2011Zhongguo yufang shouyi xuebaoRequires access

Subcellular localization analysis between porcine epidemic diarrhea virus nucleocapsid protein and fibrillarin.

Da Shi, Jianfei Chen, Shi HongYan, Zhibang Zhang, Li Feng

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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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What this paper is about

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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Available 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.

Key concepts: Vero cell, Porcine epidemic diarrhea virus, Biology, Fibrillarin, Transfection, Plasmid, Virology, Western blot

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Subcellular localization analysis between porcine epidemic diarrhea virus nucleocapsid protein and fibrillarin. — Research Paper | ScholarLens