proteins interacting with MRP2 were searched. The false clones were eliminated by bioinformatic methods and one to one yeast two-hybrid system. Results Three true positive clones from yeast clones were obtained after verification. Conclusion The encoding proteins we obtained may play important roles in finding the new function of MRP2.