TABLE 1.

Plasmids and bacterial strains used in this study

Strain or plasmidRelevant characteristics or purposeaReference or source
P. aeruginosa strains
    PA14Wild-type laboratory strain; Rifr47
    lasR rhlR mutantPA14 with in-frame deletion of lasR rhlR; Rifr46
    mvfR mutantPA14 with TnphoA insertion at mvfR; Rifr Kmr4
    rpoS mutantPA14 with in-frame deletion of rpoS; Rifr46
    oxyR mutantPA14 with in-frame deletion of oxyR; Rifr12
    phoB mutantPA14 with in-frame deletion of phoB; RifrThis study
    S33N mutantoxyR with the chromosomally integrated mTn7-oxyR(S33N); Rifr GmrThis study
    C25S mutantoxyR with the chromosomally integrated mTn7-oxyR(C25S); Rifr GmrThis study
    C199A mutantoxyR with the chromosomally integrated mTn7-oxyR(C199A); Rifr GmrThis study
    C199S mutantoxyR with the chromosomally integrated mTn7-oxyR(C199S); Rifr GmrThis study
    C208S mutantoxyR with the chromosomally integrated mTn7-oxyR(C208S); Rifr GmrThis study
    C199A C208S mutantoxyR with the chromosomally integrated mTn7-oxyR(C199A C208S); Rifr GmrThis study
    C199S C208S mutantoxyR with the chromosomally integrated mTn7-oxyR(C199S C208S); Rifr GmrThis study
E. coli strains
    DH5αMultipurpose cloning48
    S17-1Conjugal transfer of mobilizable plasmids; Tpr Smr53
    BL21(DE3)/pLysSHigh-stringency T7 promoter-based gene expressionNovagen
Plasmids
    pQF50lacZ transcriptional fusion; Cbr16
    pQF-N10pQF50 with the katA promoter N10 (−133 to +164); CbrThis study
    pQF-N21pQF50 with the katA promoter N21 (−76 to +164); CbrThis study
    pQF-N22pQF50 with the katA promoter N22 (−56 to +164); CbrThis study
    pQF-N23pQF50 with the katA promoter N23 (−35 to +164); CbrThis study
    pUCP18General-purpose cloning; Cbr50
    pUCP18-oxyR-FLAGpUCP18 with the wild-type oxyR gene, with C-terminal FLAG taggingc; CbrThis study
    pUCP18-oxyR(C199S)-FLAGpUCP18 with the oxyR C199S mutant gene, with C-terminal FLAG taggingc; CbrThis study
    pUCP18-oxyR(C208S)-FLAGpUCP18 with the oxyR C208S mutant gene, with C-terminal FLAG taggingc; CbrThis study
    pUCP18-oxyR(C199S C208S)-FLAGpUCP18 with the oxyR C199S C208S mutant genes, with C-terminal FLAG taggingc; CbrThis study
    pUC18T-mini-Tn7-Gmmini-Tn7-Gm for single-copy complementation mobilizable from S17-1; Cbr Gmr10
    pTNS2Transposase for mini-Tn7; Cbr10
    mTn7-oxyRpUC18T-mini-Tn7-Gm with the wild-type oxyR gene; Cbr GmrThis study
    mTn7-oxyR(S33N)pUC18T-mini-Tn7-Gm with the oxyR S33N mutant gene; Cbr GmrThis study
    mTn7-oxyR(C25S)pUC18T-mini-Tn7-Gm with the oxyR C25S mutant gene; Cbr GmrThis study
    mTn7-oxyR(C199A)pUC18T-mini-Tn7-Gm with the oxyR C199A mutant gene; Cbr GmrThis study
    mTn7-oxyR(C199S)pUC18T-mini-Tn7-Gm with the oxyR C199S mutant gene; Cbr GmrThis study
    mTn7-oxyR(C208S)pUC18T-mini-Tn7-Gm with the oxyR C208S mutant gene; Cbr GmrThis study
    mTn7-oxyR(C199AC208S)pUC18T-mini-Tn7-Gm with the oxyR C199A and C208S mutant genes; Cbr GmrThis study
    mTn7-oxyR(C199SC208S)pUC18T-mini-Tn7-Gm with the oxyR C199S and C208S mutant genes; Cbr GmrThis study
    pET15bT7 promoter-based expression vector; CbrNovagen
    pET15H-oxyRpET15b with the oxyR gene, with direct His taggingb; CbrThis study
  • a Rifr, rifampin resistant; Gmr, gentamicin resistant; Cbr, carbenicillin and ampicillin resistant; Tpr, trimethoprim resistant; Smr, streptomycin resistant.

  • b Hexa-His tagging directly to the initiation codon, which is derived from the oligonucleotide primer (see Table S1 in the supplemental material).

  • c FLAG tagging directly to the last codon of the OxyR protein, which is derived from the oligonucleotide primer (see Table S1 in the supplemental material).