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百脉根离子通道蛋白Pollux与翻译延伸因子EF1A的相互作用
引用本文:代广中,袁兆栋,苟洪兰,方 庆,朱 辉,张忠明. 百脉根离子通道蛋白Pollux与翻译延伸因子EF1A的相互作用[J]. 华中农业大学学报, 2012, 31(1): 34-38
作者姓名:代广中  袁兆栋  苟洪兰  方 庆  朱 辉  张忠明
作者单位: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
基金项目: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)和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。

关 键 词:百脉根  离子通道蛋白  酵母双杂交  翻译延伸因子
收稿时间:2011-04-28

Interaction between ion channel protein Pollux and elongation factor 1A of translation in Lotus japonicus
DAI Guang-zhong,YUAN Zhao-dong,GOU Hong-lan,FANG Qing,ZHU Hui,ZHANG Zhong-ming. Interaction between ion channel protein Pollux and elongation factor 1A of translation in Lotus japonicus[J]. Journal of Huazhong Agricultural University, 2012, 31(1): 34-38
Authors:DAI Guang-zhong  YUAN Zhao-dong  GOU Hong-lan  FANG Qing  ZHU Hui  ZHANG Zhong-ming
Affiliation:State Key Laboratory of Agricultural Microbiology/College of Life Science & Technology, Huazhong Agricultural University,Wuhan 430070,China
Abstract: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.
Keywords:Lotus japonicus  ion channel protein  yeast two-hybrid  EF1A
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