Skip to main content
SpringerLink
Log in

Selection of wild-type revertants from methotrexate-resistant cells containing an altered dihydrofolate reductase

  • Published:
Somatic Cell Genetics

Abstract

A selection system for wild-type revertants from methotrexate-resistant Chinese hamster ovary cells is described. In the absence of exogenous thymidine, cells use the folate metabolic pathway to generate thymidine 5′-monophosphate from deoxyuridine 5′-monophosphate. Thus, in the presence of methotrexate, the incorporation of labeled deoxyuridine into phenotypic wild-type cells is inhibited whereas resistant cells that are cycling incorporate sufficient radioactivity to be killed. Using several suicide cycles, wild-type revertants have been isolated from methotrexate-resistant cells containing a structurally altered dihydrofolate reductase. These revertants possess a wild-type sensitivity to the cytotoxicity of the drug and contain a reductase with similar properties as wild-type enzyme.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature cited

  1. Thompson, L. H., and Baker, R. M. (1973). InMethods in Cell Biology, Vol. 6, (ed) Prescott D. M., (Academic Press, New York), pp. 209–281.

    Google Scholar 

  2. Thompson, L. H., Harkins, J. L., and Stanners, C. P. (1973).Proc. Natl. Acad. Sci. U.S.A. 70:3904–3098.

    PubMed  Google Scholar 

  3. Fenwick, R. G., Jr., and Caskey, C. J. (1975).Cell 5:115–122.

    PubMed  Google Scholar 

  4. Chasin, L. A., Feldman, A., Konstan, M., and Urlaub, G. (1974).Proc. Natl. Acad. Sci. U.S.A. 71:718–722.

    PubMed  Google Scholar 

  5. Sharp, J. D., Capecchi, N. E., and Capecchi, M. R. (1973).Proc. Natl. Acad. Sci. U.S.A. 70:3145–3149.

    PubMed  Google Scholar 

  6. Flintoff, W. F., Davidson, S. V., and Siminovitch, L. (1976).Somat. Cell Genet. 2:245–261.

    PubMed  Google Scholar 

  7. Flintoff, W. F., Spindler, S. M., and Siminovitch, L. (1976).In Vitro 12:749–757.

    PubMed  Google Scholar 

  8. Gupta, R. S., Flintoff, W. F., and Siminovitch, L. (1977).Can. J. Biochem. 55:445–452.

    PubMed  Google Scholar 

  9. Flintoff, W., and Saya, L. (1978).Somat. Cell Genet. 4:143–156.

    PubMed  Google Scholar 

  10. Blakley, R. L. (1969).The Biochemistry of Folic Acid and Related Pteridines (North Holland Publishing Company, Amsterdam) pp. 139–187.

    Google Scholar 

  11. Oliverio, V. T. (1961).Anal. Chem. 33:263–265.

    Google Scholar 

  12. Whitehead, V. M., Perrault, M. M., and Stelcner, S. (1975).Cancer Res. 35:2985–2990.

    PubMed  Google Scholar 

  13. Stanners, C. P., Elicieri, G. L., and Green, H. (1971).Nature (London), New Biol. 230:52–54.

    Google Scholar 

  14. McBurney, M. W., and Whitmore, G. F. (1974).Cell 2:173–182.

    PubMed  Google Scholar 

  15. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951).J. Biol. Chem. 193:265–275.

    PubMed  Google Scholar 

  16. Lo, T. C. Y., and Sanwal, B. D. (1975).J. Biol. Chem. 250:1600–1602.

    PubMed  Google Scholar 

  17. Rosenstraus, M. J., and Chasin, L. A. (1977).Somat. Cell Genet. 3:323–333.

    PubMed  Google Scholar 

  18. Schimke, R. T., Kaufman, R. J., Alt, F. W., and Kellems, R. F. (1978).Science 202:1051–1055.

    PubMed  Google Scholar 

  19. Chasin, L. A., and Urlaub, G. (1979). InBranbury Report 2, (ed.) Hsie, A. W., O'Neill, J. P., and McElheny, U. K. (Cold Spring Harbor Laboratory, Cold Spring Harbor), pp. 201–208.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, University of Western Ontario, N6A 5C1, London, Ontario, Canada

    Wayne F. Flintoff & Mary Weber

Authors
  1. Wayne F. Flintoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mary Weber
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Flintoff, W.F., Weber, M. Selection of wild-type revertants from methotrexate-resistant cells containing an altered dihydrofolate reductase. Somat Cell Mol Genet 6, 517–528 (1980). https://doi.org/10.1007/BF01539153

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01539153

Keywords

Search

Navigation