title: [Co-localization analysis between porcine epidemic diarrhea virus nucleocapsid protein and nucleolar phosphoprotein B23.1].
abstract: OBJECTIVE
  To elucidate the co-localization characteristic between porcine epidemic diarrhea virus (PEDV) N protein and B23.1 phosphoprotein.
  
  
  METHODS
  Two pairs of primers used to amplify N gene and B23.1 gene were designed and synthesized according to CV777 N gene sequence (AF353511) and human nucleolar phosphoprotein B23.1 gene sequence (BC050628.1), respectively. The PEDV N gene and B23.1 gene were amplified by RT-PCR from PEDV strain CV777 and Vero E6 cells, respectively; then cloned into eukaryotic expression vector pAcGFP1-C1 and pDsRed2-N1, to generate the recombinant plasmids pAcGFP1-C1/N and pDsRed2-N1/B23.1, respectively. Vero E6 cells were transfected with plasmids pAcGFP1-C1/N and pDsRed2-N1/B23.1.
  
  
  RESULTS
  The fusion proteins successfully expressed in transfected Vero E6 cells by western blot analysis, and the PEDV N protein and the B23.1 phosphoprotein showed co-localization features in co-transfected cells through confocal microscopy analysis.
  
  
  CONCLUSIONS
  The results will help to identify the nucleolar localization signals in PEDV N protein and to elucidate the mechanism of N protein located in nucleus.
authors.affiliation: Department of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Maojie
authors.initials: M
authors.lastname: Lv
authors.forename: Jianfei
authors.initials: J
authors.lastname: Chen
authors.forename: Hongyan
authors.initials: H
authors.lastname: Shi
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Chen
authors.forename: Xiuping
authors.initials: X
authors.lastname: Fan
authors.forename: Shichuan
authors.initials: S
authors.lastname: Shen
authors.forename: Li
authors.initials: L
authors.lastname: Feng
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018352:Coronavirus Infections / Q000662:veterinary / Q000821:virology; D000086462:Coronavirus Nucleocapsid Proteins; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D000090243:Nucleophosmin; D053485:Porcine epidemic diarrhea virus / Q000235:genetics* / Q000472:pathogenicity; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D013553:Swine Diseases / Q000821:virology; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000086462:Coronavirus Nucleocapsid Proteins; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D000090243:Nucleophosmin