TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant properties | Reference or origin |
|---|---|---|
| P. aeruginosa | ||
| PAO1 | Prototroph | 15 |
| PAO196 | PAO1 with Δ(mexAB-oprM)::Gmr-GFP | This study |
| PAO200 | PAO1 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 SM10 | Kmr; mobilizer strain (thi-1 thr leu tonA lacY supE recA::RP4-2Tc::Mu) | 6 |
| Plasmidsb | ||
| pAK1900 | Apr; broad-host-range cloning vector | 30 |
| pEX100T | Apr; sacB+oriT+; gene replacement vector | 35 |
| pFLP | Apr; source of yeast Flp recombinase | 14 |
| pUC18 | Apr; general purpose cloning and expression vector | 41 |
| pUCP21T | Apr; mobilizable broad-host-range cloning vector | 36 |
| pRSP01 | Apr; mexA+mexB+oprM+ (8.5-kb chromosomal HindIII fragment cloned into pAK1900) | 29 |
| pRSP14 | Apr; Δ(mexAB-oprM) (pRSP01 with 4.1-kb SacII deletion) | 29 |
| pPS807 | Apr; Δ(mexAB-oprM) (1.8-kb HindIII-KpnI fragment from pRSP14 cloned between the same sites of pUC18) | This study |
| pPS809 | Apr; PCR-amplified 4.5-kb fragment from pPS807 ligated to 1.8-kb blunt-ended Gmr-GFP fragment from pPS858) | This study |
| pPS858 | Apr Gmr; source of Gmr-GFP Gmr-conferring fragment flanked by FRT sites | 14 |
| pPS951 | Apr Gmr; subcloning of a 5.9-kb blunt-ended HindIII-KpnI fragment from pPS809 into the SmaI site of pEX100T | This study |
| pPS952 | Apr; 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.