| 健康人APOBEC3G基因的克隆及真核表达载体的构建与鉴定 |
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| 引用本文： | 甘宜敏,孔凡运,史震,汤仁仙,郑葵阳.健康人APOBEC3G基因的克隆及真核表达载体的构建与鉴定[J].徐州医学院学报,2010,30(4):219-222. |
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| 作者姓名： | 甘宜敏 孔凡运 史震 汤仁仙 郑葵阳 |
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| 作者单位： | 徐州医学院病原生物学教研室,江苏,徐州,221002 |
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| 基金项目： | 徐州市科委课题，徐州医学院课题 |
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| 摘 要： | 目的克隆人载脂蛋白B mRNA编辑酶催化多肽样3G（apolipoprotein B mRNA editing enzyme,catalyticpolypeptide 3G,APOBEC3G）基因cDNA,构建APOBEC3G基因的真核表达载体,并在体外进行表达和鉴定。方法采用RT-PCR技术从健康人外周血单个核细胞中克隆APOBEC3G基因,将该基因插入pcDNA3.1真核表达载体,构建pcDNA3.1-A3G真核表达质粒,并利用脂质体将重组质粒pcDNA3.1-A3G转染HepG2和HL7702细胞,经免疫细胞化学、Western blot等方法检测APOBEC3G真核表达情况。结果双酶切鉴定和测序结果表明,APOBEC3G真核表达载体构建成功。免疫细胞化学和Western blot结果均显示,转染pcDNA3.1-A3G的HepG2和HL7702细胞APOBEC3G蛋白均高表达,转染空质粒pcDNA3.1和未转染的HepG2和HL7702细胞APOBEC3G蛋白低表达或不表达。结论成功构建了人载脂蛋白B mRNA编辑酶催化多肽样3G真核表达载体pcDNA3.1-A3G,为进一步研究APOBEC3G抗HBV作用奠定实验基础。
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| 关 键 词： | APOBEC3G 克隆 真核表达载体 |
The cloning of healthy human APOBEC3G gene and the construction and identification of eukaryotic expression vectors |
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| GAN Yimin,KONG Fanyun,SHI Zhen,TANG Renxian,ZHENG Kuiyang.The cloning of healthy human APOBEC3G gene and the construction and identification of eukaryotic expression vectors[J].Acta Academiae Medicinae Xuzhou,2010,30(4):219-222. |
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| Authors: | GAN Yimin KONG Fanyun SHI Zhen TANG Renxian ZHENG Kuiyang |
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| Institution: | GAN Yimin,KONG Fanyun,SHI Zhen,TANG Renxian*,ZHENG Kuiyang(Department of Pathogen Biology,Xuzhou Medical College,Xuzhou,Jiangsu 221002,China) |
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| Abstract: | Objective To clone the cDNA of human apolipoprotein B mRNA editing enzyme,catalytic polypeptide-like 3G(APOBEC3G) gene and construct the eukaryotic expression vector for the in vitro expression and identification.Methods The coding region of APOBEC3G gene was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs) of healthy humans,and then the gene fragment was inserted into the recombinant eukaryotic expression vector pcDNA3.1 to construct the eukaryotic expression plasmid of pcDNA3.1-A3G,which... |
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| Keywords: | APOBEC3G |
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