Antibiotic marker modifications of lambda Red and FLP helper plasmids, pKD46 and pCP20, for inactivation of chromosomal genes using PCR products in multidrug-resistant strains.
The Red recombinase system of bacteriophage Lambda has been used to inactivate chromosomal genes in bacteria using PCR products. In this study, we describe the replacement of the ampicillin resistance marker of helper plasmids pKD46 and pCP20 by a gentamicin resistance gene to disrupt chromosomal genes and then to eliminate FRT flanked resistance gene in multiple antibiotic-resistant Salmonella enterica strains.
Doublet
Benoît
B
INRA, UR1282, Infectiologie Animale et Santé Publique, Nouzilly, 37380, France. Benoit.Doublet@tours.inra.fr
Douard
Gregory
G
Targant
Hayette
H
Meunier
Danièle
D
Madec
Jean-Yves
JY
Cloeckaert
Axel
A
D000667:Ampicillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D010582:Bacteriophage lambda / Q000201:enzymology* / Q000235:genetics; D002876:Chromosomes, Bacterial / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D017353:Gene Deletion; D044404:Genomic Islands / Q000235:genetics; D005839:Gentamicins / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D045522:Recombinases / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000235:genetics / Q000821:virology
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D045522:Recombinases; D000667:Ampicillin; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase