猪流行性腹泻病毒蛋白对Vero E6核蛋白B23.1的影响

Effect of PEDV N protein on B23.1 protein in Vero E6 cell during infection with the porcine epidemic diarrhea virus

为了解猪流行性腹泻病毒(PEDV)核蛋白N对Vero E6细胞核仁蛋白B23.1的影响,将本实验室构建的DsRed-N1/B23.1重组质粒转染Vero E6细胞,转染6h后接种PEDV,分别在转染后12、18h和24h三个时间点,进行病毒感染细胞的间接免疫试验,同时设未感染对照组,而后利用激光共聚焦显微镜分析感染和转染细胞中B23.1蛋白的变化。结果表明,PEDV N蛋白与Vero E6核蛋白B23.1有共定位特征,在PEDV感染Vero E6细胞12h后,B23.1蛋白开始发生变化,随着时间的延长逐渐碎裂、分散。这个结果提示,病毒可能通过破坏核仁的结构和核仁蛋白的功能,进而为病毒的复制提供便利。

To understand the effect of PEDV N protein on B23.1 protein in Vero cell during infection with the porcine epidemic diarrhea virus,Vero E6 cells were transfected with recombinant plasmids pDsRed2-N1/B23.1,and the changes of B23.1 protein were observed using confocal microscopy analysis at 12,18 h and 24 h post transfection.Results showed that the PEDV N protein and the B23.1 phosphoprotein had co-localization features in co-transfected cells and B23.1 protein started changing at 12 h after infected PEDV,even crush with time extension,but the control group only had light changes.These results strongly suggested that viruses interact with the nucleolus to usurp host-cell functions and recruit nucleolar proteins to facilitate virus replication.