Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Sequencing and Baculovirus-Based Expression of the Glycoprotein B2 Gene of HSV-2 (G)

  • Uh, Hong-Sun (Department of Biological Sciences, Konkuk University) ;
  • Park, Jong-Kuk (Department of Biological Sciences, Konkuk University) ;
  • Kang, Hyun (Department of Biological Sciences, Konkuk University) ;
  • Kim, Soo-Young (Department of Biological Sciences, Konkuk University) ;
  • Lee, Hyung-Hoan (Department of Biological Sciences, Konkuk University)
  • Published : 2001.06.01
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Abstract

The gene for glycoprotein B (gB2) of HSV-2-strain G was subcloned, sequenced, recombinated into the lacZ-HcNPV, expressed in insect cells, and compared with the homologous gene of other HSV-2 strains. The ORF of the gB2 gene was 2,715 bp. The overall nucleotide sequence homology of te gB2 gene compared ith that of the two previously reported HSV-2 strains appeared to be over 98%. A recombinant virus named Baculo-gB2 protein in insect cells. The recombination was confirmed by a PCR and the expression was demonstrated by radio immunoprecipitation. Insect cells infected with the Baculo-gB2 virus synthesized and processed gB2 with approximately 120 kDa in the cells, and then secreted it into the culture media, where it reacted with a nomoclonal antibody to gB2. The gB2 polypeptide contained two main hydrophobic regions (a signal sequence from 1 to 23 amino acid residues, and a membrane anchor sequence from aa 745 to 798), eight N-glycosylation sites evenly distributed, and was rich in alanine (11.2%). Antibodies to this recombinant protein that were raised in mice recognized the viral gB2 and neutralized the infectivity of the HSV-2 in vitro. There results show that the gB2 protein was successfully porduced in insect cells and could be used to raise a protective neutralizing antibody. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

Keywords

References

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