Application and expression of HSV gG1 protein from a recombinant strain.

Abstract

According to the homologous sequence of glycoprotein G1 (gG1) genes from different strains of herpes simplex virus type 1 (HSV-1), a pair of primers was designed to amplify the gG1 gene fragment by PCR. Both the PCR product and the pGEX-4T-1 vector were digested with EcoR I and Sal I. The gG1 gene fragment was subcloned into the digested pGEX-4T-1 vector to construct a recombinant plasmid (pGEX-4T-1-gG1). The resultant plasmid was identified by dual-enzyme digestion and sequence analysis, and then transformed into Escherichia coli BL21 for expression under the induction of isopropyl β-D-1-thiogalactoside (IPTG). The expressed GST-gG1 fragment was detected by SDS-PAGE and purified by affinity chromatography. The properties of GST-gG1 fragment were evaluated by immunoblot analysis. Enzyme-linked immunosorbent assays (ELISAs) based on the GST-gG1 fragment were used for determining IgG or IgM to HSV-1. The GST-gG1 fragment-specific ELISA was also compared with ELISA with whole-HSV-1 antigen and commercial ELISA kits. The gG1-specific IgG and IFN-γ producing CD8+ T cells were induced in mice immunized with the GST-gG1 fragment. These results indicated that the GST-gG1 fragment could be used for replacing whole-virus antigen to detect IgM and IgG to HSV-1 in human sera, which provided a strategy for developing vaccines to protect HSV-1 infection using gG1 fragment.

• Authors: Hua Yan, Huishen Yan, Tao Huang, Guocai Li, Weijuan Gong, Hongmei Jiao, Hongju Chen, Mingchun Ji
• Journal: Journal of virological methods (J Virol Methods) Vol. 169 Issue 2 Pg. 351-8 (Nov 2010) ISSN: 1879-0984 [Electronic] Netherlands
• PMID: 20705103 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2010 Elsevier B.V. All rights reserved.

Chemical References

• Antibodies, Viral
• DNA, Viral
• Immunoglobulin G
• Immunoglobulin M
• Recombinant Proteins
• Viral Envelope Proteins
• glycoprotein gG-1, herpes simplex virus type 1

Topics

• Animals
• Antibodies, Viral (blood)
• Chromatography, Affinity
• Cloning, Molecular
• DNA, Viral (genetics)
• Enzyme-Linked Immunosorbent Assay (methods)
• Escherichia coli (genetics)
• Gene Expression
• Herpesvirus 1, Human (genetics)
• Humans
• Immunoglobulin G (blood)
• Immunoglobulin M (blood)
• Mice
• Mice, Inbred BALB C
• Plasmids
• Recombinant Proteins (biosynthesis, genetics, isolation & purification)
• Viral Envelope Proteins (biosynthesis, genetics, isolation & purification)
• Virology (methods)