Construction of an improved RP4 (RK2)-based conjugative system.
In many bacteria, gene replacement can only be achieved using RP4-based conjugative systems where exogenous DNA is delivered to the recipient on conditionally replicating plasmids carrying an oriT(RP4). In the commonly used strains SM10 and S17-1 conjugative functions are provided by an RP4 plasmid inserted in the chromosome. Those strains, besides mobilizing oriT(RP4)-carrying plasmids, also transfer their own chromosomal genes, which can be inconvenient for several reasons. We describe here a new version of an RP4-based conjugative system that allows for efficient transfer of mobilizable plasmids without the transfer of chromosomal genes, providing fully controlled and efficient exogeneous DNA delivery to non-transformable bacteria.
Babic
Ana
A
Institut Pasteur, Unité Plasticité du Génome Bactérien, CNRS URA 2171, 25 rue du Dr Roux, 75724 Paris, France. ababic@pasteur.fr
Guérout
Anne-Marie
AM
Mazel
Didier
D
D001419:Bacteria / Q000235:genetics*; D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D018741:Replication Origin
D004269:DNA, Bacterial