Subcellular localization analysis between porcine epidemic diarrhea virus nucleocapsid protein and fibrillarin.
Da Shi, Jianfei Chen, Shi HongYan, Zhibang Zhang, Li Feng
Abstract
Da Shi, Jianfei Chen, Shi HongYan, Zhibang Zhang, Li Feng
Abstract
To identify the interaction of porcine epidemic diarrhea virus(PEDV) N protein and fibrillarin in Vero E6 cells,the fibrillarin gene were amplified by RT-PCR from Vero E6 cells with a pairs of primers synthesized according to human fibrillarin gene,and cloned into eukaryotic expression vector pDsRed2-N1 to generate the recombinant plasmids pDsRed-Fib.Vero E6 cells were transfected with plasmids pAcGFP-N and pDsRed2-Fib.Western blot results showed that the fusion proteins expressed in transfected Vero E6 cells.Furthermore,the PEDV N protein and the fibrillarin showed co-localization features in nucleolar of co-transfected Vero E6 cells through confocal microscopy observation.The results would be helpful to elucidate the intereaction of PEDV N protein and nucleolar protein.
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To identify the interaction of porcine epidemic diarrhea virus(PEDV) N protein and fibrillarin in Vero E6 cells,the fibrillarin gene were amplified by RT-PCR from Vero E6 cells with a pairs of primers synthesized according to human fibrillarin gene,and cloned into eukaryotic expression vector pDsRed2-N1 to generate the recombinant plasmids pDsRed-Fib.Vero E6 cells were transfected with plasmids pAcGFP-N and pDsRed2-Fib.Western blot results showed that the fusion proteins expressed in transfected Vero E6 cells.Furthermore,the PEDV N protein and the fibrillarin showed co-localization features in nucleolar of co-transfected Vero E6 cells through confocal microscopy observation.The results would be helpful to elucidate the intereaction of PEDV N protein and nucleolar protein.
Key concepts: Vero cell, Porcine epidemic diarrhea virus, Biology, Fibrillarin, Transfection, Plasmid, Virology, Western blot