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PMID: 3486414 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Selection and amplification of heterologous genes encoding adenosine deaminase in mammalian cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 10 ·1986-05-00 ·Pages 3136-40

Kaufman RJ, Murtha P, Ingolia DE, Yeung CY, Kellems RE

Abstract

We demonstrate that an adenosine deaminase (ADA) cDNA gene can function as a dominant selectable and amplifiable marker for gene transfer experiments in mammalian cells. Cells that incorporate the gene can be selected by growth in the presence of low concentrations of the ADA inhibitor 2'-deoxycoformycin with cytotoxic concentrations of adenosine or its analogue 9-beta-D-xylofuranosyl adenine. The DNA copy number of the transfected ADA minigene in the isolated transformants of Chinese hamster ovary cells can be amplified greater than 100-fold by growth in ADA selection media and increasing concentrations of 2'-deoxycoformycin. This selection scheme may allow for the introduction and subsequent amplification of heterologous DNA in a variety of mammalian cells.

MeSH Terms
Adenosine/analogs & derivatives,pharmacology Adenosine Deaminase/genetics Animals Cloning, Molecular Coformycin/analogs & derivatives,pharmacology Cricetinae Drug Resistance Gene Amplification Gene Expression Regulation Genetic Vectors Mice Nucleoside Deaminases/genetics Pentostatin RNA, Messenger/genetics
Chemicals
RNA, Messenger Coformycin Pentostatin 9-xylosyladenine Nucleoside Deaminases Adenosine Deaminase Adenosine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kaufman R J
Murtha P
Ingolia D E
Yeung C Y
Kellems R E
References (41)
41 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.83.10.3136
Published
1986-05-00
Pages
3136-40
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC323467
Subset
IM
Grants
PHS HHS · A120402 · United States
NCI NIH HHS · CA00828 · United States
NCRR NIH HHS · RR05425 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.10.3136 PMC Full Text View journal details
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