Somatic Cell and Molecular Genetics

, Volume 13, Issue 1, pp 67–76 | Cite as

Expression of mouse dihydrofolate reductase gene confers methotrexate resistance in transgenic petunia plants

  • David A. Eichholtz
  • Stephen G. Rogers
  • Robert B. Horsch
  • Harry J. Klee
  • Maria Hayford
  • Nancy L. Hoffmann
  • Sarah B. Braford
  • Cindy Fink
  • Jeff Flick
  • Keith M. O'Connell
  • Robert T. Fraley


Transgenic petunia plants containing an altered (Leu22→Arg22) mouse dihydrofolate reductase gene fused to the cauliflower mosiac virus 35S (CaMV 35S) promoter and nopaline synthase (nos) polyadenylation site were obtained by transforming petunia leaf disks with anAgrobacterium tumefaciens strain carrying the chimeric gene. Transformants were directly selected for and rooted on medium containing 1ΜM methotrexate (MTX). The chimeric gene was present in the regenerated plants at one to three copies and produced the expected 950-nucleotide-long transcript based on Southern and Northern hybridization analyses, respectively. Leaf pieces from the regenerated transgenic plants were able to form callus when cultured on medium containing 1 ΜM MTX and were able to incorporate32P into high-molecular-weight DNA in the presence of > 100 ΜM MTX, thus demonstrating that the chimeric mouse dhfr gene was fully functional and useful as a selectable marker in plant transformation experiments. To date, this is the first report of successful expression of a vertebrate gene in transformed plant cells.


Methotrexate Leaf Disk Chimeric Gene Northern Hybridization Chimeric Mouse 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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Copyright information

© Plenum Publishing Corporation 1987

Authors and Affiliations

  • David A. Eichholtz
    • 1
  • Stephen G. Rogers
    • 1
  • Robert B. Horsch
    • 1
  • Harry J. Klee
    • 1
  • Maria Hayford
    • 1
  • Nancy L. Hoffmann
    • 1
  • Sarah B. Braford
    • 1
  • Cindy Fink
    • 1
  • Jeff Flick
    • 1
  • Keith M. O'Connell
    • 1
  • Robert T. Fraley
    • 1
  1. 1.Monsanto CompanySt. Louis

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