Characterization of large mammalian DNA species sedimented in a reorienting zonal rotor
| Autor: | K.T. Wheeler, J.D. Linn, R. Franklin, E.L. Pautler |
|---|---|
| Rok vydání: | 1975 |
| Předmět: |
DNA damage
Biophysics DNA Single-Stranded Biochemistry Nitrosourea Compounds Choline chemistry.chemical_compound Leucine Methods Animals Molecule Trypsin Molecular Biology chemistry.chemical_classification Isotope Brain Neoplasms Chemistry DNA Neoplasm Cell Biology Centrifugation Zonal Rats Enzyme Covalent bond Nucleic acid Tritium DNA |
| Zdroj: | Analytical Biochemistry. 64:329-342 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/0003-2697(75)90440-6 |
| Popis: | The characteristics of a Beckman-designed slow acceleration unit for the reorientation of alkaline sucrose gradients in a Ti-15 zonal rotor are described. The large DNA species (> 250S) obtained from cultured rat brain tumor cells with this system sediment linearly with time, have virtually no [3H]leucinelabeled or covalently bonded [3H]choline-labeled material sedimenting with them, sediment independently of smaller single-stranded DNA molecules (⋍ 165S) and are 60–80% degraded by the single-strand-specific S1 nuclease. Therefore, it is postulated that these species are collapsed, partially denatured DNA molecules or a collapsed form of single-stranded DNA. When cells were labeled with [14C]TdR, then frozen and stored at − 79°C, this system could detect radiation-induced DNA damage from decay of the incorporated label at accumulated doses as small as 18–126 rads. |
| Databáze: | OpenAIRE |
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