Analysis of soluble expression,purification and antigenic characterization of P protein of GⅡ.2[P16]norovirus
10.13200/j.cnki.cjb.003935
- VernacularTitle:GⅡ.2[P16]型诺如病毒P蛋白的可溶表达、纯化及其抗原特性分析
- Author:
LI Pan
- Publication Type:Journal Article
- Keywords:
Norovirus(NV);
GⅡ.2[P16];
P protein;
Soluble expression
- From:
Chinese Journal of Biologicals
2024;37(8):952-956
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo obtain the soluble expression of P protein of norovirus(NV)G Ⅱ. 2[P16]and analyze its antigenic characteristics.MethodsThe recombinant plasmid GⅡ.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 antigenantibody binding ability was identified by ELISA.ResultsThe target protein of recombinant BL21 Star(DE3)pLySs was highly expressed in soluble form. After purification by affinity chromatography,the purity of GⅡ.2 P protein was 95. 53%,and the antigenicity was good.ConclusionSoluble 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.
