Expression of a methotrexate-resistant dihydrofolate reductase gene by transformed hematopoietic cells of mice |
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Authors: | M. Bar-Eli H. D. Stang K. E. Mercola M. J. Cline |
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Affiliation: | 1. Division of Hematology-Oncology and the Jonsson Comprehensive Cancer Center, UCLA School of Medicine, 90024, Los Angeles, California
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Abstract: | DNA-mediated gene transfer was used to introduce DNA from a methotrexate-resistant mouse fibroblast cell line into mouse bone marrow cells. This cell line contained a methotrexate-resistant dihydrofolate reductase, active at 10?4 M methotrexate, which was electrophoretically separable from the wild-type mouse enzyme. Transformed hematopoietic cells were returned to irradiated mice and selected in vivo by methotrexate administration. Some recipients of transformed marrow cells expressed the electrophoretically distinct, methotrexate-resistant dihydrofolate reductase in hematopoietic cells. These observations suggest that successful transformation of marrow stem cells to methotrexate resistance is accomplished by insertion of a dihydrofolate reductase gene coding for a mutant enzyme that is highly resistant to methotrexate. |
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