Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpesviruses.
Plaques formed by herpes simplex virus (HSV), pseudorabies virus, and varicella-zoster virus were studied by plaque autoradiography after [14C]thymidine labeling. Standard thymidine kinase-positive (TK+) viruses and TK- mutants of HSV types 1 and 2 and pseudorabies virus were studied, including cell cultured viruses and viruses isolated from animals. Autoradiography was performed with X-ray film with an exposure time of 5 days. After development of films, TK+ plaques showed dark rims due to isotope incorporation, whereas TK- plaques were minimally labeled. Plaque autoradiography of stock TK- viruses showed reversion frequencies to the TK+ phenotype of less than 10(-3). Autoradiography indicated that TK- virus retained the TK- phenotype after replication in vivo. In addition, it was shown that TK- HSV could be isolated from mouse trigeminal ganglion tissue after corneal inoculation of TK- HSV together with TK+ HSV. The plaque autoradiographic procedure was very useful to evaluate proportions of TK+ and TK- virus present in TK+-TK- virus mixtures.
D000818:Animals; D001345:Autoradiography; D005260:Female; D006564:Herpesviridae / Q000201:enzymology* / Q000472:pathogenicity; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D013936:Thymidine / Q000378:metabolism; D013937:Thymidine Kinase / Q000032:analysis*; D014316:Tritium; D010948:Viral Plaque Assay
D014316:Tritium; D013937:Thymidine Kinase; D013936:Thymidine