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目的为探讨利用水泡性口炎病毒(vesicular stomatitis virus,VSV)假病毒粒子系统包装西尼罗病毒(West Nile virus,WNV)的E蛋白可行性,构建3种E蛋白基因的真核表达质粒。以期用于WNV感染的流行病学调查。方法将表达WNV囊膜蛋白E的质粒体外转染293T细胞,然后感染VSV△G*G,并对E蛋白表达质粒进行改造,分别在基因序列前加入信号肽,在终止密码子之前加入VSV△G蛋白的细胞质尾序列。结果包装出的VSV△G*-WNVE滴度没有明显升高。提示WNVE蛋白与VSV的囊膜匹配性不佳,VSV△G*-WNVE作为病毒中和抗原代替野毒进行抗体筛查还需要进一步提高转染和包装效率。结论VSV作为高效、安全的新型载体、活病毒疫苗载体和肿瘤基因治疗载体以及高危病毒的研究工具具有巨大的研究价值和应用潜力。
OBJECTIVE To investigate the feasibility of packaging E protein of West Nile virus (WNV) by vesicular stomatitis virus (VSV) pseudovirus particles and to construct eukaryotic expression plasmids of three E protein genes. In order to be used for epidemiological investigation of WNV infection. Methods 293T cells were transfected with 293T cells in vitro and then infected with VSV △ G * G. The E protein expression plasmids were transformed. The signal peptide was added before the gene sequence, and VSV △ was added before the stop codon Cytoplasmic tail sequence of G protein. As a result, the VSV △ G * -WNVE titer did not increase significantly. Suggesting that WNVE protein and VSV capsule poor matching, VSV △ G * -WNVE as a virus neutralizing antigen instead of wild virus antibody screening also need to further improve the transfection and packaging efficiency. Conclusion As a new efficient and safe vector, the research tools of live virus vaccine vector, tumor gene therapy vector and high-risk virus have great research value and potential application.