摘要
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.
AIM Construction of eukaryotic expression vector carrying IFN-β gene under control of human HBV promoter. METHODS Internal CMV promoter and DT_(390) gene were deleted from eukaryotic expression vector pcDNA3.1 (+)- DT_(390)-VEGF_(exon7) to construct p3.1V_7.Human IFN-β gene was released from a retroviral vector pMNSM-IFN-β. Polymerase chain reaction (PCR) was employed to amplify promoter of HBV C region.After their restriction sites were changed,the promoter of HBV C region and human IFN-β gene were co-inserted into p3.1V_7.Construction of hepatitis-specific eukaryotic expression vector p3.1V_7- HBV.CP-IFN-β was completed. RESULTS The eukaryotic expression vector carrying IFN- β gene under control of human HBV promoter was constructed. CONCLUSION The vector is of great significance for hepatitis.specific IFN gene therapy.
出处
《世界华人消化杂志》
CAS
2000年第5期520-523,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.39770834~~
关键词
IFN-β基因
乙型肝炎病毒
真核表达载体
治疗
interferon-beta
hepatitis B viruses
tissue-specific gene therapy
eukaryotic expression vector
