Stimulation of recombination between homologous sequences on plasmid DNA and chromosomal DNA in Escherichia coli by N-acetoxy-2-acetylaminofluorene

C. Luisi-DeLuca, R. D. Porter, W. D. Taylor

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

A plasmid containing a wild-type lac operon and a tetracycline-resistance gene was covalently modified by N-acetoxy-2-acetylaminofluorene and used to transform two series of Lac- Escherichia coli cell types. Each set contained wild-type and repair-deficient mutants. One set of cells contained a lacY mutation and other a deletion of the entire lac operon. Survival and mutagenesis of the plasmid were measured as a function of the N-acetoxy-2-acetylaminofluorene concentration. The results indicate that when no homologous sequences are present in the chromosomal DNA, mutations occur at a low frequency: at 10% survival the frequency was 1-2 x 10-4 mutants per transformant. When homologous sequences, the lacY allele, are present in the chromosomal DNA, Lac- plasmids are found at a high frequency in a recA-dependent, lexA-independent fashion: at 10% survival the frequency was 5-10 x 10-2 mutants per transformant. Southern blot analysis of the restriction enzyme profiles of the resulting plasmid and the host-cell DNA sequences showed recombinational transfer of host sequences to the N-acetoxy-2-acetylaminofluorene-treated plasmid had occurred. When the host chromosomes contained Lac+ homologous sequences no mutants were found, indicating that the results were not caused by error-prone recombination.

Original languageEnglish (US)
Pages (from-to)2831-2835
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number9 I
DOIs
StatePublished - Jan 1 1984

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Acetoxyacetylaminofluorene
Sequence Homology
Genetic Recombination
Plasmids
Escherichia coli
DNA
Lac Operon
Tetracycline Resistance
Restriction Mapping
Mutation
Southern Blotting
Mutagenesis
Chromosomes
Alleles
Genes

All Science Journal Classification (ASJC) codes

  • General

Cite this

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title = "Stimulation of recombination between homologous sequences on plasmid DNA and chromosomal DNA in Escherichia coli by N-acetoxy-2-acetylaminofluorene",
abstract = "A plasmid containing a wild-type lac operon and a tetracycline-resistance gene was covalently modified by N-acetoxy-2-acetylaminofluorene and used to transform two series of Lac- Escherichia coli cell types. Each set contained wild-type and repair-deficient mutants. One set of cells contained a lacY mutation and other a deletion of the entire lac operon. Survival and mutagenesis of the plasmid were measured as a function of the N-acetoxy-2-acetylaminofluorene concentration. The results indicate that when no homologous sequences are present in the chromosomal DNA, mutations occur at a low frequency: at 10{\%} survival the frequency was 1-2 x 10-4 mutants per transformant. When homologous sequences, the lacY allele, are present in the chromosomal DNA, Lac- plasmids are found at a high frequency in a recA-dependent, lexA-independent fashion: at 10{\%} survival the frequency was 5-10 x 10-2 mutants per transformant. Southern blot analysis of the restriction enzyme profiles of the resulting plasmid and the host-cell DNA sequences showed recombinational transfer of host sequences to the N-acetoxy-2-acetylaminofluorene-treated plasmid had occurred. When the host chromosomes contained Lac+ homologous sequences no mutants were found, indicating that the results were not caused by error-prone recombination.",
author = "C. Luisi-DeLuca and Porter, {R. D.} and Taylor, {W. D.}",
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T1 - Stimulation of recombination between homologous sequences on plasmid DNA and chromosomal DNA in Escherichia coli by N-acetoxy-2-acetylaminofluorene

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AU - Porter, R. D.

AU - Taylor, W. D.

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AB - A plasmid containing a wild-type lac operon and a tetracycline-resistance gene was covalently modified by N-acetoxy-2-acetylaminofluorene and used to transform two series of Lac- Escherichia coli cell types. Each set contained wild-type and repair-deficient mutants. One set of cells contained a lacY mutation and other a deletion of the entire lac operon. Survival and mutagenesis of the plasmid were measured as a function of the N-acetoxy-2-acetylaminofluorene concentration. The results indicate that when no homologous sequences are present in the chromosomal DNA, mutations occur at a low frequency: at 10% survival the frequency was 1-2 x 10-4 mutants per transformant. When homologous sequences, the lacY allele, are present in the chromosomal DNA, Lac- plasmids are found at a high frequency in a recA-dependent, lexA-independent fashion: at 10% survival the frequency was 5-10 x 10-2 mutants per transformant. Southern blot analysis of the restriction enzyme profiles of the resulting plasmid and the host-cell DNA sequences showed recombinational transfer of host sequences to the N-acetoxy-2-acetylaminofluorene-treated plasmid had occurred. When the host chromosomes contained Lac+ homologous sequences no mutants were found, indicating that the results were not caused by error-prone recombination.

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