Open AccessExpression of abbreviated mouse dihydrofolate reductase genes in cultured hamster cells.
Author(s) -
Charles S. Gasser,
Christian C. Simonsen,
James Schilling,
Robert T. Schimke
Publication year - 1982
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.79.21.6522
Subject(s) - dihydrofolate reductase , chinese hamster ovary cell , gene , biology , complementary dna , intron , microbiology and biotechnology , transfection , hamster , chinese hamster , coding region , genetics , phenotype , dna , cell culture
We have constructed a number of abbreviated dihydrofolate reductase (DHFR) genes by using cloned mouse genomic and cDNA sequences. These genes contain 1.0 kilobase of 5' flanking genomic sequence and varying portions of the 3' non-coding region. Two of the genes contain the first two introns of the DHFR gene; the other three lack introns. Transfection of DHFR-deficient Chinese hamster ovary cells with any of these constructed genes results in cells with the DHFR+ phenotype. Treatment of the transfectants with methotrexate, a folate antagonist, leads to the emergence of methotrexate-resistant colonies which have amplified the transfected genes.