Expression of prokaryotic genes for hygromycin B and G418 resistance as dominant-selection markers in mouse L cells
| Autor: | J. N. Hobbs, Robert J. Schmidt, Robert Frank Santerre, Norris E. Allen, R.Nagaraja Rao |
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| Rok vydání: | 1984 |
| Předmět: |
R Factors
Biology Molecular cloning medicine.disease_cause Microbiology Mice chemistry.chemical_compound L Cells Transformation Genetic Plasmid Acetyltransferases Escherichia coli Genetics medicine Animals Cloning Molecular Gene Southern blot Kanamycin Kinase Phosphotransferases Genetic transfer Drug Resistance Microbial General Medicine Molecular biology Transformation (genetics) chemistry Genes Bacterial Hygromycin B Plasmids |
| Zdroj: | Gene. 30:147-156 |
| ISSN: | 0378-1119 |
| Popis: | Novel bacterial resistance genes were cloned and expressed as dominant selection markers in mammalian cells. Escherichia coli genes coding for resistance to the aminocyclitol antibiotics hygromycin B (Hm) and G418 were cloned into the eukaryotic expression plasmid pSV5GPT [Mulligan and Berg, Proc. Natl. Acad. Sci. USA 78 (1981) 2072-2076]. Mouse cells normally sensitive to 100 micrograms/ml Hm were transformed with these plasmids and selected in 200 micrograms/ml Hm. Transformants resistant to as much as 1 mg/ml Hm and 500 micrograms/ml G418 were isolated. Cell extracts contained an acetyltransferase activity capable of acetylating G418 and an Hm aminocyclitol phosphotransferase activity. Plasmid DNA sequences were identified by Southern blot analysis of high Mr DNA isolated from transformed cells. |
| Databáze: | OpenAIRE |
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