应用酵母双杂交筛选Rab39A相互作用的蛋白  

Screening Proteins that Interact with Rab39A via Yeast Two-hybrid System

作　　者：范荣[1] 马琳琳[1] 张明[1] 王世聪[1] 贺威[1] 孙礼祥[1] 王团老[1] 

机构地区：[1]厦门大学药学院,福建厦门361100

出　　处：《生物技术》2013年第4期51-55,共5页Biotechnology

基　　金：国家自然科学基金面上项目("新Rab蛋白对甘露糖-6-磷酸受体及其介导的溶酶体蛋白质运输路径的调控作用及机制研究";No.30971442;31071176);福建省自然科学基金项目("小GTP酶Rab蛋白在人类乳腺癌发生过程中的调控作用";2010J01234)资助

摘　　要：目的:应用酵母双杂交方法,寻找可能与Rab39A相互作用的蛋白。方法:采用醋酸锂转化法将诱饵蛋白Rab39A转入AH109酵母菌,Western blotting检测蛋白表达情况,检测诱饵载体有无毒性和自激活能力,筛选出的蛋白通过免疫荧光法进一步验证。结果:获得12个与Rab39A相互作用的候选蛋白,免疫荧光法证明Rab39A与ATRN、TIFA、ITGB1蛋白确实存在互作关系。结论:筛选出的蛋白主要与细胞生长、肿瘤细胞形成有关,为Rab39A功能的研究提供了重要线索。Objective：Using Yeast- two -hybrid method to inquire proteins interacting with Rab39A. Method：The bait plasmid pGBKT7 -Rab39A was transfected into AH109 yeast, checking proteins expression, checking the auto - activity and toxicity of recombined plas- mid,the interactions were further confirmed with immunofluorescence experiment. Result： 12 positive proteins possibly interacting withRab39A were obtained. The interactions of Rab39A with ATRN ,TIFA, ITGB1 were further confirmed by immunofluorescence. Conclusion： The screened proteins have relation with cell growth, cancer cell formation, providing important clues for further investigation of biological function of Rab39A.

关 键 词：Rab39A 酵母双杂交 蛋白质相互作用 自激活 共定位 

分 类 号：Q786[生物学—分子生物学]