Preparation and characteristics of polyclonal antibody against S10 encoded protein of grass carp reovirus HZ08 strain

To explore the probable function of segment 10(S10)encoded protein of grass carp reovirus(GCRV)HZ08 strain,the ORF of S10 gene was amplified and cloned into the prokaryotic expression vector pET-32a(+),and the obtained recombinant expression vector was named pET32a-S10.The recombinant vectors were transformed into BL21 competent cells.The Escherichia coli containing recombinant vector expressed fusion protein of approximately 53 ku after induction by IPTG.The recombinant protein was purified through Ni-chelating affinity chromatography,and the purity was above 97.4% explained by SDS-PAGE gel scan analysis.Polyclonal antibody against S10 encoded protein was generated by immunization of female Kunming mice with purified recombined protein.The antibody titers in sera of the immunized mice were detected by ELISA and the specificity of the antibody was identified by Western blot and indirect immunefluorescence assay(IFA).The results showed that the antibody titer cloning is 1：10⁶ and can specially identify the GCRV-HZ08.This means the S10 encoded protein is one of the structural protein of GCRV-HZ08.