Cre-loxP recombination system for large genome rearrangements in Lactococcus lactis.
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 x 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.
Campo
Nathalie
N
Laboratoire de Microbiologie et Génétique Moléculaire du CNRS, UMR5100, Université Paul Sabatier, 31062 Toulouse, France.
Daveran-Mingot
Marie-Line
ML
Leenhouts
Kees
K
Ritzenthaler
Paul
P
Le Bourgeois
Pascal
P
D007446:Chromosome Inversion; D002876:Chromosomes, Bacterial; D003470:Culture Media; D015321:Gene Rearrangement; D016680:Genome, Bacterial; D013294:Lactococcus lactis / Q000235:genetics* / Q000254:growth & development; D008970:Molecular Weight; D011995:Recombination, Genetic
D003470:Culture Media