酵母双杂交技术筛选与Axin相互作用的蛋白质  

Screening of Axin-interacting Proteins by Yeast Two-hybrid System

作　　者：叶志云[1] 张婷婷[1] 贺颖[1] 陈巧玉[1] 刘琪[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2009年第3期410-413,共4页Journal of Xiamen University：Natural Science

基　　金：国家基础学科人才培养基金(J0630649);福建省自然科学基金(2008J0109)资助

摘　　要：肿瘤抑制因子Axin以构架蛋白的形式通过Wnt、JNK、p53、TGF-β、G蛋白等众多信号转导途径参与细胞生长、增殖、分化和凋亡等多种重要细胞命运的调控过程.为了发现Axin新的相互作用蛋白,进一步研究Axin的多功能性,作者通过酵母双杂交技术,将Axin1基因从SpeI单一酶切位点到3′末端的DNA序列构建到pGBKT7载体作为诱饵,对小鼠脑的cDNA文库进行筛选,最后获得28个阳性克隆.通过对这些克隆的测序和对所得序列的生物信息学分析,得到13个与Axin的C末端相互作用的蛋白质,这些蛋白主要参与胚胎发育,肿瘤形成,蛋白翻译后修饰等生物学过程的调节.这一结果为进一步研究Axin的新功能提供了很好的线索.The tumor suppressor Axin regulates cell growth, proliferation and axis formation by participating in many important signal transduction pathways including Wnt,JNK, p53, TGF-β and G-protein pathways as a scaffold protein. In this study, we carried out a yeast two-hybrid screen for proteins that interact with Axin. The DNA fragment from the Spe 1 site to 3′-end of mouse Azinl gene coding the C-terminal region of Axin was suhcloned into pGBKT7 vector （SpeI-CT-BD）, and the mouse brain eDNA library was screened by this bait. We obtained 28 positive clones, 13 of which carried in-frame protein-coding sequences. These new interaction partners mainly related to embryo development ,tumorigenesis ,and protein modification. These results provided an important insight for further studying new functions of Axin,a multifunctional scaffold protein.

关 键 词：Axin 酵母双杂交 相互作用 

分 类 号：Q291[生物学—细胞生物学]