The essential and non-essential genes of Bovine herpesvirus 1

J Gen Virol. 2008 Nov;89(Pt 11):2851-2863. doi: 10.1099/vir.0.2008/002501-0.

Abstract

Bovine herpesvirus 1 (BoHV-1) is an economically important pathogen of cattle associated with respiratory and reproductive disease. To further develop BoHV-1 as a vaccine vector, a study was conducted to identify the essential and non-essential genes required for in vitro viability. Random-insertion mutagenesis utilizing a Tn5 transposition system and targeted gene deletion were employed to construct gene disruption and gene deletion libraries, respectively, of an infectious clone of BoHV-1. Transposon insertion position and confirmation of gene deletion were determined by direct sequencing. The essential or non-essential requirement of either transposed or deleted open reading frames (ORFs) was assessed by transfection of respective BoHV-1 DNA into host cells. Of the 73 recognized ORFs encoded by the BoHV-1 genome, 33 were determined to be essential and 36 to be non-essential for virus viability in cell culture; determining the requirement of the two dual copy ORFs was inconclusive. The majority of ORFs were shown to conform to the in vitro requirements of BoHV-1 homologues encoded by human herpesvirus 1 (HHV-1). However, ORFs encoding glycoprotein K (UL53), regulatory, membrane, tegument and capsid proteins (UL54, UL49.5, UL49, UL35, UL20, UL16 and UL7) were shown to differ in requirement when compared to HHV-1-encoded homologues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • DNA, Viral / genetics
  • DNA, Viral / isolation & purification
  • Gene Deletion
  • Gene Expression Profiling
  • Gene Library
  • Genes, Essential*
  • Genes, Viral*
  • Genome, Viral
  • Herpesvirus 1, Bovine / genetics*
  • Herpesvirus 1, Bovine / immunology
  • Herpesvirus 1, Human / genetics
  • Humans
  • Open Reading Frames
  • Polymerase Chain Reaction
  • Restriction Mapping
  • Transfection
  • Viral Vaccines
  • Virus Replication / genetics

Substances

  • DNA, Viral
  • Viral Vaccines