J Korean Soc Virol.
1997 Dec;27(2):105-113.
Overexpression of the E1193-283 find E2384-649 Proteins of Hepatitis C Virus in GST Fusion Forms in E. coli and Their Immunogenicity
Abstract
- The truncated E1192-283 and E2384-649 genes of hepatitis C virus (HCV) linked to the gene for glutathione 5-transferase (GST) were constructed and their expressions were analyzed. The GST-E1192-283 fusion gene overexpressed the fusion protein in E. coli as a soluble form, while the GST-E1192-383 plasmid did not express expected fusion protein. The purified GST-E1192-283 fusion protein was efficiently cleaved by thrombin. More than 90% pure, HCV E1192-283 protein was obtained by GST-agarose chromatography. The truncated GST-E2384-649 fusion gene expressed the fusion protein mainly as an insoluble form, whereas the GST-E2384-740 did not express the fusion protein. The truncated GST-E1 182-283 and GST-E2384-649 fusion proteins reacted specifically with an HCV patient serum. In addition, mice immunized with either the purified E1192-283 or GST-E2384-649 proteins generated specific antibodies to each antigen. The results suggested that hydrophobic carboxyl portions of the E1 and E2 proteins might affect expression levels as well as the solubility of each fusion protein in bacteria. Also, the truncated E1 protein with Tyr-192 to Ser-283 contained antigenic epitope(s) which could be specifically recognized by an HCV patient serum.
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