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PMID: 6296831 Published · ppublish English Journal Article

Vaccinia virus: a selectable eukaryotic cloning and expression vector.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 23 ·1982-12-00 ·Pages 7415-9

Mackett M, Smith GL, Moss B

Abstract

Foreign DNA was inserted into two nonessential regions of the vaccinia virus genome by homologous recombination in cells infected with virus and transfected with plasmids containing the foreign DNA elements flanked by vaccinia virus DNA. Thymidine kinase-negative (TK-) recombinants were selected after inserting foreign DNA into the coding region of the TK gene of wild-type vaccinia virus; TK+ recombinants were selected after inserting the herpesvirus TK gene into TK- mutants of vaccinia virus. For TK+ expression, it was necessary to insert a 275-base-pair DNA fragment containing the initiation site and sequences upstream of an early vaccinia virus transcript next to the coding sequences of the herpesvirus gene. The unique ability of the herpesvirus TK to phosphorylate 125I-labeled deoxycytidine provided independent confirmation of gene expression. These studies demonstrate the use of vaccinia virus as a selectable cloning and expression vector, confirm the map location of the vaccinia virus TK gene, and provide initial information regarding the location of vaccinia virus transcriptional regulatory sequences.

MeSH Terms
Cloning, Molecular/methods Gene Expression Regulation Genes Genetic Vectors Herpesviridae/genetics Thymidine Kinase/genetics Transcription, Genetic Transfection Vaccinia virus/genetics
Chemicals
Thymidine Kinase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mackett M
Smith G L
Moss B
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36 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.79.23.7415
Published
1982-12-00
Pages
7415-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC347350
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.79.23.7415 PMC Full Text View journal details
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