Construction and characterization of a bovine herpesvirus 5 mutant with a deletion of the gI, gE and US9 genes

Ana Cláudia Franco, Sílvia De Oliveira Hübner, Anna Paula De Oliveira, Helena B.De C.Ruthner Batista, Paulo Michel Roehe, Franciscus A.M. Rijsewijk

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


Bovine herpesvirus 5 (BoHV-5) is a important cause of viral encephalitis in cattle in South America. Within the framework of developing a differential vaccine against BoHV-5, a deletion mutant was constructed based on a Brazilian BoHV-5 isolate. The target of the deletions were genes that code proteins implicated in the neurovirulence of BoHV-5, the glycoprotein I (gI), glycoprotein E (gE) and membrane protein US9. To construct the deletion mutant of BoHV-5, the flanking regions of all three genes were cloned in a prokaryotic plasmid. This deletion fragment was co-transfected with the viral DNA into bovine cells. Identification of deletion mutants was performed by immunostaining with an anti-gE monoclonal antibody. One of the gE negative viral populations found was purified, amplified and further examined by restriction endonuclesase analysis of its genomic DNA. The plaque sizes and penetration kinetics of the deletion mutant and wild type viruses were compared. The plaque sizes of the deletion mutant were significantly smaller than those of the parental strain (p ≤ 0.05), but no statistical differences were observed in penetration kinetics. The results indicate that the gI/ gE/US9 deletion mutant of BoHV-5 may have a reduced virulence in the host and is still viable enough to be a good candidate for the development of a BoHV-5 vaccine.

Original languageEnglish (US)
Pages (from-to)667-673
Number of pages7
JournalBrazilian Journal of Microbiology
Issue number4
StatePublished - 2007
Externally publishedYes


  • BoHV-5
  • Bovine herpesvirus 5
  • Deletion mutant
  • Glycoprotein genes

ASJC Scopus subject areas

  • Microbiology


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