Objective To obtain the soluble expression of P protein of norovirus(NV)G Ⅱ.2[P16]and analyze its antigenic characteristics.Methods The recombinant plasmid G Ⅱ.2-NV-pET-28a was constructed by optimizing the NV GⅡ.2[P16]P protein gene,synthesizing and cloning it into pET-28a(+)vector.The recombinant plasmid was transformed into DH5α competent cells,and the expression strain was optimized to make the P protein express in soluble form.The expressed P protein was purified by Ni-NTA affinity chromatography,of which the antigenicity was detected by Western blot,the purity was detected by size exclusion high performance liquid chromatography(SEC-HPLC),and the antigen-antibody binding ability was identified by ELISA.Results The target protein of recombinant BL21 Star(DE3)pLySs was highly expressed in soluble form.After purification by affinity chromatography,the purity of G Ⅱ.2 P protein was 95.53％,and the antigenicity was good.Conclusion Soluble NV P protein was successfully prepared by prokaryotic expression strain,and the antigenicity of P protein was confirmed,which laid a foundation of the development of NV G Ⅱ.2[P16]detection kits and the recombinant subunit vaccines.
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