应用酵母双杂交技术筛选与p53相互作用的蛋白质

Screening of Proteins Interacting with p53 by Yeast Two Hybrid System

应用酵母双杂交技术筛选与p53发生相互作用的蛋白质,有助于进一步阐明其在肿瘤发生、发展中发挥重要作用的调控机制。采用酵母双杂交技术,以p53C末端(62-393aa)作为诱饵筛选人乳腺cDNA文库,寻找能与之相互作用的蛋白质,并运用报告基因等实验提供的信息,经过重复验证排除假阳性以确定最后的阳性克隆。以p53C末端(62-393aa)作为诱饵最终筛选出了一个阳性克隆,经测序及生物信息学分析,这一个阳性克隆为:PIAS3(Protein inhibitor of activated stat3)。说明TPIAS3蛋白能与p53(62-393aa)发生特异性的相互作用。

To find proteins interacting with human p53, Yeast two-hybrid was applied to screen human breast cDNA library, fragment of p53 （62--393aa）,as the bait . PIAS3, protein inhibitor of activated STAT3 , was identified to interact with p53 （62--393aa） . It is concluded that PIAS3 can interact with p53 （62--393aa）. Their interaction may suggest a new mechanism to reflect the function of p53.