Cloning, expression and immunocharacterization of the capsid protein of human Norwalk virus Guangzhou strain NVgz01.
- Author:
Xiao LI
1
;
Rong ZHOU
;
Long-bo HU
;
Xin-gui TIAN
;
Jia-yu ZHONG
;
Hui-ying SHENG
;
Chang-bing WANG
;
You-shao WANG
Author Information
- Publication Type:Journal Article
- MeSH: Blotting, Western; Capsid Proteins; analysis; biosynthesis; genetics; immunology; Cloning, Molecular; Enzyme-Linked Immunosorbent Assay; Gene Expression; Humans; Norwalk virus; genetics; Plasmids; genetics
- From: Journal of Southern Medical University 2007;27(9):1410-1413
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone, express and characterize the capsid protein of human Norwalk virus Guangzhou strain NVgz01.
METHODSOn the basis of successful construction of full-genome clones and sequence analysis of human norovirus Guangzhou strain NVgz01, the full capsid gene was ligated into pET28a (+) for expression. After IPTG induction, the recombinant protein was purified through metal (Ni(2+)) chelating affinity chromatography. Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to determine the antigenicity of the recombinant protein.
RESULTSThe recombinant capsid gene was overexpressed in E.coli, yielding the recombinant protein with relative molecular mass of 62x10(3) that was highly purified through metal (Ni(2+)) chelating affinity chromatography. IDEIA Norovirus Kit and immunoassay showed that the recombinant protein had good antigenicity.
CONCLUSIONThe capsid gene of norovirus Guangzhou strain has been cloned and expressed, which can be useful for developing diagnostic reagents or vaccine of norovirus.
