| Adr亚型乙型肝炎病毒转染细胞模型的构建 |
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| 引用本文： | 赵艳芳,闫永平,张磊,王安辉,苏海霞,门可,张景霞,徐德忠.Adr亚型乙型肝炎病毒转染细胞模型的构建[J].中国公共卫生,2006,22(9):1066-1068. |
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| 作者姓名： | 赵艳芳 闫永平 张磊 王安辉 苏海霞 门可 张景霞 徐德忠 |
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| 作者单位： | 第四军医大学军事预防医学系流行病学教研室,西安,710032 |
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| 摘 要： | 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HBVcccDNA存在，RT—PCR证实HBVS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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| 关 键 词： | 乙型肝炎病毒 adr亚型 真核表达质粒 转染 基因表达 |
| 文章编号： | 1001-0580(2006)09-1066-03 |
| 收稿时间： | 12 16 2005 12:00AM |
| 修稿时间： | 2005年12月16 |
Construction of cell models transfected with HBV adr subtype |
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| ZHAO Yan fang , YAN Yongping , ZHANG Lei ,et al..Construction of cell models transfected with HBV adr subtype[J].Chinese Journal of Public Health,2006,22(9):1066-1068. |
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| Authors: | ZHAO Yan fang YAN Yongping ZHANG Lei |
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| Affiliation: | ZHAO Yan fang , YAN Yongping , ZHANG Lei , et al . |
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| Abstract: | Objective To establish HBV replication cell models. Methods Using molecular subclone technique,pcDNA3-3HBV was constructed by insertion of 9?600?bp fragment of HBV adr subtype genome into the EcoRI and HindIII sites of pcDNA3.HepG2 was transfected with pcDNA3-3HBV by using lipofectamine transfection reagent and screened with antibiotic G418. Results The plasmid pcDNA3-3HBV was constructed successfully.After stable transfection,HBsAg and HBeAg were positive in the supernatant which tested by ELISA.S gene and pre-S gene were also positive which tested by PCR.cccDNA was found in the cells by PCR,and HBV pre-S1 mRNA was measured by RT-PCR. Conclusion pcDNA3-3HBV can be expressed,transcribed and replicated in HepG2 cells. |
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| Keywords: | hepatitis B virus adr subtype eukaryotic expression plasmid transfection gene expression |
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