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Res Microbiol. 2008 Sep-Oct;159(7-8):545-9. doi: 10.1016/j.resmic.2008.06.004. Epub 2008 Jun 27.

Construction of an improved RP4 (RK2)-based conjugative system.

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Institut Pasteur, Unité Plasticité du Génome Bactérien, CNRS URA 2171, 25 rue du Dr Roux, 75724 Paris, France.


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.

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