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The essential and non-essential genes of Bovine herpesvirus 1

Fiona M. McCarthy^1,

¹ Department of Primary Industries and Fisheries, St Lucia, Brisbane, QLD, Australia
² School of Veterinary Science, University of Queensland, St Lucia, Brisbane, QLD, Australia

Correspondence
Timothy J. Mahony
Timothy.Mahony{at}dpi.qld.gov.au

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.

Present address: Department of Basic Sciences and Institute for Digital Biology, College of Veterinary Medicine, Mississippi State University, PO Box 1600, MS 39762, USA.

Two supplementary tables detailing primer sequences are available with the online version of this paper.