Table 1.

Bacterial strains and plasmids used in this study

Strain group and strainGenotype or descriptionaSource or reference
P. aeruginosastrains that lack or overexpress individual pumps or combinations of the pumps
 PAM1020PAO1 prototrophThis study
 PAM1032 nalB(mexAB-oprM is overexpressed)This study
 PAM1033 nfxB (mexCD-oprJ is overexpressed)This study
 PAM1034 nfxC(mexEF-oprN is overexpressed)This study
 K590 met-9011 amiE200 rpsL pvd-9 mexA::Tc 30
 K613 met-9011 amiE200 rpsL pvd-9 oprM::ΩHg 31
 PAM1106 mexA::TcThis study
 PAM1154 oprM::ΩHgThis study
 PAM1177 nfxB oprM::ΩHgThis study
 PAM1187 nfxC oprM::ΩHgThis study
 PAM1360 mexA::Tc ΔmexCD-oprJ::GmThis study
 PAM1409ΔmexCD-oprJ::GmThis study
 PAM1610 nalB ΔmexEF-oprN::ΩHgThis study
 PAM1623ΔmexEF-oprN::ΩHgThis study
 PAM1536 nfxB ΔmexAB-oprM::CmThis study
 PAM1554ΔmexAB-oprM::CmThis study
 PAM1561ΔmexAB-oprM::Cm ΔmexCD-oprJ::GmThis study
 PAM1624ΔmexCD-oprJ::Gm ΔmexEF-oprN::ΩHgThis study
 PAM1625ΔmexAB-oprM::Cm ΔmexEF-oprN::ΩHgThis study
 PAM1626ΔmexAB-oprM::Cm ΔmexEF-oprN::ΩHg ΔmexCD-oprJ::GmThis study
Strains with different levels of mexAB-oprMoperon expression containing target-based mutations
P. aeruginosa
  PAM1548 gyrA(Thr83→Ile)This study
  PAM1324 gyrA(Asp87→Tyr)This study
  PAM1572 nalB gyrA(Thr83→Ile)This study
  PAM1573 nalB gyrA(Thr83→Ile)This study
  PAM1481 nalB gyrA(Asp87→Tyr)This study
  PAM1569 nfxB gyrA(Thr83→Ile)This study
  PAM1482 nfxB gyrA(Asp87→Tyr)This study
  PAM1570 nfxC gyrA(Thr83→Ile)This study
  PAM1491 nfxC gyrA(Asp87→Tyr)This study
  PAM1582 nalB gyrA(Thr83→Ile) parC (Ser87→Leu)This study
  PAM1609 nalB gyrA (Thr83→Ile) parC(Ser87→Leu) gyrA (Asp87→Tyr)This study
  PAM1064 mexA-phoA::TcThis study
  PAM1667 gyrA (Thr83→Ile) parC(Ser87→Leu) mexA-phoA::TcThis study
  PAM1669 gyrA (Thr83→Ile) parC(Ser87→Leu) gyrA (Asp87→Tyr)mexA-phoA::TcThis study
  PAM1665 gyrA (Thr83→Ile)oprM::ΩHgThis study
  PAM1600 gyrA (Thr83→Ile) parC(Ser87→Leu) oprM::ΩHgThis study
  PAM1640 gyrA (Thr83→Ile) parC(Ser87→Leu) gyrA (Asp87→Tyr)oprM::ΩHgThis study
E. coli
  DH5α endA hsdR17 supE44 thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169 deoR80dlacΔ(lacZ)]M15 1
  S17-1 thi pro hsdR recATra 36
Plasmids
 pX1918-GTApr Gmr; contains the selectable Gmr marker downstream from the xylEreporter gene 35
 pNOT19Apr; pUC19 with 10-bp NdeI-NotI adaptor inNdeI site 34
 pMOB3Kmr Cmr sacB oriT 34
 pHP45ΩHgApr, HgCl2 r; derivative of pHP45Ω with Ω interposon containing the mer operon from Tn501 4
 pRS14Tcr; contains a 4.3-kbHindIII fragment carrying the mexAB-oprMoperon with a 4.1-kb internal deletion 37
 pSUP202-mexA-phoA Tcr CbrCmr; pSUP202 carrying the 5′ upstream region ofmexA fused to promoterless phoA geneK. Poole
 pX1918-CmApr Cmr; pX1918GT in whichBamHI fragment with Gmr xylE cassette replaced by BamHI fragment with Cmr from pMOB3This study
 pMOB3-GmKmr Gmr; pMOB3 in which BamHI fragment with Cmr replaced by BamHI fragment with Gmr xylEcassette from pX1918-GTThis study
 pAL219Apr; pNOT19 without SalI site (removed by Klenow treatment and religation)This study
 pAL225Apr; pAL219 with 4.3-kbHindIII (contains ΔmexAB-oprM) from pRS14 in HindIIIThis study
 pAL231Apr Cmr; pAL225 withSalI Cmr fragment from pX1918-Cm inSalI (located in mexAB-oprM region)This study
 pAL232Apr Cmr Gmr; pAL231 with 6.9-kb NotI fragment from pMOB3-Gm withoriT, sacB, and Gmr cloned inNotI located in insert portion by NotI partial digestThis study
 pAL234Apr; pNOT19 with 0.65-kb EcoRI-BamHI PCR fragment with part ofmexE in EcoRI-BamHIThis study
 pAL237Apr; pAL234 with 0.98-kbBamHI-HindIII PCR fragment with part ofoprN in BamHI-HindIIIThis study
 pAL239Apr Hgr; pAL237 with 5.5-kb BamHI fragment with Hgr from pHP45ΩHg in BamHIThis study
 pAL241AprGmr; pAL239 with 6.7-kb NotI fragment from pMOB3 with oriT, sacB, and Gmr inNotI located in vector portion by NotI partial digestThis study
 pAL215Apr; pNOT19 with 0.95-kb EcoRI-BamHI PCR fragment withnfxB and part of mexC and 0.97-kbBamHI-HindIII PCR fragment with part ofoprJ in EcoRI-HindIIIThis study
 pAL217Apr Gmr; pAL215 with 2.4-kb BamHI Gmr fragment from pX1918-GT inBamHIThis study
 pAL224AprCmr Gmr; pAL217 with 5.3-kb NotI fragment from pMOB3 with SacB, oriT, and Cmr in NotI located in vector portion byNotI partial digestThis study
  • a ΩHg, Hg resistance derivative of interposon Ω; Apr, ampicillin resistance; Cmr, chloramphenicol resistance; Gmr, gentamicin resistance; Tcr, tetracycline resistance; Kmr, kanamycin resistance; Cbr, carbenicillin resistance; oriT, origin of transfer from RP4;sacB, sacB locus from Bacillus subtilis.