Identification of the thymidine kinase gene on the genome of herpesvirus of turkeys.

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Herpesvirus of turkeys(HVT) has been used as a vaccine against Marek's disease of chickens. The live vaccine could be a candidate for constructing of live recombinant vaccine. The purpose for this research is to determine the localization of the thymidine kinase(TK) gene on the HVT genome. I have constructed the restriction enzyme fragment maps of HVTDNA in collaboration with Dr Nonoyama group at Showa University Research Institute for Medicine in Florida. Then, we attempted to localize the TK gene of HVT on the viral genome by Southern blot hybridization using herpes simple virus type 1(HSV1) TK gene as a probe. The HSV1 TK gene hybridized in the low stringent conditions to Pst-A and Hind -E fragments of HVTDNA. These fragments overlap within the long unique sequence of HVTDNA. Next, we also attempted to detect the virus-specific TK activity in HVT infected TK^- cells. The HVT-induced TK activity was observed in virus infected cells and the in vitro activity was significantly inhibited in the presence of 0.2mMTTP. In addition, the HVTDNA replication was inhibited by acyclovir, which was known to be phosphorylated by herpesvirus-specific TK. These results suggests that the TK activity induced by HVT is virus specific. We need to determine the nucleotide sequences of the HVT-TK gene and to characterize the protein encoded from the gene.