MxA基因真核表达载体构建及体外抗HBV作用研究

Study on construction of expression vector of human MxA gene and effect of anti-HBV in vitro

目的克隆人类MxA基因,探索其细胞内直接抗HBV作用,以便寻求HBV及其他病毒感染的新的基因治疗靶点。方法以pHMG-MxA质粒载体为基础,构建人类MxA基因表达质粒,采用人HBV-DNA转染的2.2.15细胞株作为研究对象,以不同浓度的MxA基因转染进行干预培养,lamivudine(3TC)阳性对照,同时设立空白对照,观察对HBV表达和复制的影响。结果经过酶切鉴定证明重组MxA基因表达质粒pcDNA-MxA构建正确,转染2.2.15细胞后呈现出明显抑制HBV复制和表达作用,且与转染剂量有关;3TC组也见MxA的表达增高,HBV-DNA复制受到明显抑制。结论抗病毒基因MxA具有细胞内抗HBV的复制及抑制病毒抗原基因的表达作用。

Objective To clone the human MxA gene and explore the directory anti-HBV effect of MxA gene, and to develop a new gene therapy for HBV or other viruses. Methods The expression vector of human MxA gene was constructed according to the cloned vector of pHMG-MxA. 2.2. 15 cells were cultured and transfected by the recombinant expression vector. Lamivudine was used as the postive control and blank was set. HBV expression and replication were examined. Results The recombinant expression vector of MxA gene was constructed correctly, which was identified through digestion of restriction enzyme. After recombinant expression vector transfection, MxA expression increased markedly. Replication and exprssion of HBV gene were inhibited significantly in 2.2.15 cells, which were related to the concentrations of the vector. MxA expression also increased by lamivudine and replication of HBV gene was inhibited almost completely. Conclusion Antiviral gene MxA has a directly inhibitory effect on HBV in vitro, which can inhibit HBV replication and exprssion in 2.2.15 cells.