Preparation of Polyclonal Antibodies Against Recombinant Nucleocapsid Protein of Infectious Bronchitis Virus
The objective of this study was to prepare the murine-derived polyclonal antibodies(pAbs)against infectious bronchitis virus(IBV)N protein.The N gene was amplified by RT-PCR using IBV Beaudette genome as template and the amplified fragment was used for construction of the recombinant prokaryotic expression plasmid pET-32a-His-IBV-N.This plasmid was transformed into E.coli BL21(DE3)to express His-tagged recombinant protein N.BALB/c mice were immunized three times with the purified His-N protein emulsified in the same volume of Freund's complete adjuvant and the serum samples were collected for the following study.The specificity of the anti-N pAbs was examined in indirect immunofluorescence assay(IFA)and Western blot.The results showed that the murine anti-N pAbs specifically recognized both plasmid expressed N protein and virus expressed N protein.The availability of anti-N pAbs provided the tool for study of the biological function of N protein and offered the reagent for IB diagnosis.
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