【摘要】 将西尼罗河病毒(WNV)的主要免疫原性基因prME插入HIV-1慢病毒载体pHR中,构建了转移质粒pHR-WNV/prME。在脂质体介导下,将该质粒与p△8.2、pVSV/G共转染293T细胞,48h后收获细胞上清。将上清转导正常的293T细胞,通过报告基因荧光观察、间接免疫荧光检测。结果表明,利用HIV-1慢病毒载体系统包装得到了假型病毒,可以高效转导293T细胞,并实现了WNVprME基因在被转导细胞中的表达,从而为基于慢病毒载体的WNV新型疫苗的研制奠定基础。更多还原

【Abstract】 prME,the major antigenic gene of West Nile virus(WNV),was inserted into HIV-1 lentiviral vector pHR,and the constructed transfer plasmid pHR-WNV/prME was co-transfected with p△8.2 and pVSV/G into 293T cells by Lipofectin.At hour 48 post-cotransfection,the supernatant of 293T cells was harvested and transducted into normal 293T cells.Subsequently,the expression of target protein was detected by indirect immunofluorescence assay.The results showed that the pseudotype virus pHR-WNV prME was efficiently transducted into 293T cells,and WNV prME protein gene was successfully expressed,providing the experimental basis for the development of new WNV-vaccines in the future.更多还原