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PMID: 6289324 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Construction of poxviruses as cloning vectors: insertion of the thymidine kinase gene from herpes simplex virus into the DNA of infectious vaccinia virus.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 16 ·1982-08-00 ·Pages 4927-31

Panicali D, Paoletti E

Abstract

We have constructed recombinant vaccinia viruses containing the thymidine kinase gene from herpes simplex virus. The gene was inserted into the genome of a variant of vaccinia virus that had undergone spontaneous deletion as well as into the 120-megadalton genome of the large prototypic vaccinia variant. This was accomplished via in vivo recombination by cotransfection of eukaryotic tissue culture cells with cloned BamHI-digested thymidine kinase gene from herpes simplex virus containing flanking vaccinia virus DNA sequences and infectious rescuing vaccinia virus. Pure populations of the recombinant viruses were obtained by replica filter techniques or by growth of the recombinant virus in biochemically selective medium. The herpes simplex virus thymidine kinase gene, as an insert in vaccinia virus, is transcribed in vivo and in vitro, and the fidelity of in vivo transcription into a functional gene product was detected by the phosphorylation of 5-[125I]iodo-2'-deoxycytidine.

MeSH Terms
Base Sequence Cloning, Molecular/methods Genetic Vectors Simplexvirus/genetics Thymidine Kinase/genetics Vaccinia virus/genetics
Chemicals
Thymidine Kinase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Panicali D
Paoletti E
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40 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.79.16.4927
Published
1982-08-00
Pages
4927-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC346798
Subset
IM
Grants
NIGMS NIH HHS · GM23853 · United States
External Links
PubMed source DOI: 10.1073/pnas.79.16.4927 PMC Full Text View journal details
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