Table 1.

Bacterial strains and plasmids

Strain or plasmidRelevant characteristicsaSource or reference
P. aeruginosa
 ML5087 ilv-220 thr-9001 leu-9001 met-9011 pur-67 aphA 29
 K1111ML5087 nfxB This study
 K1112ML5087nalB This study
E. coli
 DH5α supE44 ΔlacU16980 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 acrAB+ Bethesda Research Laboratories
 KZM120b ΔacrAB::kan 21
 LBB1201KZM120btolC::Tn10 5
 BL21(DE3)F ompT r B m B ; DE3 is a lambda derivative carrying lacI and T7 RNA polymerase genes under placUV5 control 40
 K113BL21(DE3) carrying the phage T7 lysozyme vector pLysS; Cmr 39
Plasmids
 pAK1900 E. coli-P. aeruginosa shuttle cloning vector carrying plac upstream of MCS; AprCbr R. Sharp, Queen’s University
 pKMJ002pAK1900::nfxB mexCD-oprJ;mexCD-oprJ in orientation opposite to plac 31
 pADD214Mini-D replicon derived from phage D3112; Tcr 3
 pPV1pADD214 derivative carrying a P. aeruginosachromosomal fragment which includesmexAB-oprM 32
 pPV20pAK1900::mexAB-oprM mexAB-oprM in orientation opposite to plac 32
 pT7-6pBR322-based expression vector carrying MCS downstream of the strong gene 10 promoter of phage T7; Apr S. Tabor
 pRSK01pT7-6::mexAB This study
 pPV6pAK1900::mexAB; mexAB in orientation opposite to plac 32
 pRSP01pAK1900::mexAB-OprM This study
 pRK415Low-copy-number, broad-host-range cloning vector carrying MCS downstream of plac; Tcr 13
 pRSP06pRK415::oprJ This study
 pRSP08pRK415::oprM This study
 pRSP15pRK415::nfxB mexCD-oprJ This study
 pRSP17pRK415::mexAB-oprM This study
 pRSP19pRK415::mexAB This study
 pRSP23pAK1900::mexCD; mexCDin orientation opposite to plac This study
 pRSP25pRK415::mexCD This study
 pRSP45pAK1900::nfxB mexCD-oprJ This study
 pRSP46pAK1900::mexCD This study
 pSL1180Superlinker phagemid cloning vectorPharmacia
 pET21-d(+)pBR322-derived C-terminal His-Tag vector carrying a T7lac promoter upstream of a MCS; lacIApr Novagen
 pTK15pET21-d(+)::mexB This study
  • a MCS, multiple cloning site; Apr, ampicillin resistant; Cbr, carbenicillin resistant; Tcr, tetracycline resistant; T7lac, phage T7 promoter followed by a lac operator. Unless otherwise indicated, all genes were cloned in the same orientation as plac of pAK1900 or pRK415.

  • b KZM120 is the same strain as HN818.