| 1.3倍乙型肝炎病毒全基因真核表达载体的构建及其在Bewo细胞中的表达 |
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| 引用本文： | 资捷,王前,郑磊,熊石龙,蔡贞.1.3倍乙型肝炎病毒全基因真核表达载体的构建及其在Bewo细胞中的表达[J].热带医学杂志,2010,10(2). |
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| 作者姓名： | 资捷 王前 郑磊 熊石龙 蔡贞 |
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| 作者单位： | 南方医科大学南方医院检验医学中心,广州,510515 |
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| 摘 要： | 目的 构建1.3倍乙型肝炎病毒(HBV)全基因的真核表达载体pcDNA3.1(+)-HBV1.3,并观察其在Bewo细胞中的表达.方法 以质粒pMD18T-HBV上的HBV DNA序列为模板,构建1.3倍HBV全基因序列,将其插入到真核表达载体pcDNA3.1(+),用酶切、PCR及测序鉴定,并把该载体转染Bewo细胞,以Western免疫印迹、微粒子酶免疫分析法(MEIA)和荧光定量PCR法分别检测胞内和上清中HBsAg、HBeAg蛋白的表达及HBV DNA水平.结果 通过酶切、PCR及测序鉴定,成功构建1.3倍HBV全基因表达载体,该载体可以在Bewo细胞系中表达和分泌HBsAg与HBeAg,并可检测到高水平的HBV DNA.结论 成功构建了1.3倍HBV全基因真核表达载体pcDNA3.1(+)-HBV1.3,为研究胞内HBV宫内感染奠定了基础.
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| 关 键 词： | 乙型肝炎病毒 1.3倍HBV基因组 真核表达载体 Bewo细胞 |
Construction of Eukaryotic Expression Plasmids Containing A 1.3-fold Overlength Genome HBV and Its Expression in Bewo Cells in Vitro |
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| ZI Jie,WANG Qian,ZHENG Lei,XIONG Shi-long,CAI Zhen.Construction of Eukaryotic Expression Plasmids Containing A 1.3-fold Overlength Genome HBV and Its Expression in Bewo Cells in Vitro[J].Journal Of Tropical Medicine,2010,10(2). |
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| Authors: | ZI Jie WANG Qian ZHENG Lei XIONG Shi-long CAI Zhen |
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| Institution: | ZI Jie,WANG Qian,ZHENG Lei,XIONG Shi-long,CAI Zhen (Laboratory Medicine Center,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China) |
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| Abstract: | Objective To construct a recombinant eukaryotic expression vector pCDNA3.1(+)-HBV1.3, and determine the expression of the recombinant protein in Bewo cells. Methods The 1.3-fold overlength genome HBV DNA sequence was constructed from a recombinant plasmid pMD18T-HBV HBV DNA sequence and cloned into the eukaryotic expression plasmids pCDNA3.1 (+). The recombinant vector was confirmed by restriction enzyme digestion, PCR and sequencing. The plasmid was then transfected into Bewo cells.The level of extracellul... |
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| Keywords: | hepatitis B virus 1 3-fold over length genome HBV eukaryotic expression vector Bewo cell |
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