| APOBEC3G真核表达载体的构建及其对HBV复制表达的影响 |
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| 引用本文： | 王鲁文,楮小刚,刘艳红,曹廷华,龚作炯.APOBEC3G真核表达载体的构建及其对HBV复制表达的影响[J].武汉大学学报(医学版),2009,30(2). |
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| 作者姓名： | 王鲁文 楮小刚 刘艳红 曹廷华 龚作炯 |
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| 作者单位： | 武汉大学人民医院感染科/武汉大学病毒学国家重点实验室,湖北,武汉,430060 |
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| 基金项目： | 国家自然科学基金资助项目(编号:30671876) |
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| 摘 要： | 目的:构建APOBEC3G真核表达载体,在体外初步探讨APOBEC3G对HBV复制表达的影响。方法:应用RT-PCR法从人PBMC细胞中扩增APOBEC3G基因,分子克隆法构建表达APOBEC3G蛋白的真核表达载体pcDNA3.1-APOBEC3G,并脂质体法转染HepG22.2.15细胞。Western-blot鉴定细胞中APOBEC3G蛋白的表达。ELISA法检测细胞培养上清液中HBsAg和HBeAg水平,实时定量PCR检测HBVDNA和HBV mRNA水平。结果:经酶切鉴定及测序证实APOBEC3G真核表达载体被成功构建,APOBEC3G蛋白可在HepG22.2.15细胞中表达。转染APOBEC3G重组质粒的HepG22.2.15细胞的HBsAg、HBeAg、HBVDNA及HBV mRNA水平均明显低于未转染重组质粒的HepG22.2.15细胞。结论:APOBEC3G明显抑制了HepG22.2.15细胞中HBV的复制与表达,成功构建APOBEC3G真核表达载体,为深入研究APOBEC3G抗HBV的作用机制奠定实验基础。
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| 关 键 词： | APOBEC3G 基因构建 真核表达 乙型肝炎病毒 HepG22.2.15细胞 |
Construction of Eukaryotic Expressional Vectors of APOBEC3G and the Effects on the Replication and Expression of HBV |
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| WANG Luwen,CHU Xiaogang,LIU Yanhong,CAO Tinghua,GONG Zuojiong.Construction of Eukaryotic Expressional Vectors of APOBEC3G and the Effects on the Replication and Expression of HBV[J].Medical Journal of Wuhan University,2009,30(2). |
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| Authors: | WANG Luwen CHU Xiaogang LIU Yanhong CAO Tinghua GONG Zuojiong |
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| Institution: | WANG Luwen,CHU Xiaogang,LIU Yanhong,CAO Tinghua,GONG Zuojiong Dept.of Infectious Diseases,Renmin Hospital of Wuhan University & State Key Laboratory of Virology of Wuhan University,Wuhan 430060,China |
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| Abstract: | Objective: To construct eukaryotic expression vector containing APOBEC3G,and to study the effects of APOBEC3G on replication and expression of HBV in vitro.Methods: The APOBEC3G gene was extracted from PBMCs by RT-PCR.Eukaryotic expression vector pcDNA3.1-APOBEC3G was constructed by molecular cloning and transfected into HepG2 2.2.15 cells by lipofectamine.The expression of protein APOBEC3G was detected by Western-blot in HepG2 2.2.15 cells.The levels of HBsAg and HBeAg in the media of transfected HepG2 2.2... |
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| Keywords: | APOBEC3G Gene Construction Eukaryotic Expression Hepatitis B Virus HepG2 2 2 15 Cells |
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