| B19病毒XA株VP1独特区真核表达系统的建立及鉴定 |
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| 引用本文： | 迟明,迟海峰,许东亮,王楠,聂晓晶,杨晓蕾,吕香萍,张国成. B19病毒XA株VP1独特区真核表达系统的建立及鉴定[J]. 现代生物医学进展, 2010, 10(23) |
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| 作者姓名： | 迟明 迟海峰 许东亮 王楠 聂晓晶 杨晓蕾 吕香萍 张国成 |
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| 作者单位： | 第四军医大学西京医院儿科;总参谋部老干部服务管理局卫生处; |
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| 摘 要： | 目的:克隆B19病毒XA株VP1u基因,构建真核重组表达载体.方法:从已构建好的B19病毒XA株原核表达载体中获得VP1u基因,将其克隆入真核表达载体plRES2-EGFP中,经酶切鉴定并测序验证后,获得真核表达载体plRES2-EGFP-VP1u.将其转染至HeLa细胞,提取细胞总蛋白,用Western blot技术检测VP1u蛋白的表达.结果:成功构建了携带人B19病毒VP1u基因的真核表达载体plRES2-EGFP-VP1u,荧光显微镜下可见pIRES2-EGFP-VP1u转染HeLa细胞后表达EGFP蛋白而发出绿色荧光,Western blot证明VP1u蛋白在HeLa细胞中表达.结论:成功构建了携带人B19病毒VP1u基因的真核表达载体plRES2-EGFP-VP1u并在HeLa细胞中正确表达,为今后B19病毒VP1u基因疫苗的研究奠定基础.
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| 关 键 词： | VP1u基因 基因克隆 表达载体 |
Construction and Identification of a Eukaryotic Expression Plasmid Encoding VP1-unique Region of Parvovirus B 19-XA |
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| CHI Ming,CHI Hai-feng,XU Dong-liang,WANG Nan,NIE Xiao-jing,YANG Xiao-lei,LV Xiang-ping,ZHANG Guo-cheng. Construction and Identification of a Eukaryotic Expression Plasmid Encoding VP1-unique Region of Parvovirus B 19-XA[J]. Progress in Modern Biomedicine, 2010, 10(23) |
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| Authors: | CHI Ming CHI Hai-feng XU Dong-liang WANG Nan NIE Xiao-jing YANG Xiao-lei LV Xiang-ping ZHANG Guo-cheng |
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| Abstract: | Objective:To clone VP1-unique region of parvovirus B19-XA and to construct eukaryotic expression plasmid.Methods:VP1-unique region of parvovirus B19-XA was acquired from the prokaryotic expression plasmid pQE30-VP1u,and was cloned into vec-tor pIRES2-EGFP.The recombinant plasmids were identified by sequencing.The recombinant plasmid were transfected into HeLa cells.Then Western blot and fluorescence microscopy were used to identify the expression of VP1u and green fluorescence protein.Results:The VP1u gene ... |
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| Keywords: | VP1u gene Gene cloning expression vector |
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