The effect of antibiotics on the T4polynucleotide ligase catalyzed template dependent polymerization of oligodeoxythymidylates
| Autor: | Bernd W. Kalisch, Johan H. van de Sande |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
Stereochemistry
Oligonucleotides Biology Coliphages chemistry.chemical_compound Poly dA-dT Ethidium Genetics chemistry.chemical_classification DNA ligase Oligoribonucleotides Oligonucleotide Netropsin Templates Genetic Molecular biology Anti-Bacterial Agents Polynucleotide Ligases Kinetics chemistry Polymerization Polynucleotide Ethidium bromide DNA Thymidine Research Article |
| Zdroj: | Nucleic Acids Research. 6:1881-1894 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/6.5.1881 |
| Popis: | The poly(dA) dependent T4 polynucleotide ligase catalyzed polymerization of oligodeoxythymidylates is dependent upon duplex stability. The antibiotics ethidium bromide, netropsin and Hoechst 33258 stabilize the duplex poly(dA) . P(dT)n (n = 6-10) to thermal denaturation. Ethidium bromide to DNA ratio of 1.25 and netropsin or Hoechst 33258 to DNA ratio of 0.1 the Tm of d(pT) 10 . poly (dA) was increased by 10 degrees and 25 degrees C respectively. The T4 polynucleotide ligase activity was not inhibited under these conditions and temperature optimum of joining of d(pT) 10 . poly(dA) was increased 5 degrees to 10 degrees by the binding of the antibiotics. Duplexes containing shorter oligodeoxythymidylates required lower concentrations of the antibiotics netropsin or Hoechst 33258 to show no inhibition of T4 polynucleotide ligase. The temperature optima of joining the duplexes d(pT)6 . POLY(DA) and d(pT) 8 . poly(dA) were increased by 5 degrees C upon binding of the antibiotics. Polyacrylamide gel analysis of the T4 polynucleotide ligase catalyzed joining of the oligodeoxythymidylates showed that the presence of antibiotics affected the product distribution of the polymerized oligomers. |
| Databáze: | OpenAIRE |
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