Estimation of DNA fork displacement rates from isolated mammalian chromosomes |
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Authors: | Jeff Dieden Robert B. Painter Leon N. Kapp |
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Affiliation: | (1) Laboratory of Radiobiology, University of California, 94143 San Francisco, California, USA |
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Abstract: | Chromosomes were isolated from Chinese hamster ovary cells that had been pulse-labeled with 3H-thymidine and bromodeoxyuridine and allowed to proceed to metaphase in the presence of colcemid. The chromosomes were then irradiated with 313 nm light and the length of the pulse-labeled DNA segments was estimated by the distribution of 3H-DNA in alkaline sucrose gradients. The average rate of fork displacement, calculated from this measurement, was 0.9 m/min, which was in excellent agreement with measurements from interphase cells. By the same method, hydroxyurea was shown to retard DNA chain growth by 50–60% in both isolated chromosomes and interphase cells. |
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