Cloning, expression and antigenic analysis of VP1-VP4 gene encoding the structural protein of Coxsackie virus A16.
- Author:
Yuanbin SONG
1
;
Sijie HE
;
Nan YU
;
Xinxin CHEN
;
Bin WANG
;
Xiaoyan CHE
;
Qiyi ZENG
Author Information
- Publication Type:Journal Article
- MeSH: Antigens, Viral; genetics; immunology; Capsid Proteins; classification; genetics; immunology; Cloning, Molecular; Enterovirus A, Human; genetics; Gene Expression; Genetic Vectors; Plasmids; Recombinant Proteins; genetics; immunology; Reverse Transcriptase Polymerase Chain Reaction
- From: Journal of Southern Medical University 2012;32(12):1713-1717
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone and express VP1-VP4 genes encoding the structural proteins of Coxsackie virus A16 and analyze the antigenicity of the expressed recombinant proteins.
METHODSThe VP1-VP4 cDNAs were amplified with RT-PCR from the extracted viral RNA and cloned into pMD19-T vectors. The VP1-VP4 genes were inserted to the multi-cloning sites of the plasmid pQE30a, and the protein expressions in E. coli M15 were induced by IPTG. After purification by washing with 8 mol/L urea under denaturing condition, the recombinant proteins were identified by Western blotting and ELISA for their immunogenicity against rabbit antisera of Coxsackie virus A16 and enterovirus 71, respectively.
RESULTSThe recombinant VP1-VP4 proteins were highly expressed in E. coli M15. The purified proteins could be recognized by rabbit antiserum of Coxsackie virus A16 and showed cross reactivity with the rabbit antiserum of Enterovirus 71.
CONCLUSIONThe recombinant Coxsackie virus A16 VP1-VP4 proteins obtained possess good antigenicity.
