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New positive selection system based on the parD (kis/kid) system of the R1 plasmid

par Gabant, Philippe ;Van Reeth, Thierry ;Dreze, Pierre Luc ;Faelen, Michel ;Szpirer, Claude ;Szpirer, Josiane
Référence BioTechniques, 28, page (734-788)
Publication Publié, 2000

Article révisé par les pairs

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Résumé :

The use of vectors that are designed to allow positive selection of recombinants facilitates cloning experiments in E. coli. Using kid, a lethal gene of the R1 plasmid parD locus, we generated pKID vectors leading to high selective efficiency of recombinants (greater than 90%). The E. coli bacterial host used to propagate these vectors produces the Kis protein, the natural antagonist of Kid. This new positive-selection system exhibits the same efficiency as the original ccdB-based selection vectors, pKIL (4). We also show that the ccdB and kid systems are independent. This property increases the potential of plasmidic poison-antidote systems for genetic applications and opens the door to a generation of new vectors containing the two selection systems.