Archives of Virology

, Volume 141, Issue 1, pp 57–71 | Cite as

Construction of bovine herpesvirus-1 (BHV-1) recombinants which express pseudorabies virus (PRV) glycoproteins gB, gC, gD, and gE

  • H. Otsuka
  • X. Xuan
Original Papers

Summary

We have improved the method for constructing recombinants of bovine herpesvirus type-1 (BHV-1). Using this method, we constructed three recombinants in which the pseudorabies virus (PRV) thymidine kinase (tk) gene was inserted at three different sites in the unique short region of BHV-1. These three sites are located in the open reading frame of gE, gG and gI genes. Previously, two sites (tk and gC) had been used to insert foreign DNA fragments to BHV-1 genome. Therefore we now have 5 sites in BHV-1 where DNA can be inserted. The gB, gC, gD, gE and gI genes of PRV were successfully inserted at the tk or the gC gene of BHV-1 genome and Western blot analyses confirmed that the recombinants express PRV gB, gC, gD and gE. Anti-PRV gB and gC antibodies as well as anti-PRV polyclonal serum neutralized BHV-1 recombinants which express PRV gB and gC. The latter was neutralized more strongly. However, anti-gD monoclonal antibody and anti-PRV polyclonal serum failed to neutralize gD-expressing recombinants. This suggests that PRV gC and some gB are integrated into the viral envelope of the recombinants, but very little gD is present in the viral envelope.

Keywords

Western Blot Blot Analysis Western Blot Analysis Thymidine Thymidine Kinase 

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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • H. Otsuka
    • 1
  • X. Xuan
    • 1
  1. 1.Department of Animal Resource Science, Graduate School of Agricultural SciencesUniversity of TokyoTokyoJapan

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