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携带IFN-β基因的丙型肝炎病毒微型基因组构建及抑制病毒复制的效应

杨敬 雷迎峰 尹文 吕欣 陈任安 安群星 姚敏 贾战生 徐志凯

杨敬, 雷迎峰, 尹文, 吕欣, 陈任安, 安群星, 姚敏, 贾战生, 徐志凯. 携带IFN-β基因的丙型肝炎病毒微型基因组构建及抑制病毒复制的效应[J]. 临床肝胆病杂志, 2011, 27(1): 57-60.
引用本文: 杨敬, 雷迎峰, 尹文, 吕欣, 陈任安, 安群星, 姚敏, 贾战生, 徐志凯. 携带IFN-β基因的丙型肝炎病毒微型基因组构建及抑制病毒复制的效应[J]. 临床肝胆病杂志, 2011, 27(1): 57-60.
Yang Jing, Lei YingFeng, Yin Wen, Lu: Xin, Chen RenAn, An QunXing, Yao Min, Jia ZhanSheng, Xu ZhiKai. Construction of HCV minigenome containning IFN-β and investigation of its inhibitory effect on viral replication[J]. J Clin Hepatol, 2011, 27(1): 57-60.
Citation: Yang Jing, Lei YingFeng, Yin Wen, Lu: Xin, Chen RenAn, An QunXing, Yao Min, Jia ZhanSheng, Xu ZhiKai. Construction of HCV minigenome containning IFN-β and investigation of its inhibitory effect on viral replication[J]. J Clin Hepatol, 2011, 27(1): 57-60.

携带IFN-β基因的丙型肝炎病毒微型基因组构建及抑制病毒复制的效应

详细信息
  • 中图分类号: R512.63

Construction of HCV minigenome containning IFN-β and investigation of its inhibitory effect on viral replication

  • 摘要:

    目的构建丙型肝炎病毒(HCV)反应性干扰素(IFN)-β的微型基因组及其抑制效应的初步评价。方法提取poly I:C刺激的淋巴细胞总RNA,RT-PCR扩增IFN-β基因,克隆入前期构建的HCV微型基因组pT7-5U△131-315rI3Urz;PCR并鉴定插入方向,将反向插入的质粒命名为pT7-5U△131-315rIFNI3Urz。将微型基因组5U△131-315rIFNI3URz克隆入pcDNA3.1载体,获得pCMV-5U△131-315rIFNI3URz。将体外转录的5U△131-315rIFNI3URz RNA转染Huh7.5BB7细胞,48 h后检测IFN-β的表达。将pCMV-5U△131-315rIFNI3URz转染Huh7.5 JFH-1细胞系,荧光实时定量PCR法测定细胞中HCV RNA。结果成功建立了Huh7.5JFH-1细胞系;含有IFN-β的微型基因组在复制子细胞和HCV感染细胞中可以特异表达IFN-β;携带IFN-β的HCV微型基因组可以降低细胞中病毒的RNA拷贝水平,具有一定的剂量依赖效应。结论反向插入IFN-β基因的HCV微型基因组进入HCV感染细胞内...

     

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  • 出版日期:  2011-01-20
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