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PMID: 225554 Published · ppublish English Journal Article

Selective assay for herpes simplex viruses expressing thymidine kinase.

Journal of virology ·Vol. 31 ·No. 2 ·1979-08-00 ·Pages 281-7

Campione-Piccardo J, Rawls WE, Bacchetti S

Abstract

A technique for selecting herpes simplex viruses expressing the viral thymidine kinase (TK+) from a population of predominantly TK- viruses was developed. This was accomplished by infecting TK- cells and incubating the cultures under a liquid overlay medium containing methotrexate. Since the TK- cells survive in this medium for only a limited period of time, it was necessary to add fresh uninfected TK- cells 48 h after infection. The technique allowed the detection and quantitation of the TK+ virus fraction in mixtures of TK+ and TK- viruses where the TK+ fraction was present in frequencies as low as 10(-5). It was also used to estimate reversion frequencies and to obtain and analyze TK+ revertants from TK- mutant strains of herpes simplex virus type 1.

MeSH Terms
Cell Line Culture Media Humans Methotrexate Microbiological Techniques Mutation Simplexvirus/enzymology,genetics,isolation & purification Thymidine Kinase/genetics Virus Replication
Chemicals
Culture Media Thymidine Kinase Methotrexate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Campione-Piccardo J
Rawls W E
Bacchetti S
References (22)
22 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.31.2.281-287.1979
Published
1979-08-00
Pages
281-7
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC353450
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.31.2.281-287.1979 PMC Full Text View journal details
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