Abstract
Restriction endonucleases (REs) induce chromosomal aberrations in living cells indicating that the ultimate lesion for the production of chromosomal aberrations is a DNA double-strand break (Obe et al. 1987; Bryant, this Vol.; Morgan et al., this Vol.). It is still not clear how living cells can uptake REs. Different methods have been used to treat cells with REs (for discussion see Bryant 1988; Morgan et al., this Vol.) and all of them are more or less successful when measured as frequencies of chromosomal aberrations induced by REs. From our own results concerning the induction of chromosomal aberrations with AluI following treatment of pellets of 4 × 106 cells (about 10 µl volume) with 8 µl of a suspension of REs and newborn calf serum, we proposed that REs are internalized via an energy-dependent receptor-mediated mechanism (Johannes et al. 1989; Obe et al. 1989; Vasudev and Obe 1987). Analyses of Bryant and Christie (1989) have shown that the uptake of REs using the “pellet technique” is dependent on the shipping buffer in which the REs are suspended. The authors assume that “this may result from a combination of hypertonic shock and the detergent present in the shipping buffer” (Bryant and Christie 1989). In order to avoid a possible influence on the uptake of AluI by the relatively high concentrations of buffer components we analyzed the chromosome breaking effects of AluI by treatment of monolayers in such a way that the concentrations of the buffer components are considerably lower when compared to our earlier method.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Bryant PE (1988) Use of restriction endonucleases to study relationships between DNA double-strand breaks, chromosomal aberrations and other end-points in mammalian cells. Int J Radiat Biol 54: 869–890
Bryant PE, Christie AF (1989) Induction of chromosomal aberrations in CHO cells by restriction endonucleases: effects of blunt- und cohesive-ended double-strand breaks in cells treated by “pellet” method. Mutat Res 213: 233–241
Fraley R, Straubinger RM, Rule G, Springer EL Papahadjopoulos D (1981) Liposome-mediated delivery of deoxyribonucleic acid to cells: enhanced efficiency of delivery related to lipid composition and incubation conditions. Biochemistry 20: 6978–6987
Galloway SM, Deasy DA, Bean CL, Kraynak AR, Armstrong MJ, Bradley MO (1987) Effects of high osmotic strength on chromosome aberrations, sister-chromatid exchanges and DNA strand breaks, and the relation to toxicity. Mutat Res 189: 15–25
Johannes C, Tuschy S, Lamprecht I, Obe G (1989) The intercellular distributions of chromosomal aberrations induced by the restriction endonucleases AluI and Bam HI in Chinese hamster ovary cells are overdispersed. Biol Zentralbl 108: 445–451
Obe G, Vasudev V, Johannes C (1987) Chromosome aberrations induced by restriction endonucleases. In: Obe G, Basler A (eds) Cytogenetics, basic and applied aspects. Springer, Berlin Heidelberg New York, pp 300–314
Obe G, Besoke R, Schmidt S (1989) Receptor-mediated uptake of AluI by Chinese hamster ovary cells: evidence from experiments with various treatment times and treatment temperatures, and with chloroquine, methylamine and monensin. Biol Zentralbl 108: 439–444
Vasudev V, Obe G (1987) Evidence for a receptor-mediated endocytosis of AluI in Chinese hamsterovary cells. Mutat Res 197: 109–116
Winegar RA, Philips JW, Youngblom JH, Morgan W (1989) Cell electroporation is a highly efficient method for introducing restriction endonucleases into cells. Mutat Res 225: 49–53
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1990 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Johannes, C., Obe, G. (1990). Induction of Chromosomal Aberrations by the Restriction Endonuclease AluI in Chinese Hamster Ovary (CHO) Cells: Influence of Glycerol on Aberration Frequencies. In: Obe, G., Natarajan, A.T. (eds) Chromosomal Aberrations. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-75682-5_9
Download citation
DOI: https://doi.org/10.1007/978-3-642-75682-5_9
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-75684-9
Online ISBN: 978-3-642-75682-5
eBook Packages: Springer Book Archive