PMID: 11336980May 5, 2001Paper

In vitro replication and mutagenesis of a novel reversion vector with selective DNA damage in the supF gene

Mutation Research
N M KingK Dixon

Abstract

The ability to detect the most common type of UV-induced mutation, the C to T transition, at the previously characterized hotspot at position 99 of the supF gene has been demonstrated in a selectively irradiated reversion vector, pLS189(Rev). The supF region was amplified, irradiated with 500J/m(2) UVC or unirradiated, and ligated into the pLS189(Rev) plasmid. A portion of ligated product plasmid containing the irradiated fragment was sensitive to nicking by T4 endonuclease V, indicating the presence of the most common type of UV-induced damage, the pyrimidine dimer. Plasmid containing the irradiated or unirradiated supF gene was replicated completely in cellular extracts from either HeLa or XP-A cells in vitro. Plasmid containing the irradiated supF gene showed an inhibition of total replication to a level similar to those of previous studies with plasmid molecules exposed in their entirety to 40J/m(2). Replication of selectively irradiated plasmid resulted in an average reversion frequency of 0.071% in the two extracts; a 42-fold increase over the average spontaneous reversion frequency of unirradiated plasmid. The reversion frequencies were not significantly different between extracts prepared from HeLa and XP-A cells, indic...Continue Reading

References

Nov 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·J J Li, T J Kelly
Aug 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·D C Thomas, T A Kunkel
Apr 19, 1996·The Journal of Biological Chemistry·M P CartyK Dixon
Jun 14, 1996·Science·J R NelsonD C Hinkle
Jan 7, 1997·Proceedings of the National Academy of Sciences of the United States of America·K H Kraemer
Jun 6, 2000·Mutation Research·K A Canella, M M Seidman

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Citations

Sep 20, 2008·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Ulf-Peter GuentherMarkus Schuelke

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