The induction of DNA double-strand breaks in CHO cells by Pvu II: kinetics using neutral filter elution (pH 9.6).
Chinese hamster CHO K1 cells were treated with the restriction endonuclease Pvu II during electroporation and assayed for DNA double-strand breaks (dsb). Dsb were measured by the non-denaturing filter elution technique (pH 9.6) at various times up to 24 h after restriction endonuclease (RE) treatment. The frequency of dsb following electroporation in the presence of 200 units/ml Pvu II increased over the post-treatment incubation period. This was found not to be due to cell or DNA degradation, indicating that Pvu II remains active for at least 24 h inside the cell. We suggest that these kinetics of dsb result from a competition between incision (by Pvu II) and dsb repair.
Costa
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ND
Department of Biology and Preclinical Medicine, University of St Andrews, Scotland, UK.
Bryant
P E
PE
D000818:Animals; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D066298:In Vitro Techniques
D004247:DNA; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific