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目的 研究HBV反义全基因真核表达载体在抗HBV基因治疗的可能性。方法 采用基因重组技术 ,将HBV全基因片段反向插入真核细胞表达载体pBK -CMV克隆位点中 ,通过FuGENETM6转染试剂 ,将重组体转染人肝癌细胞系SMMC -7721细胞。结果 经RT-PCR表明 ,重组体导入的人肝癌细胞内有HBV反义RNA转录表达 ,且重组体导入对细胞生长无明显影响。结论 HBV全基因真核细胞表达载体在SMMC -7721细胞中能转录表达反义RNA ,因而有可能利用反义技术和转基因方法进行HBV全基因反义RNA抗HBV研究。
Objective To investigate the possibility of anti-HBV gene therapy using eukaryotic expression vector of HBV antisense gene. Methods The gene fragment of HBV was inserted into the eukaryotic expression vector pBK-CMV cloning site by gene recombination technique. The recombinant plasmid was transfected into human hepatoma cell line SMMC-7721 by FuGENETM6 transfection reagent. Results The results of RT-PCR showed that HBV antisense RNA transcripts were expressed in the recombinant human hepatoma cells, and the recombinant plasmid did not affect the cell growth significantly. Conclusion The eukaryotic expression vector of HBV eukaryotic cells can transfect antisense RNA in SMMC-7721 cells. Therefore, it is possible to use antisense technology and transgenic method to carry out HBV anti-HBV anti-HBV research.