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PMID: 6283542 Published · ppublish English Journal Article

DNA-mediated gene transfer: recombination between cotransferred DNA sequences and recovery of recombinants in a plasmid.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 9 ·1982-05-00 ·Pages 2748-52

Anderson RA, Krakauer T, Camerini-Otero RD

Abstract

Ltk- aprt- mouse L cells were transformed to the tk+ phenotype with 10 ng of the herpes simplex virus-1 thymidine kinase (tk) gene and 20 micrograms of pBR322 or simian virus 40 (SV40) DNA. DNAs from five cloned cell lines show restriction endonuclease fragments that hybridize to both tk and pBR322 or SV40 DNA. In all of the cell lines some of these fragments also contain cellular DNA sequences. The use of carrier DNAs with defined sequences has enabled us to demonstrate that the joining of carrier and selectable gene sequences occurs in mouse cells. In one case we have been able to use the ampicillin resistance marker of pBR322 to "rescue" a recombinant plasmid. An analysis of the junction between pBR322 and tk in this plasmid suggests that a small area of homology (16 of 19 base pairs) might be involved in the recombination process.

MeSH Terms
Animals Cells, Cultured Genes, Viral Mice Plasmids Recombination, Genetic Simplexvirus/enzymology Thymidine Kinase/genetics Transformation, Genetic
Chemicals
Thymidine Kinase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Anderson R A
Krakauer T
Camerini-Otero R D
References (34)
34 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.79.9.2748
Published
1982-05-00
Pages
2748-52
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC346283
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.79.9.2748 PMC Full Text View journal details
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