【摘要】 为了观察鸡贫血病毒VP3基因在人肺癌A549细胞中的凋亡诱导情况,试验用PCR方法扩增了鸡贫血病毒的VP3基因,并将其克隆于真核表达载体pcDNA3.1上,构建含VP3基因的重组体,在体外利用LipofectamineTM2000介导的基因转染法将pcDNA-VP3转入人肺癌A549细胞中,用RT-PCR法检测VP3基因在细胞中的表达状况,同时利用流式细胞术检测验证VP3基因诱导凋亡的方式。结果表明:试验获得了VP3基因正确插入的真核表达载体;转染后VP3基因在细胞中得到了表达;鸡贫血病毒的VP3基因确以凋亡的方式诱导了肺癌细胞A549凋亡。更多还原

【Abstract】 To observation of the apoptosis inducing effect of chicken anemia virus(CAV) VP3 gene in human lung cancer cell Line A549.The fundamental cloning method,inserting the VP3 gene of chicken anemia virus into the eukaryotic expression vector pcDNA3.1 was used.Then,the positive recombinant containing VP3 gene was transfected into human lung cancer cell line A549,using Lipofect AMINETM-mediated transfection in vitro.After that,RT-PCR was performed to detect the expression condition of VP3 gene in vivo.After testified by restriction enzyme and sequencing analysis,VP3 gene was correctly inserted into eukaryotic expression vector.The result of RT-PCR suggests that VP3 gene could be expressed in this cell line.At the same time,the result of flow cytometric assay suggests that VP3 gene really could induce cell death by apoptosis.更多还原