Prokaryotic Expression and Initial Antigenicity Analysis of Novel Duck Reovirus σC Protein

To obtainσC protein of Novel Duck Reovirus(NDRV)from prokaryotic expression system and initial identification of its antigenicity,theσC gene of NDRV was amplified by RT-PCR from NDRV NP03 strain using the specific primers which were designed according to the published cDNA sequence.Subsequently,the target gene was cloned into pET-30a(+)prokaryotic expression vector to construct the recombinant plasmid pET-NDRV-σC,which was transformed into BL21(DE3)and the expression of theσC gene was induced(1.0mmol·L-1 IPTG,37℃).The fusion protein was identified by SDS-PAGE and purified.The antigenicity of the purified protein was characterized by Western blotting.TheσC gene was obtained with an identical sequence of 966bp to that retrieved in GenBank.The prokaryotic expression vector forσC gene was successfully constructed as confirmed by enzyme digestion and DNA sequencing.The result of SDS-PAGE showed that the fusion protein had a relative molecular weight of 41.3 ku.Moreover,theσC protein obtained form prokaryotic expression system possessed good immunoreactivity which was showed by western blotting.Together,our work laid foundation for further study of NDRVσC protein.