百脉根离子通道蛋白Pollux与翻译延伸因子EF1A的相互作用  

Interaction between ion channel protein Pollux and elongation factor 1A of translation in Lotus japonicus

作　　者：代广中[1] 袁兆栋 苟洪兰 方庆[1] 朱辉[1] 张忠明[1] 

机构地区：[1]华中农业大学生命科学技术学院/农业微生物学国家重点实验室,武汉430070

出　　处：《华中农业大学学报》2012年第1期34-38,共5页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金(30870186);国家重点基础研究发展计划"973"课题(2010CB126502);国家转基因重大专项课题(2009ZX08009-116B)

摘　　要：利用酵母双杂交技术,以百脉根离子通道蛋白Pollux胞内结构域为诱饵,从百脉根AD-cDNA文库中筛选到与Pollux相互作用的多个阳性克隆,经DNA序列测定及NCBI数据库BLAST分析,其中有6个阳性克隆编码翻译延伸因子EF1A。进一步在大肠杆菌中表达与纯化Pollux和EF1A融合蛋白,通过Pull-down及Western杂交证实Pollux胞内结构域与EF1A的C-末端相互作用。Using ion channel protein Pollux-C containing RCK domain as a bait, 14 yeast colonies were screened from Lotus japonicus AD-cDNA library with yeast two-hybrid technique. Six positive clones were found to encode elongation factor 1A （EF1A） of translation through DNA sequencing and BLAST against NCBI database. The fusion tag protein of Pollux-C and EF1A expressed in E. coli were purified. The intra-cellular domain of Pollux interacted with the C-terminus of EF1A was verified by pull-down and western blotting in vitro. The results might lay a foundation for further researches on the regulating mechanism and biological function of EF1A in symbiosis signaling in legume.

关 键 词：百脉根 离子通道蛋白 酵母双杂交 翻译延伸因子 

分 类 号：S154.381[农业科学—土壤学]