细菌双杂交筛选大豆GmWNK1互作蛋白系统的建立及应用  

Establishment and Application of Bacterial Two-Hybrid System for Screening the Targets of GmWNK1 in Soybean

作　　者：王应祥[1] 郑焱[1] 梁翠月[1] 王秀荣[1] 庄楚雄[1] 严小龙[1] 廖红[1] 

机构地区：[1]华南农业大学根系生物学研究中心,广州510642

出　　处：《植物生理学通讯》2009年第4期372-378,共7页Plant Physiology Communications

基　　金：国家科技部重大基础研究项目(2005CB120902);国家自然科学基金(30230220);中国博士后科学基金

摘　　要：本研究利用大肠杆菌双杂交系统构建了一个高质量的大豆根系cDNA文库,同时利用大肠杆菌双杂交表达载体pBT构建了融合表达质粒pBT-GmWNK1,经酶切和测序鉴定、诱饵融合蛋白的表达检测及诱饵融合蛋白的自激活鉴定后作为诱饵,从大豆根部cDNA文库中筛选与GmWNK1发生互作的蛋白质,共获得18个阳性克隆。经测序和同源性比对发现,有10个阳性克隆编码已知蛋白,8个为假阳性。研究结果为揭示WNK基因家族的生物学功能和调控机制提供了重要的参考数据和研究材料。A soybean root cDNA library was constructed using the bacterial two-hybrid system. A bait plasmid pBT-GmWNK1 was also constructed and sequenced. After expression detection and self-activation identifica- tion of pBT-GmWNK1 fusion protein in bacterial two-hybrid system, we screened GmWNK1 interacting pro- teins and isolated 18 positive clones. Among these 18 positive clones, 10 encoded known proteins and 8 were false positive. These results provided important data for us to further investigate the biological functions and regulation mechanism of with no lysine kinase （WNK） family.

关 键 词：大豆 CDNA文库 细菌双杂交 GmWNK1 

分 类 号：S565.1[农业科学—作物学]