小反刍兽疫病毒H和F蛋白在细胞融合中的作用

【目的】研究小反刍兽疫病毒囊膜糖蛋白(血凝素蛋白和融合蛋白)在病毒囊膜和宿主细胞膜融合过程中所发挥的作用。【方法】制备构建成功的小反刍兽疫病毒囊膜糖蛋白和病毒受体SLAM、Nectin4的真核表达质粒pCMV-HA-H、pCAGGS-Flag-F、pCMV-Myc-SLAM和pCMV-Myc-Nectin 4,将其组合转染至CHO-K1细胞,通过显微观察和间接免疫荧光技术分析小反刍兽疫病毒H和F蛋白在病毒融合过程中的功能。【结果】除空白对照组和重组质粒单独转染组细胞中没有发现合胞体外,其余组细胞中均出现了合胞体,而且F和H蛋白共转染组合胞体的数目明显较多;并在共表达H、F蛋白的细胞中观察到了蛋白分布极化的帽子现象。【结论】PPRV F蛋白是病毒囊膜和细胞膜融合的必需蛋白,但需要与PPRV H共同作用才能使病毒成功入侵靶细胞。

Fusion function of Peste des petits ruminants virus envelope proteins

Objective] To study the role of Peste des petits ruminants virus envelope glycoproteins hemagglutinin and fusion during the fusion process. Methods] The eukaryotic expression recombinant plasmids pCMV-HA-H, pCAGGS-Flag-F, pCMV-Myc-SLAM and pCMV-Myc-Nectin 4 sequenced correctly were transfected or co-transfected into CHO-K1 cells, and then the fusion role of PPRV H and F proteins was analyzed by microscopy and indirect immunofluorescence technique. Results] The formation of syncytium was observed in the co-transfected cells while it was not found in the single plasmid transfected cells and the negative control. The number of syncytia in cells co-expressing H and F proteins was significantly more than that of the other groups. The H and F proteins were co-redistributed to form the polar caps (co-capping) in the same position. Conclusion] PPRV F protein is an essential protein for fusion of the viral membrane with host cell membrane, but it must interact with PPRV H to make virus invasion successfully.