Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant propertiesReference or origin
P. aeruginosa
 PAO1Prototroph 15
 PAO196PAO1 with Δ(mexAB-oprM)::Gmr-GFPThis study
 PAO200PAO1 with unmarked Δ(mexAB-oprM)This study
 K337a Same as ML5087 (ilv-220 thr-9001 leu-9001 met-9011 pur-67 aphA) 30
E. coli SM10Kmr; mobilizer strain (thi-1 thr leu tonA lacY supE recA::RP4-2Tc::Mu) 6
Plasmidsb
 pAK1900Apr; broad-host-range cloning vector 30
 pEX100TApr; sacB + oriT +; gene replacement vector 35
 pFLPApr; source of yeast Flp recombinase 14
 pUC18Apr; general purpose cloning and expression vector 41
 pUCP21TApr; mobilizable broad-host-range cloning vector 36
 pRSP01Apr; mexA + mexB + oprM + (8.5-kb chromosomal HindIII fragment cloned into pAK1900) 29
 pRSP14Apr; Δ(mexAB-oprM) (pRSP01 with 4.1-kb SacII deletion) 29
 pPS807Apr; Δ(mexAB-oprM) (1.8-kbHindIII-KpnI fragment from pRSP14 cloned between the same sites of pUC18)This study
 pPS809Apr; PCR-amplified 4.5-kb fragment from pPS807 ligated to 1.8-kb blunt-ended Gmr-GFP fragment from pPS858)This study
 pPS858Apr Gmr; source of Gmr-GFP Gmr-conferring fragment flanked by FRT sites 14
 pPS951Apr Gmr; subcloning of a 5.9-kb blunt-ended HindIII-KpnI fragment from pPS809 into the SmaI site of pEX100TThis study
 pPS952Apr; mexA + mexB + oprM + (8.5-kb chromosomal HindIII fragment from pRSP01 cloned into the same site of pUCPT21T)This study
  • a For a detailed description of other K337 derivatives used in this study, see Materials and Methods.

  • b Details on the construction of recombinant plasmids are presented in Materials and Methods.