The expression and purification of VP8 * proteins of the rotavirus vaccine strains
10.3760/cma.j.issn.1003-9279.2015.06.019
- VernacularTitle:轮状病毒疫苗株VP8*核心区蛋白的表达和纯化
- Author:
Xiaoman SUN
1
;
Nijun GUO
;
Dandi LI
;
Zhaojun DUAN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Rotavirus;
Vaccine strain;
VP8 * core protein;
Protein expression
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(6):543-546
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the functional and structural character of VP8 * proteins of the RV vaccine strains,VP8 * core proteins were expressed and purified.Methods The LLR RNA was extracted directly from the vaccine oral liquid,followed by RT-PCR.The VP8 * core fragment was obtained by PCR with specific primers.The full length genes of Rotateq and Rotarix VP8 * were synthesized and the VP8 * core fragments were obtained by PCR.The VP8 * core fragments were cloned to the pET30a vector respectively.The recombinant plasmids were transformed to the BL21 competent cells to do the protein expression.Then the proteins were further purified by affinity chromatography and gel filtration.The protein samples were evaluated by SDS-PAGE.Results The VP8 * core proteins were expressed in soluble form and purified and the protein bands showed at about 20 kDa in the SDS-PAGE.Conclusions The VP8 * core-pET30a plasmids were constructed and the VP8 * core proteins of the RV vaccine strains were successfully expressed and purified,which may provide the basis for the characterization of receptor binding specificity of the RV vaccine strain and the evaluation of the vaccine efficacy.
