Transfection of a nonactive site mutant murine DHFR cDNA (the tryptophan 15 mutant) into Chinese hamster ovary and mouse marrow progenitor cells imparts MTX resistance in vitro.

D. Banerjee, S. C. Zhao, Y. Tong, J. Steinherz, Kira Gritsman, J. R. Bertino

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Chinese hamster ovary (CHO) DHFR-cells were converted to the DHFR+ (dihydrofolate reductase) phenotype when transfected with a mammalian expression vector containing the murine mutant Trp15 DHFR cDNA. Transfection of the Trp15 DHFR cDNA into wild-type CHO cells resulted in resistance to high levels of methotrexate (MTX) in vitro indicating that this mutant DHFR cDNA can act as a dominant marker. Southern and Northern blot analyses of transfected cells indicated that the transfected mutant DHFR cDNA was integrated and expressed. Gene copy number analysis showed that the Trp15 cDNA was amplifiable in increasing concentrations of MTX. Transfection of murine bone marrow progenitor cells with the Trp15 mutant DHFR cDNA resulted in MTX resistant colony forming units-granulocyte macrophage.

Original languageEnglish (US)
Pages (from-to)181-184
Number of pages4
JournalCancer Gene Therapy
Volume1
Issue number3
StatePublished - Sep 1994
Externally publishedYes

Fingerprint

Cricetulus
Methotrexate
Tryptophan
Transfection
Ovary
Stem Cells
Complementary DNA
Bone Marrow
Tetrahydrofolate Dehydrogenase
Granulocyte-Macrophage Progenitor Cells
Gene Dosage
Southern Blotting
Bone Marrow Cells
Northern Blotting
In Vitro Techniques
Phenotype

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

Cite this

Transfection of a nonactive site mutant murine DHFR cDNA (the tryptophan 15 mutant) into Chinese hamster ovary and mouse marrow progenitor cells imparts MTX resistance in vitro. / Banerjee, D.; Zhao, S. C.; Tong, Y.; Steinherz, J.; Gritsman, Kira; Bertino, J. R.

In: Cancer Gene Therapy, Vol. 1, No. 3, 09.1994, p. 181-184.

Research output: Contribution to journalArticle

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abstract = "Chinese hamster ovary (CHO) DHFR-cells were converted to the DHFR+ (dihydrofolate reductase) phenotype when transfected with a mammalian expression vector containing the murine mutant Trp15 DHFR cDNA. Transfection of the Trp15 DHFR cDNA into wild-type CHO cells resulted in resistance to high levels of methotrexate (MTX) in vitro indicating that this mutant DHFR cDNA can act as a dominant marker. Southern and Northern blot analyses of transfected cells indicated that the transfected mutant DHFR cDNA was integrated and expressed. Gene copy number analysis showed that the Trp15 cDNA was amplifiable in increasing concentrations of MTX. Transfection of murine bone marrow progenitor cells with the Trp15 mutant DHFR cDNA resulted in MTX resistant colony forming units-granulocyte macrophage.",
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