title: Prokaryotic expression of f protein from PPRV and characterization of its polyclonal antibody.
abstract: The goal of this study was to evaluate the specificity of a polyclonal antibody against the F protein from Peste des petits ruminants virus (PPRV). A pET30a/F prokaryotic expression vector was successfully constructed and its recombinant protein was expressed. The result of Western blot analysis showed that the fusion protein pET30a/F possessed good immunoreactivity and the purified recombinant protein was then used as the antigen to raise anti-pET30a/F polyclonal antibody in rabbits. The polyclonal antibody titer against the recombinant F protein was confirmed by indirect ELISA, and the protein's specificity against pET30/F polyclonal antibody was confirmed by both Western blot and indirect immunofluorescence assay in transfected cells. In short, we obtained the high-level expression of recombinant F protein as well as high titers of rabbit polyclonal antibody specificity against F protein in pCAGGS/F transfected cells. This special polyclonal antibody offers a valuable and useful tool for further study of the pathogenesis of PPRV early infection and the structural and functional characterization of PPRV F protein.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, PR China.
authors.forename: Qiuxia
authors.initials: Q
authors.lastname: Wang
authors.forename: Yongxi
authors.initials: Y
authors.lastname: Dou
authors.forename: Xiangfang
authors.initials: X
authors.lastname: Yang
authors.forename: Xuelian
authors.initials: X
authors.lastname: Meng
authors.forename: Junjun
authors.initials: J
authors.lastname: Zhai
authors.forename: Xueliang
authors.initials: X
authors.lastname: Zhu
authors.forename: Xuenong
authors.initials: X
authors.lastname: Luo
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.forename: Xuepeng
authors.initials: X
authors.lastname: Cai
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D017931:DNA Primers / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D018107:Peste-des-petits-ruminants virus / Q000737:chemistry*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014760:Viral Fusion Proteins / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000914:Antibodies, Viral; D017931:DNA Primers; C096524:F protein, Peste-des-petits-ruminants virus; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins