Because newly identified green turtle herpesvirus (GTHV) is associated pathogenically with marine turtle fibropapillomatosis (FP) and it has not been isolated in vitro, molecular sequencing and analysis of the genomic DNA of this putative reptilian herpesvirus will enhance the current understanding of GTHV in causing the FP disease. An inverse polymerase chain reaction (IPCR) genomic walking technique was developed to obtain new DNA sequences based on a portion of known genomic sequence. Through two genomic walks, a 2169 bp DNA fragment of GTHV was cloned and sequenced. Sequence analysis shows that this DNA fragment contains the entire gene of the UL28, as well as the partial genomic sequence of the UL27 gene. The UL28 gene is 2250 bp long and encodes a 750-amino acid peptide known as ICP18.5 assembly protein of herpesviruses. Phylogenetic analysis of the GHTV UL28 gene showed a high sequence homology with the UL28 homologs of other herpesviruses and supports the current classification of GTHV to be a member of Alphaherpesvirinae. Identification of the,genomic sequences of GTHV provides a molecular base for the development of diagnostic immunoassay and also for the determination of the pathogenic role of GTHV infection. (C) 2004 Published by Elsevier B.V.
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