title: Production of specific IgY antibody to the recombinant FanC protein produced inEscherichia coli.
abstract: OBJECTIVE
  Enterotoxigenic Escherichia coli (ETEC) strains are one of the primary causes of diarrhea in newborn calves and in humans, pigs, and sheep. IgY technology has been identified as a promising alternative to generating a mass amount of specific antibody for use in immunotherapy and immunodiagnostics. The purpose of this study was to produce specific antibody by egg yolk antibody (IgY) to recombinant FanC protein from ETEC.
  
  
  METHODS
  FanC (K99) gene was amplified from ETEC by specific primers and polymerase chain reaction. The gene was cloned and subcloned into pTZ57R/T and pET32a (+) vectors, respectively. Recombinant vector was transferred into E. coli BL21 CodonPlus (DE3). Protein expression was investigated by 1 mM IPTG induction. Hens were immunized by the purified recombinant FanC protein. The activity and specificity of the IgY antibody were detected by dot-blotting, Western blotting, and indirect ELISA.
  
  
  RESULTS
  We obtained FanC specific IgYs by immunizing the hens with the recombinant FanC protein. The anti-FanC IgY showed binding specifically to the FanC protein of ETEC.
  
  
  CONCLUSIONS
  The results emphasize that specific IgY against the recombinant FanC protein could be recommended as a candidate for passive immunization against ETEC infection in animals and humans.
authors.affiliation: Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.
authors.forename: Khadijeh
authors.initials: K
authors.lastname: Nasiri
authors.affiliation: Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Mashhad, Iran.
authors.forename: Saeed
authors.initials: S
authors.lastname: Zibaee
authors.affiliation: Department of Animal Science and Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
authors.forename: Mohammadreza
authors.initials: M
authors.lastname: Nassiri
authors.affiliation: Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Tahmoorespur
authors.affiliation: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Haghparast
keywords: Enterotoxigenic Escherichia coli; IgY; Immunization; Recombinant protein