| 人Prohibitin 2哺乳动物细胞株的转染表达与定位的初步研究 |
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| 引用本文： | 武标,刘晓颖,范礼斌,邢昕,朱恒锐,汪云飞. 人Prohibitin 2哺乳动物细胞株的转染表达与定位的初步研究[J]. 安徽医科大学学报, 2010, 45(1): 5-8. DOI: 10.3969/j.issn.1000-1492.2010.01.002 |
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| 作者姓名： | 武标 刘晓颖 范礼斌 邢昕 朱恒锐 汪云飞 |
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| 作者单位： | 安徽医科大学生物教研室,合肥,230032;复旦大学遗传工程国家重点实验室,上海,200433;广东溢多利生物科技股份有限公司,珠海,519060 |
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| 基金项目： | 安徽省高校青年教师科研资助计划，安徽医科大学博士科研基金 |
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| 摘 要： | 目的 构建带FLAG标签的人类野生phb2 (prohibitin 2)表达载体,将其转染至HEK 293T细胞检测其表达,转染至COS7细胞观察phb2蛋白在细胞内的定位.方法 以含有人phb2全长cDNA序列的质粒为模板,通过带有FLAG标签上游引物,用PCR方法扩增phb2全长序列,然后插入真核表达载体pCDNA3中构建pCDNA3-FLAG-phb2表达载体,将构建的质粒转染至HEK 293T细胞,提取总蛋白,进行Western blot检测;转染至COS7细胞,在荧光显微镜下观察其细胞定位.结果 成功构建了pCDNA3-FLAG-phb2表达载体,转染表明此表达载体在HEK 293T细胞中能够有效表达,在COS7细胞中phb2呈斑点状主要在胞质内分布,细胞核中未见到明显的表达.结论 实验结果为进一步了解phb2的功能提供了一定的基础.
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| 关 键 词： | 基因表达 转染 印迹法 蛋白质 |
Human prohibitin 2 was transfected for expression and location of the preliminary study in mammalian cell lines |
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| Wu Biao,Liu Xiaoying,Fan Libin,et al. Human prohibitin 2 was transfected for expression and location of the preliminary study in mammalian cell lines[J]. Acta Universitis Medicinalis Anhui, 2010, 45(1): 5-8. DOI: 10.3969/j.issn.1000-1492.2010.01.002 |
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| Authors: | Wu Biao Liu Xiaoying Fan Libin et al |
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| Affiliation: | Wu Biao,Liu Xiaoying,Fan Libin,et al(Dept of Biology,Anhui Medical University,Hefei 230032) |
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| Abstract: | Objective To construct with FLAG tag expression vector for human wild phb2 and express this vector in appropriate mammalian cell lines to investigate the expression and localization of the product of phb2 gene.Methods The full length cDNA fragment of phb2 from pLenti6/V5-D-TOPO-phb2-WT plasmid vector was used to amplify with PCR,by virtue of the upstream primer with FLAG tags,then the fragment inserted into pCDNA3 vector to construct expression vector pCDNA3-phb2.Its expression was detected in HEK 293T cell... |
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| Keywords: | gene expression transfection blotting western |
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