TABLE 1.

Strains of P. aeruginosa and E. coli and plasmids used in this study

Bacterial strain or plasmidGenotype or phenotypeSource or reference
Strains
    P. aeruginosa
        PAO1Wild type11
        PAO6261PAO1 Δanr49
        PAO-MW20PAO1 rpoS::aacC1 Gmr46
        PAO6251PAO1 ΔarcDABC17
        PAO6281PAO1 gacA::Ω-Sp/Sm36
        KS06PAO1 attB::(PPA3309-lacZ)This study
        KS08PAO6261 attB::(PPA3309-lacZ)This study
        KS10PAO-MW20 attB::(PPA3309-lacZ)This study
        KS11PAO1 attB::(mini-CTX-lacZ)This study
        KS15PAO1 attB::(PPA3309-gfp)This study
        KS16PAO1 ΔPA3309::aacC1 GmrThis study
        KS17PAO1 ΔPA3309This study
        KS27PAO1 attB::(mini-CTX2)This study
        KS28KS17 attB::(mini-CTX2)This study
        KS29KS17 attB::(pKS13)This study
        KS31PAO1 rhlR attB::(PPA3309-lacZ)This study
        KS32PAO1 lasR attB::(PPA3309-lacZ)This study
        KS33PAO6281 attB::(PPA3309-lacZ)This study
        KS35PAO1 ΔrelAThis study
        KS37PAO1 ΔrelA attB::(PPA3309-lacZ)This study
        NB015PAO1 ΔPA4352This study
    E. coli
        DH10BF mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 endA1 araD139 Δ(ara leu) 7697 galU galK λ rpsL nupGGibco BRL (Invitrogen)
        S17 λpirpro thi hsdR+ Tpr Smr; chromosome::RP4-2 Tc::Mu-Km::Tn7/λpir9
        SM10thi-1 thr leu tonA lacY supE recA::RP4-2-Tc::Mu (Kmr)9
    Plasmids
        pEX18ApApr oriT+ sacB+; gene replacement vector with MCS from pUC1823
        Mini-CTX-lacZTcr; promoterless lacZ gene2
        Mini-CTX2Tcr; integration vector24
        pSB219.9AGmr; lasR::Gm suicide vector1
        pSB224.10ATcr; rhlR::Tc suicide vector1
        pPS858Apr Gmr; source of gentamicin cassette23
        pFLP2Apr; source of FLP recombinase23
        pMH305Apr Gmr; source of gfp cassetteMorten Hentzer
        pKS08Apr Gmr; pMH305 with 526 bp of PA3309 promoter between EcoRI and BamHIThis study
        pKS09Tcr; mini-CTX2 with NotI fragment liberated from pKS08 containing PA3309 promoter fused to gfpThis study
        pKS10Apr Gmr; pEX18Ap with 723-bp promoter of PA3309, Gmr-gfp fragment from pPS858 and 625 bp downstream of coding region of PA3309 between EcoRI and SalIThis study
        pKS13Tcr; mini-CTX2 with 1,200-bp PCR fragment covering entire PA3309 gene and 455 bp of putative promoter region and 289 bp of downstream region between EcoRI and HindIIIThis study
        pKS15Tcr; mini-CTX-lacZ containing 492-bp fragment of putative promoter region of PA3309 gene between EcoRI and BamHIThis study
        pKS18Apr Gmr; pEX18Ap with 796-bp promoter of relA (PA0934), Gmr-gfp fragment from pPS858, and 824 bp of the 3′ coding region of relA between EcoRI and HindIIIThis study
        pNB007Apr Gmr; pEX18Ap with 596-bp promoter PA4352, Gmr-gfp fragment from pPS858, and 617 bp downstream of coding region of PA4352 between SacI and HindIIIThis study