Apparent and real recombination frequencies in multicopy plasmids: The need for a novel approach in frequency determination

Frederic Chedin, Rozenn Dervyn, S. Dusko Ehrlich, Philippe Noirot

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Recombination studies of bacteria are often carried out with multicopy plasmids, and recombination frequencies are often deduced from the proportion of cells in the population that express a recombinant phenotype. These frequencies should however be called apparent frequencies, since detection of the recombinant cells requires not only the formation of a rearranged plasmid but also its establishment in the cell. The establishment of the recombinant plasmid can possibly be affected by its interaction with the parental plasmids. To test this hypothesis, we have used a plasmid system enabling the study of deletion formation between short direct repeats (18 bp) in Bacillus subtilis and developed a method by which deletion frequencies are measured under conditions under which interaction is abolished. Real deletion frequencies were thus determined and compared with apparent deletion frequencies. Real frequencies were underestimated by a factor ranging from 4- to 500-fold, depending upon the plasmid under study. This implies that a large majority of the recombinant molecules that are formed are generally not detected. We show that apparent deletion frequencies strongly depend upon (i) the parental plasmid copy number, (ii) the ability of the recombinant molecules to form heterodimeric plasmids, and (iii) the fitness of the recombinant molecules relative to that of parental molecules. Finally, we show that under conditions under which all recombinant molecules are scored, transcription can inhibit the deletion process 10-fold.

Original languageEnglish (US)
Pages (from-to)754-761
Number of pages8
JournalJournal of Bacteriology
Volume179
Issue number3
DOIs
StatePublished - Jan 1 1997

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Genetic Recombination
Plasmids
Nucleic Acid Repetitive Sequences
Bacillus subtilis
Bacteria
Phenotype
Population

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Apparent and real recombination frequencies in multicopy plasmids : The need for a novel approach in frequency determination. / Chedin, Frederic; Dervyn, Rozenn; Ehrlich, S. Dusko; Noirot, Philippe.

In: Journal of Bacteriology, Vol. 179, No. 3, 01.01.1997, p. 754-761.

Research output: Contribution to journalArticle

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