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Cre-loxP Recombination System for Large Genome Rearrangements in Lactococcus lactis

By Nathalie Campo, Marie-Line Daveran-Mingot, Kees Leenhouts, Paul Ritzenthaler and Pascal Le Bourgeois


We have used a new genetic strategy based on the Cre-loxP recombination system to generate large chromosomal rearrangements in Lactococcus lactis. Two loxP sites were sequentially integrated in inverse order into the chromosome either at random locations by transposition or at fixed points by homologous recombination. The recombination between the two chromosomal loxP sites was highly efficient (approximately 1 × 10(−1)/cell) when the Cre recombinase was provided in trans, and parental- or inverted-type chromosomal structures were isolated after removal of the Cre recombinase. The usefulness of this approach was demonstrated by creating three large inversions of 500, 1,115, and 1,160 kb in size that modified the lactococcal genome organization to different extents. The Cre-loxP recombination system described can potentially be used for other gram-positive bacteria without further modification

Topics: Genetics and Molecular Biology
Publisher: American Society for Microbiology
Year: 2002
DOI identifier: 10.1128/AEM.68.5.2359-2367.2002
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Provided by: PubMed Central
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