TABLE 1.
Strains and plasmidsa
| Strain or plasmid | Description and/or genotype | Reference or source |
|---|---|---|
| Strains | ||
| P. aeruginosa | ||
| PAM1275 | PAO1, mexB::ΩHg | O. Lomovskaya |
| PT5 | PAO1 | Laboratory collection |
| PT149 | PT5nfxC (formerly PAO-7H) | 12 |
| PT640 | PT5, mexB::ΩHg, transduced from PAM1275, Hgr | This study |
| PT644 | PT149, mexB::ΩHg, transduced from PAM1275, Hgr | This study |
| PT644ΔF | PT644, ΔmexF, Hgr | This study |
| E. coli | ||
| DH10B | F−araD139 Δ(ara-leu) 7696 galU galK Δ(lac)X74 mcrΔ(mrr-hsdRMS-mcrBC) deoR Φ80dlacZΔM15 endA1 nupG recA1 rpsL | Laboratory collection |
| Plasmids | ||
| pIC20H | Multipurpose cloning vector, Apr | 15 |
| pME3087 | Mobilizable suicide vector for gene exchange in Pseudomonas spp., Tcr | D. Haas |
| pJQ200 | Low-copy-number suicide vector, Gmr | 23 |
| pMMB207 | Broad-host-range cloning/expression vector, Apr | 19 |
| pMMB207Gm | pMMB207 with a HindIII-XmnI fragment of pJQ200 replacing HindIII-DraI fragment, Gmr | This study |
| pSP858 | Source of Gmr-GFP cassette, Apr | 9 |
| pFLP2 | Plasmid carrying FLP recombinase, Apr | 9 |
| pOPNL4 | pUC119 with a 9-kbp fragment from cosmid pOPN4 containing mexE, mexF and oprN genes, Apr | 12 |
| pICN47 | pIC20H with 2-kbp Eco47III fragment from pOPNL4, Apr | This study |
| pICN64 | pICN47 with 1.7-kbp NcoI-BamHI fragment from pOPNL4; contains the mexF gene, Apr | This study |
| pMEBHN131 | pME3087 with 3.7-kbp BamHI-HindIII fragment from pICN64 carrying the mexF gene, Tcr | This study |
| pMEBHN15 | pMEBHN131 deleted of a 1-kbp PstI internal fragment of mexF and insertion of a 1.8-kbp PstI Gmr-GFP cassette from pSP858, Tcr Gmr | This study |
| pMMBN126 | pMMB207Gm with 3.7-kbp EcoRI-HindIII from pICN64 containing mexF gene, Gmr | This study |
| pL2EK | pMMBN126 carrying mexF gene with mutation E349K, Gmr | This study |
| pL4EK | pMMBN126 carrying mexF gene with mutation E417K, Gmr | This study |
| pL4DDAG | pMMBN126 carrying mexF gene with mutations D410A and D411G, Gmr | This study |
| pL10KE | pMMBN126 carrying mexF gene with mutation K951E, Gmr | This study |
a
Ap, ampicillin; Hg, mercuric chloride; Tc, tetracycline; Km, kanamycin; Gm, gentamicin.