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Illegitimate recombination in an Escherichia coli plasmid: modulation by DNA damage and a new bacterial gene.

Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.

ABSTRACT

We studied DNA rearrangements in Escherichia coli by using a plasmid-based system with a transcriptionally silent tet gene and selecting for Tetr isolates. The predominant activating event was a 1.3-kilobase-pair deletion in the plasmid between two sites, with 14 of 19 base pairs being identical. These deletions occurred equally frequently in a recA+ strain and a recA13 mutant. However, the frequency of Tetr occurrence was stimulated 50-fold by treatment of the cells with UV light in a process that was at least partly independent of the SOS response. Bacterial mutants deleted for the xth-pnc region of the chromosome exhibited a strongly elevated spontaneous frequency of Tetr isolates, all with the same 1.3-kilobase-pair deletion. This phenotype of high-frequency deletion could be complemented by an episome covering this region, but not by the cloned xth gene. These studies helped to define the role of different DNA damages in illegitimate recombination and identify a region of the E. coli chromosome that contains a gene whose product normally suppresses illegitimate deletions.

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