| 百脉根离子通道蛋白Pollux与翻译延伸因子EF1A的相互作用 |
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| 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. |
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| 作者姓名： | 代广中 袁兆栋 苟洪兰 方 庆 朱 辉 张忠明 |
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| 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 |
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| 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） |
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| 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。
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| 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： | 2011/4/28 0:00:00 |
Interaction between ion channel protein Pollux and elongation factor 1A of translation in Lotus japonicus |
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| 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. |
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| Authors: | DAI Guang-zhong YUAN Zhao-dong GOU Hong-lan FANG Qing ZHU Hui ZHANG Zhong-ming |
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| Institution: | State Key Laboratory of Agricultural Microbiology/College of Life Science & Technology, Huazhong Agricultural University,Wuhan 430070,China |
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| 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. |
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| Keywords: | Lotus japonicus ion channel protein yeast two-hybrid EF1A |
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