Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpes viruses

Richard Tenser, J. C. Jones, S. J. Ressel, F. A. Fralish

Research output: Contribution to journalArticle

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Abstract

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 <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.

Original languageEnglish (US)
Pages (from-to)122-127
Number of pages6
JournalJournal of clinical microbiology
Volume17
Issue number1
StatePublished - Jan 1 1983

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Thymidine Kinase
Autoradiography
Thymidine
Viruses
Simplexvirus
Suid Herpesvirus 1
Phenotype
Trigeminal Ganglion
X-Ray Film
Human Herpesvirus 3
Human Herpesvirus 2
Human Herpesvirus 1
Isotopes
Cultured Cells

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

Cite this

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title = "Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpes viruses",
abstract = "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 <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.",
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Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpes viruses. / Tenser, Richard; Jones, J. C.; Ressel, S. J.; Fralish, F. A.

In: Journal of clinical microbiology, Vol. 17, No. 1, 01.01.1983, p. 122-127.

Research output: Contribution to journalArticle

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T1 - Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpes viruses

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AU - Ressel, S. J.

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N2 - 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 <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.

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