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Production of specific IgY antibody to the recombinant FanC protein produced in Escherichia coli.

Posted by on in 2016
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Nasiri K1Zibaee S2Nassiri M3Tahmoorespur M1Haghparast A42016. Iran J Basic Med Sci. 19(8):883-889.

1Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.
2Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Mashhad, Iran.
3Department of Animal Science and Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
4Department of Veterinary Medicine, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.

Abstract

OBJECTIVES:

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.

MATERIALS AND 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.

CONCLUSION:

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.

KEYWORDS:

Enterotoxigenic Escherichia coli; IgY; Immunization; Recombinant protein

PMID:
 
27746871
 
PMCID:
 
PMC5048125
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