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KMID : 0545120000100030355
Journal of Microbiology and Biotechnology
2000 Volume.10 No. 3 p.355 ~ p.362
Expression of the HSV-1(F)Glycoprotein B Gene in Insect Cells Infected by HcNPV Recombinant
Cha, Soung Chul
Kang, Hyun/Lee, Sook Yeon/Park, Gap Ju/Lee, Hyung Hoan
Abstract
Abstract The Herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) gene in the pHLA-21 plasmid was inserted into a baculovirus (Hyphantria cunea nuclear polyhedrosis virus) expression vector (lacZ-HcNPV) to construct a recombinant virus gB-HcNPV expressing gB. Spodoptera frugiperda cells infected with this recombinant virus synthesized and processed gB of approximately 120kDa, which cross-reacted with the monoclonal antibody to gB. The recombinant gB was identified on the membrane of the insect cells using an immunofluorescence assay. Antibodies to this recombinant raised in mice recognized the viral gB and neutralized the infectivity of the HSV-1 in vitro. These results show that the gB gene has the potential to be expressed in insect cells. They also demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the lacZ-HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.
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