[Expression and immunological reactivity of recombinant HCV-core protein].
OBJECTIVE
To express HCV-core proteins in E.coli and to develop effective HCV-core DNA-based vaccine.


METHODS
The vector that expresses the highly conserved HCV core genes were constructed. The pGEX-3X HCVCore constructs contained the 1-201 ncls (1-67aa, C201), 1-402 ncls (1-134aa, C402) and 1-591ncls ( 1-197aa, C591), then expressed in E.coli cells.


RESULTS
The products of HCV C201 and C402 genes were expressed as a fusion protein with glutathione-S-transferase (GST, 26kDa) whose molecular weight were 3.1 x 10(4) and 3.9 x 10(4) separately. C591 gene was not effectively expressed in E.coli. The expressed proteins were sequestered within inclusion bodies (IB) and a variety of procedures designed to minimize IB formation proved unsuccessful. The method finally adopted involved the purification of inclusion bodies followed by the solubilization, purification, and refolding of the expressed protein. The purified C402 protein was antigenically reactive with serum from chronically infected HCV patients. BALB/C mice were immunized by a subcutaneous injection of C402 protein together with Freund's complete adjuvant which produced strong anti-HCV core humoral immune responses.


CONCLUSIONS
It is important for the study of gene vaccine to construct a certain length of HCV core gene.
Dai
W
W
Shenzhen East Lake Hospital, Shenzhen 518020, China.
Ma
W
W
Guo
Y
Y
D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D014758:Viral Core Proteins / Q000096:biosynthesis* / Q000276:immunology
D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus