[Cloning, expression and immunocharacterization of the capsid protein of human Norwalk virus Guangzhou strain NVgz01].
OBJECTIVE
To clone, express and characterize the capsid protein of human Norwalk virus Guangzhou strain NVgz01.


METHODS
On 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.


RESULTS
The 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.


CONCLUSIONS
The capsid gene of norovirus Guangzhou strain has been cloned and expressed, which can be useful for developing diagnostic reagents or vaccine of norovirus.
Li
Xiao
X
South China Institute of Oceanology, Guangzhou 510301, China. xiao-2001@21cn.com
Zhou
Rong
R
Hu
Long-bo
LB
Tian
Xin-gui
XG
Zhong
Jia-yu
JY
Sheng
Hui-ying
HY
Wang
Chang-bing
CB
Wang
You-shao
YS
D015153:Blotting, Western; D036022:Capsid Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D006801:Humans; D009663:Norwalk virus / Q000235:genetics*; D010957:Plasmids / Q000235:genetics
D036022:Capsid Proteins