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PMID: 6251468 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Transformation of mammalian cells with an amplifiable dominant-acting gene.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 77 ·No. 6 ·1980-06-00 ·Pages 3567-70

Wigler M, Perucho M, Kurtz D, Dana S, Pellicer A, Axel R, Silverstein S

Abstract

We have transferred a mutant hamster gene coding for an altered dihydrofolate reductase to wild-type cultured mouse cells by using total genomic DNA from methotrexate-resistant Chinese hamster ovary A29 cells as donor. By demonstrating the presence of hamster gene sequences in transformants we have provided direct evidence for gene transfer. Transformants selected for increased resistance to methotrexate contain increased amounts of the newly transferred gene. We have used this mutant dhfr gene to introduce the Escherichia coli antibiotic resistance plasmid pBR322 into animal cells. Amplification of the dhfr sequences results in amplification of the pBR322 sequences as well. The use of this gene may allow the introduction and amplification of virtually any genetic element in various new cellular environments.

MeSH Terms

Animals Cell Line Clone Cells/enzymology Cricetinae/genetics Cricetulus/genetics Escherichia coli/genetics Fibroblasts/metabolism Genes, Dominant Genetic Vectors Mice/genetics Nucleic Acid Amplification Techniques Nucleic Acid Hybridization Plasmids Tetrahydrofolate Dehydrogenase/genetics Transformation, Genetic

Chemicals

Tetrahydrofolate Dehydrogenase

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Wigler M

Perucho M

Kurtz D

Dana S

Pellicer A

Axel R

Silverstein S

References (23)

23 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.77.6.3567

Published

1980-06-00

Pages

3567-70

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC349658

Subset

External Links

PubMed source DOI: 10.1073/pnas.77.6.3567 PMC Full Text View journal details

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Transformation of mammalian cells with an amplifiable dominant-acting gene.

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