title: Screening of genes expressed in vivo after infection by Vibrio anguillarum M3.
abstract: OBJECTIVE
  Genes uniquely expressed in vivo may contribute to the overall pathogenicity of an organism and are likely to serve as potential targets for the development of new vaccine. This study aims to screen the genes expressed in vivo after Vibrio anguillarum infection by in vivo-induced antigen technology (IVIAT).
  
  
  RESULTS
  The convalescent-phase sera were obtained from turbot (Scophthalmus maximus) survived after infection by the virulent V. anguillarum M3. The pooled sera were thoroughly adsorbed with M3 cells and Escherichia coli BL21 (DE3) cells. A genomic expression library of M3 was constructed and screened for the identification of immunogenic proteins by colony immunoblot analysis with the adsorbed sera. After three rounds of screening, 19 putative in vivo-induced (ivi) genes were obtained. These ivi genes were catalogued into four functional groups: regulator/signalling, metabolism, biological process and hypothetical proteins. Three ivi genes were insertion-mutated, and the growth and 50% lethal dose (LD(50) ) of these mutants were evaluated.
  
  
  CONCLUSIONS
  The identification of ivi genes in V. anguillarum M3 sheds light on understanding the bacterial pathogenesis and provides novel targets for the development of new vaccines and diagnostic reagents.
  
  
  CONCLUSIONS
  To the best of our knowledge, this is the first report describing in vivo-expressed genes of V. anguillarum using IVIAT. The screened ivi genes in this study could be new virulent factors and targets for the development of vaccine, which may have implications for the development of diagnostic regents.
authors.affiliation: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.
authors.forename: Y-X
authors.initials: YX
authors.lastname: Zou
authors.forename: Z-L
authors.initials: ZL
authors.lastname: Mo
authors.forename: B
authors.initials: B
authors.lastname: Hao
authors.forename: X-H
authors.initials: XH
authors.lastname: Ye
authors.forename: D-S
authors.initials: DS
authors.lastname: Guo
authors.forename: P-J
authors.initials: PJ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D005393:Fish Diseases / Q000276:immunology / Q000382:microbiology*; D005412:Flatfishes / Q000382:microbiology*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D014733:Vibrio / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D014735:Vibrio Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*
chemical_list: D001426:Bacterial Proteins