The Induction of DNA Double-strand Breaks in CHO Cells byPvuII: Kinetics Using Neutral Filter Elution (pH 9·6)

1 January 1990

journal article
research article
Published by Taylor & Francis in International Journal of Radiation Biology

Vol. 57 (5) , 933-938
https://doi.org/10.1080/09553009014551051

Abstract

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.

This publication has 12 references indexed in Scilit:

Induction of chromosomal aberrations in CHO cells by restriction endonucleases: effects of blunt- and cohesive-ended double-strand breaks in cells treated by ‘pellet’ methods
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1989
Cell electroporation is a highly efficient method for introducing restriction endonucleases into cells
Mutation Research Letters, 1989
Linear DNA Elution Dose Response Curves Obtained in CHO Cells with Non-unwinding Filter Elution after Appropriate Selection of the Lysis Conditions
International Journal of Radiation Biology, 1989
The induction of chromosome aberrations by restriction endonucleases that produce blunt-end or cohesive-end double-strand breaks
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1988
Use of Restriction Endonucleases to Study Relationships between DNA Double-strand Breaks, Chromosomal Aberrations and Other End-points in Mammalian Cells
International Journal of Radiation Biology, 1988
Electroporation for the efficient transfection of mammalian cells with DNA
Nucleic Acids Research, 1987
The chromosome-breaking activity of the restriction endonuclease Alu I in CHO cells is independent of the S-phase of the cell cycle
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1985
Induction of sister-chromatid exchanges by restriction endonucleases
Mutation Research Letters, 1985
Enzymatic Restriction of Mammalian Cell DNA Using Pvu II and Bam H1: Evidence for the Double-strand Break Origin of Chromosomal Aberrations
International Journal of Radiation Biology, 1984
X-ray induced DNA double strand break production and repair in mammalian cells as measured by neutral filter elution
Nucleic Acids Research, 1979