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PMID: 2684636 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Retracted Publication

Breakage--reunion and copy choice mechanisms of recombination between short homologous sequences.

The EMBO journal ·Vol. 8 ·No. 10 ·1989-10-00 ·Pages 3127-33

Brunier D, Peeters BP, Bron S, Ehrlich SD

Abstract

To study recombination between short homologous sequences in Escherichia coli we constructed plasmids composed of the pBR322 replicon, M13 replication origin and a recombination unit inserted within and inactivating a gene encoding chloramphenicol resistance. The unit was composed of short direct repeats (9, 18 or 27 bp) which flanked inverted repeats (0, 8 or 308 bp) and a gene encoding kanamycin resistance. Recombination between direct repeats restored a functional chloramphenicol resistance gene, and could be detected by a simple phenotype test. The plasmids replicated in a double-stranded form, using the pBR322 replicon, and generated single-stranded DNA when the M13 replication origin was activated. The frequency of chloramphenicol-resistant cells was low (10(-8)-10(-4] when no single-stranded DNA was synthesized but increased greatly (to 100%) after induction of single-stranded DNA synthesis. Recombination between 9 bp direct repeats entailed no transfer of DNA from parental to recombinant plasmids, whereas recombination between 18 or 27 bp repeats entailed massive transfer. The presence or length of inverted repeats did not alter the pattern of DNA transfer. From these results we propose that direct repeats of 9 bp recombine by a copy choice process, while those greater than or equal to 18 bp can recombine by a breakage-reunion process. Genome rearrangements detected in many organisms often occur by recombination between sequences less than 18 bp, which suggests that they may result from copy choice recombination.

MeSH Terms
Base Sequence Chloramphenicol Resistance/genetics DNA, Single-Stranded/genetics Escherichia coli/genetics Genes, Bacterial Genotype Plasmids Recombination, Genetic Repetitive Sequences, Nucleic Acid Replicon Sequence Homology, Nucleic Acid Transformation, Bacterial
Chemicals
DNA, Single-Stranded
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Brunier D
Laboratoire de Génétique Microbienne, Institut de Biotechnologie, Jouy en Josas, France.
Peeters B P
Bron S
Ehrlich S D
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1989-10-00
Pages
3127-33
Language
English
Region
England
NLM ID
8208664
PMCID
PMC401393
Subset
IM
Corrections
RetractionIn
PMID: 2203648
External Links
PubMed source PMC Full Text View journal details
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