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Phylogenetic Analysis of the DNA Polymerase Gene of a Novel Alphaherpesvirus Isolated from an Indian <Emphasis Type="Italic">Gyps</Emphasis> Vulture
Authors:Maria?Cardoso  Alex?Hyatt  Paul?Selleck  Sue?Lowther  Vibhu?Prakash  Deborah?Pain  Andrew?A?Cunningham  Email author" target="_blank">David?BoyleEmail author
Institution:(1) Australian Animal Health Laboratories, CSIRO Livestock Industries, Private Bag 24, Geelong, Victoria, 3220, Australia;(2) Bombay Natural History Society, Hornbill House, S. B. Singh Road, Mumbai, 400 023, India;(3) RSPB, The Lodge, Sandy, Bedfordshire, SGI9 2DL, UK;(4) Institute of Zoology, Zoological Society of London, Regentrsquos Park, London, NW1 4RY, United Kingdom
Abstract:The DNA polymerase gene of a novel herpesvirus, vulture herpesvirus (VHV), isolated from an Indian Gyps vulture was completely sequenced using primer walking and transposon insertion strategies. DNA sequencing analysis revealed a single open reading frame (ORF) of 3660 nucleotides (53% G-C content) able to encode 1219 amino acids. Identification was based on a nucleotide sequence identity of approximately 50% to other herpesvirus sequences found in Genbank. Nine motifs were identified that are conserved amongst all known herpesviruses and are found within the 3prime–5prime exonuclease and DNA binding functional domains of the DNA polymerase enzyme. Phylogenetic analysis using Clustal W with neighbour-joining revealed VHV to group within the subfamily Alphaherpesvirinae, more closely related to the avian herpesviruses than to those of other species. Partial sequence data also revealed VHV to contain other genes fundamental to the structure and replication of all herpesvirus genomes. A Real Time PCR Taqman assay specific for the VHV DNA polymerase gene was designed to detect the presence of VHV genomic material in post mortem tissue samples from diseased birds. Positive tissues included the spleen, rectum, thymus, kidney and brain. A herpesvirus specific to vultures may pose a threat to the management of captive breeding programs being established to assist the survival of wild populations of Gyps vultures.
Keywords:alphaherpesvirus  DNA polymerase  Gyps vultures  Real Time PCR  phylogeny  sequencing
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