title: Expression and characterization of recombinant VP19c protein and N-terminal from duck enteritis virus.
abstract: BACKGROUND
  Previous studies have indicated that the VP19c protein and its homology play similar roles in capsid assembly of all Alphaherpesvirus subfamily. However, there is no report on the VP19c protein of duck enteritis virus (DEV). In this study, we expressed the DEV VP19c protein and presented its antigenic properties. Moreover, we developed polyclonal antibody against the VP19c protein and characterized it.
  
  
  METHODS
  A recombinant VP19c (rVP19c) and N-terminal were expressed in Escherichia coli (E.coli) and purified by Ni²+-affinity chromatography. The antigenic properties of the recombinant protein were determined by Western blot and indirect enzyme-linked immunosorbent assay (ELISA). Furthermore, the polyclonal antibodies against the purified recombinant proteins were produced and the titer of polyclonal antibody was determined by ELISA analysis. Finally, the antibody was used to recognize the VP19c in the cells infected with DEV in the immunofluorescence assay.
  
  
  RESULTS
  The N-terminally His-tagged rVP19c and rVP19c(N) were produced as inclusion bodies in E. coli strain BL21 (DE3) with molecular weight of about 66 and 46 kDa. Then the proteins were purified to reach the level of homogeneity. Western blot and ELISA analysis that the rVP19c seems to be structurally and antigenically very similar to native VP19c and the N-terminus of VP19c may contain most antigenic linear-epitopes. Furthermore, ELISA analysis demonstrated that the titer of polyclonal antibody was approximately 1:12800, and in the immunofluorescence assay, the antibody was able to recognize the VP19c in the cells infected with DEV.
  
  
  CONCLUSIONS
  To our knowledge, this was the first report on basic properties of DEV VP19c protein. In the present study, we obtained a high-level expression of the recombinant VP19c protein as well as high titers of rabbit polyclonal antibody against to VP19c protein. The anti-rVP19c serum was able to detect the VP19c protein in DEV infected cells and the VP19c protein targeted to the nucleus as distinct punctate speckles. This specific polyclonal antibody provides a good tool for further studying structural and functional characterization of DEV VP19c.
authors.affiliation: Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University,Ya'an, Sichuan, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xiang
authors.forename: Shunchuan
authors.initials: S
authors.lastname: Zhang
authors.forename: Anchun
authors.initials: A
authors.lastname: Cheng
authors.forename: Mingshu
authors.initials: M
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Chang
authors.forename: Chanjuan
authors.initials: C
authors.lastname: Shen
authors.forename: Dekang
authors.initials: D
authors.lastname: Zhu
authors.forename: Renyong
authors.initials: R
authors.lastname: Jia
authors.forename: Qihui
authors.initials: Q
authors.lastname: Luo
authors.forename: Zhengli
authors.initials: Z
authors.lastname: Chen
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Chen
mesh_terms: D000818:Animals; D015153:Blotting, Western; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D002467:Cell Nucleus / Q000737:chemistry; D002846:Chromatography, Affinity; D004372:Ducks; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005347:Fibroblasts / Q000821:virology; D015870:Gene Expression; D025922:Mardivirus / Q000235:genetics* / Q000276:immunology*; D008970:Molecular Weight; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D036022:Capsid Proteins; D011994:Recombinant Proteins