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目的构建乙型肝炎病毒(HBV)C 区启动子调控的人β干扰素(IFN-β)基因的真核表达载体.方法切除真核表达载体 pcDNA3.1(+)-DT_(390)-VEG_(exon7)内部的 CMV 启动子及 DT_(390)序列使之成为 p3.1V_7;从逆转录病毒载体 pMNSM-IFN-β质粒中游离人β干扰素基因;用 PCR 方法从 pGEM.7Z-HBV 质粒中扩增人乙型肝炎病毒(HBV)的 C区启动子序列;通过转换酶切位点,将 HBV C 区启动子和人β干扰素基因共同插入到 p3.1V_7中,构建成受 HBV C 区启动子控制的人β干扰素基因真核表达载体 p3.1 V_7-HBV.CP-IFN-β.结果构建完成携带人β干扰素基因的人乙型肝炎特异性真核表达载体.结论该载体为慢性病毒性肝炎的特异性干扰素基因治疗奠定了基础.
OBJECTIVE: To construct an eukaryotic expression vector of human interferon beta gene (IFN-β) regulated by hepatitis B virus (HBV) promoter.Methods The eukaryotic expression vector pcDNA3.1 (+) - DT_ (390) (exon7) and DT_ (390) sequences into p3.1V_7. The human interferon beta gene was isolated from the retroviral vector pMNSM-IFN-β plasmid. The pGEM.7Z-HBV plasmid In which the HBV C promoter and human interferon beta gene were inserted into p3.1V_7 together to construct HBV C promoter region by converting the restriction site of hepatitis B virus (HBV) Region promoter-controlled human interferon beta gene eukaryotic expression vector p3.1 V_7-HBV.CP-IFN-β.Results Construction of human hepatitis B-specific eukaryotic expression vector carrying human interferon beta gene Conclusion The The vector lays the foundation for the specific interferon gene therapy of chronic viral hepatitis.