Thymidine-requiring mutants of Dictyostelium discoideum
| Autor: | R A Deering, G Podgorski |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1984 |
| Předmět: |
biology
Mutant DNA replication Cell Biology Thymidylate Synthase biology.organism_classification Molecular biology Thymidylate synthase Dictyostelium discoideum Nuclear DNA chemistry.chemical_compound Kinetics Biochemistry chemistry Bromodeoxyuridine Mutation biology.protein Centrifugation Density Gradient Dictyostelium Thymidine Molecular Biology DNA Research Article |
| Popis: | Two thymidine auxotrophs of Dictyostelium discoideum were isolated which improve the efficiency of in vivo DNA-specific radiolabeling. Mutant HPS400 lacked detectable thymidylate synthetase activity, required 50 micrograms of thymidine per ml, and incorporated sixfold more [3H]thymidine into nuclear DNA than did a wild-type strain. Either dTMP or exogenously provided DNA also permitted growth of this strain. The second mutant, HPS401, was isolated from HPS400 and also lacked thymidylate synthetase activity, but required only 4 micrograms of thymidine per ml for normal growth and incorporated 55 times more thymidine label than did a control strain. Incorporation of the thymidine analog 5'-bromodeoxyuridine was also markedly increased in the mutants. Catalytic properties of the thymidylate synthetase of D. discoideum investigated in cell extracts were consistent with those observed for this enzyme in other organisms. These strains should facilitate studies of DNA replication and repair in D. discoideum which require short-term labeling, DNA of high specific activity, or elevated levels of substitution in DNA by thymidine analogs. |
| Databáze: | OpenAIRE |
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