TABLE 1.

Strains, plasmids, and primers used in this study

Strain, plasmid, or primerDescription or sequenceSource or reference
Strains
    E. coli
        JM109e14mcrA recA1 endA1 gyrA96 thi-1 hsdR17(rK mK+) supE44 relA1 Δ(lac-proAB) [F′ traD36 proAB lacIqM15]Promega
        SM10FsupE44 thi-1 thr-1 leuB6 lacY1 tonA21 recA::RP4-2-Tc::Mu 19
    P. aeruginosa
        PAO1Nonmucoid P. aeruginosa strain O1D. J. Wozniak
        WFPA1 algD::tet in PAO1 45
        WFPA12 algR::aac1 in PAO1 45
        WFPA203PAO1 amrZΔ::tet attB::arabinose-inducible amrZThis study
        WFPA205PAO1 amrZΔ::tetThis study
        AWOΔpilA in PAO1C. Whitchurch
        WFPA510Wild-type amrZ replaced in WFPA205This study
        WFPA511 amrZ K18A in PAO1This study
        WFPA512 amrZ V20A in PAO1This study
        WFPA513 amrZ R22A in PAO1This study
Plasmids
    pJF2Mini-CTX with pBAD30-based sequence and wild-type amrZThis study
    pAB3pEX18Ap ΔamrZ::tet with flanking sequence to construct deletion 31
    pSW196pBAD30-based vector; EcoRI, PstI, SmaI, NotI, and SacI cloning sitesThis study
    PDJW586Wild-type amrZ in pEX18Ap for use in gene replacement 31
    pPJ155 amrZ K18A in pEX18Ap for use in gene replacementThis study
    pPJ156 amrZ V20A in pEX18Ap for use in gene replacementThis study
    pPJ157 amrZ R22A in pEX18Ap for use in gene replacement 31
    pEX18ApShuttle vector based on pUC18 42
    pEX100TGene replacement vector 33
Primers
    pET15′-CAATTCCCCTCTAGAAAGAATTCTGTTTAACTTTAAGAAGG-3′This study
    pET25′-GGGCTTTGTTAGCAGCCGG-3′This study
    amrZ145′-AACGACCGGTGGTCAGAAGG-3′ 31
    amrZ225′-GTTACTCGCCCATGGTTTGG-3′This study
    amrZ195′-CAAATTCGTCGTTGCTCTGCCCGAGGG-3′ 31
    amrZ185′-CTGACAAATTCGCCGTTCGTCTG-3′This study
    amrZ175′-AGCCGTACCGCTGACGCATTCGTCGTTCGTCTG-3′This study