Sequence determination of variable regions within the genomes of gallid herpesvirus-2 pathotypes

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Abstract
Comparative genomic studies of attenuated and virulent strains of Gallid herpesvirus 2 (GaHV-2) have identified 6 regions of sequence variability. These regions include the open reading frames (ORFs) encoding UL36 and UL49 and regions devoid of large ORFs (132-bp repeats, a-like sequences and the junctions flanking the unique short region). Our data indicate that the carboxyl terminus of UL36 contains regions of heterogeneity that are unique to CVI988-derived attenuated strains. A deletion of the TKSERT domain and a glycine245 polymorphism in the UL49 proteins were also identified in these derivatives. Phylogenetic analyses of both UL36 and UL49 sequences indicate that CVI988-derived strains partition differently from other attenuated strains (RM-1 and R2/23), indicating that additional mutations contribute to attenuation. In very virulent and very virulent plus strains a single nucleotide polymorphism (SNP) was identified within the 132-bp tandem repeats. Within the junctions flanking the unique short region, these strains also contain deletions in sequences that are predicted to bind the transcription factor NF kappaB. In some attenuated strains, deletions were also identified in the latency-associated transcript (LAT) promoters and adjacent regions encoding microRNAs. These results indicate that virulence is likely multi-factorial with contributions from both multiple genes and cis-acting sites.

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