New positive selection system based on the parD (kis/kid) system of the R1 plasmid.
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.
Gabant
P
P
Université Libre de Bruxelles, Gosselies, Belgium. pgabant@dbm.ulb.ac.be
Van Reeth
T
T
Drèze
P L
PL
Faelen
M
M
Szpirer
C
C
Szpirer
J
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004268:DNA-Binding Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D011815:R Factors / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D001427:Bacterial Toxins; C068356:CcdB protein, Plasmid F; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C068353:Kid protein, Bacteria; C068354:Kis protein, Bacteria; C489746:ParD protein, E coli; C489749:ParD protein, Plasmid R1; C078613:parD protein, Bacteria