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PMID: 6933469 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 77 ·No. 7 ·1980-07-00 ·Pages 4216-20

Urlaub G, Chasin LA

Abstract

Mutants of Chinese hamster ovary cells lacking dihydrofolate reductase (tetrahydrofolate dehydrogenase, 7,8-dihydrofolate:NADP+ oxidoreductase; EC 1.5.1.3) activity were isolated after mutagenesis and exposure to high-specific-activity [3H]deoxyuridine as a selective agent. Fully deficient mutants could not be isolated starting with wild-type cells, but could readily be selected from a putative heterozygote that contains half of the wild-type level of dihydrofolate reductase activity. The heterozygote itself was selected from wild-type cells by using [3H]deoxyuridine together with methotrexate to reduce intracellular dihydrofolate reductase activity. Fully deficient mutants require glycine, a purine, and thymidine for growth; this phenotype is recessive to wild type in cell hybrids. Revertants have been isolated, one of which produces a heat-labile dihydrofolate reductase activity. These mutants may be useful for metabolic studies relating to cancer chemotherapy and for fine-structure genetic mapping of mutations by using available molecular probes for this gene.

MeSH Terms
Animals Cell Line Cricetinae Cricetulus Female Heterozygote Hybrid Cells/enzymology Mutation Ovary Selection, Genetic Tetrahydrofolate Dehydrogenase/deficiency,genetics
Chemicals
Tetrahydrofolate Dehydrogenase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Urlaub G
Chasin L A
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.77.7.4216
Published
1980-07-00
Pages
4216-20
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC349802
Subset
IM
Grants
NIGMS NIH HHS · GM-22629 · United States
External Links
PubMed source DOI: 10.1073/pnas.77.7.4216 PMC Full Text View journal details
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