Adr亚型乙型肝炎病毒转染细胞模型的构建
Chinese Journal of Public Health(2006)
Abstract
目的建立乙型肝炎病毒(HBV)复制状态的细胞模型。方法利用分子亚克隆技术,将9600bp的adr亚型HBV全基因克隆至pcDNA3的EcoRI和HindIII位点构建pcDNA3-3HBV,通过脂质体介导的方法转染HepG2细胞,G418筛选。结果成功构建了质粒pcDNA3-3HBV,稳定转染后培养上清,ELISA检测结果显示,HBsAg、HBeAg阳性,且PCR检测前S/S基因阳性。PCR证实转染细胞中有HBV cccDNA存在,RT-PCR证实HBV S基因mRNA的表达。结论重组质粒pcDNA3-3HBV能在HepG2细胞中表达、转录、复制。
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Key words
adr subtype,transfection,eukaryotic expression plasmid,gene expression,hepatitis B virus
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