拟南芥翻译延伸因子EF1A的亚细胞定位研究

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摘要:

探讨了拟南芥的翻译延伸因子EF1A的亚细胞定位.以Col野生型拟南芥的cDNA为模板,通过高保真KOD扩增酶获得2 363bp的拟南芥转录延伸因子EF1A基因片段,将其与GFP融合后共同连入p1300表达载体,利用农杆菌侵染法将p35S∶GFP和p35S∶EF1A∶∶GFP转入野生型拟南芥中,观察转基因植物细胞中GFP的表达情况.结果显示:p35S∶GFP转基因植物中,细胞核中GFP荧光强度显著高于细胞质中荧光强度,但是在p35S∶EF1A∶∶GFP转基因植物中,细胞核中荧光强度与细胞质中荧光强度无显著差异,说明在进行蛋白翻译时细胞质中的EF1A含量显著高于细胞核中,这也为翻译延伸因子EF1A的亚细胞定位提供了实验依据.

The subcellular localization of the translation elongation factor EF1A of Arabidopsis was investigated in this study. The 2 363 hp cDNA fragment of the Arabidopsis translation elongation factort EF1A was cloned by KOD reaction from Arabidopsis Columbia. With GFP fusion, the gene fragmentconstructed into the p1300 vector, Then the p35 S： GFP and p35S：EF1A ：： GFP were trans formated into Col ecotypes ArabidopsisbyAgrobaeterium-mediation. GFP expression in transgenic plants cell was observed and showed that 35 S：GFP transgenic plants exhibited a significantly high GFP fluoresence in the nucleus than in cytoplasm,but in p35S：EF1A ：： GFP transgenic plants, the fluorescence intensity in nucleus was similar to cytoplasm. These results suggested that during protein translation, EF1A content in cytoplasm is higher than in nucleus. Furthermore, the subcellular localization of EFIA also been revealed in this study.

作者:

陈燕王伟倩张红莉夏群芳孙恒吉李瑞沙郑帮周树敏张卫

机构地区:

上海大学生命科学学院上海市能源作物育种及应用重点实验室

出处:

《betway官方app 学报：自然科学版》 CAS 北大核心 2015年第1期106-109,115,共5页

基金:

上海市教委浦江人才计划(08PJ1405500) 国家自然科学基金面上项目(30870225)

关键词:

拟南芥翻译延伸因子 EF1A GFP

Arabidopsis thaliana translation elongation factor EF1A GFP

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