酵母双杂交筛选Clathrin相互作用蛋白  

Screening of proteins interacting with Clathrin by yeast two-hybrid system

作　　者：吕学军[1] 陆卫忠[1] 张永娟[2] 郭亮[1] 李少莹[1] 李玉英[1] 钱桂生[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸病研究所,重庆400037 [2]广东医学院生理学教研室,东莞523808

出　　处：《中华肺部疾病杂志（电子版）》2014年第4期17-19,24,共4页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：国家自然基金资助课题(81070052)

摘　　要：目的应用酵母双杂交技术从小鼠肺cDNA文库中筛选与Clathrin相互作用蛋白,进一步阐明Clathrin在急性肺损伤和急性呼吸窘迫症(ALI/ARDS)发生时肺泡上皮极性损伤中的具体作用机制。方法首先构建酵母双杂交pSos-Clathrin诱饵载体,酶切鉴定,然后确定Clathrin诱饵蛋白无自激活特性并检测了Clathrin诱饵蛋白的表达;最后筛选小鼠肺cDNA文库并对筛选得到的阳性克隆进行回转验证,对回转验证结果为阳性的文库克隆质粒送检测序,分析克隆序列。结果酵母双杂交筛选小鼠肺cDNA文库得到4个与Clathrin相互作用蛋白,分别是:腺苷酸环化酶关联蛋白1,细丝蛋白α,DAP凋亡诱导蛋白激酶2和G蛋白耦联受体激酶6。结论 Clathrin参与细胞极性调节、炎症损伤和细胞凋亡等过程。Objective To screen the protein that interact with Clathrin. Methods Yeast two-hybrid technique was applied to screen a mouse lung eDNA library with pSos-Clathrin as bait plasmid. Results The bait plasmid was constructed successfully and co-transformed together with a mouse lung eDNA library into yeast cells. At last four proteins interacting with Clathrin were identified： adenylate eyclase-associated protein 1; filamin alpha; DAP kinase-related apoptotic kinase 2 and G protein-coupled receptor kinase 6. Conclusions Clathrin was involved in regulation of cell polarity, inflammation and apoptosis.

关 键 词：网格蛋白 急性肺损伤 细胞极性 酵母双杂交 

分 类 号：R563[医药卫生—呼吸系统]