酵母双杂交筛选人乳腺cDNA文库中与Gankyrin相互作用的蛋白

目的：应用酵母双杂交技术,筛选与Gankyrin有相互作用的蛋白质。方法：应用酵母双杂交系统,以Gankyrin基因全长为诱饵,在人乳腺cDNA文库中筛选能与Gankyrin相互作用的蛋白质,并运用营养缺陷型培养基和X-α-Gal等实验提供的信息,筛除假阳性克隆。结果：以Gankyrin为诱饵筛选出了5个阳性克隆,对其中一个蛋白质RhoGDI2与Gankyrin进行了免疫共沉淀与GSTpull-down实验验证,证实了它们的相互作用。结论：Gankyrin能与RhoGDI2发生相互作用,在此基础上,可以开展Gankyrin调控肿瘤发生机制的研究。

Screening of Proteins Interacting with Gankyrin by Yeast Two-Hybrid in Human Breast cDNA Library

Objective： To find proteins interacting with Gankyrin by using yeast two-hybrid. Methods： Yeast two-hybrid was applied to screen human breast cDNA library, and Gankyrin as the bait, respectivley. SD nutrient medium and X-αGal were used to exclude the false positive. Results： Five proteins were found to interact with Gankyrin, one of which, RhoGDI2, was confirmed the interaction by Co-IP and GST pull-down. Conclusion： Gankyrin can interact with RhoGDI2, and researches of the mechanisms of Gankyrin as a regulator of tumorigenesis can be carried out based on this interaction.