Table 1

Strains and plasmids used in this study

Strain or plasmidRelevant characteristic(s) or sequenceSource or reference
P. aeruginosa
    PAO1Wild type24 (J. S. Mattick laboratory)
    PAO1-GFPP. aeruginosa PAO1 tagged with enhanced GFP (EGFP) in a mini-Tn7 construct; Gmr25
    PAO1 pmrF-GFPP. aeruginosa PAO1 transposon mutant ID35399-PA3553 (Washington Genome Center); tagged with EGFP in a mini-Tn7 construct; Gmr26
    PAO1 lasR rhlR-GFPP. aeruginosa lasR rhlR tagged with EGFP in a mini-Tn7 construct; SmrThis study
E. coli
    HB101recA thi pro leu hsdRM; Smr; strain used for maintenance and proliferation of plasmids27
    DH5αF φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rk mk+) phoA supE44 λ thi-1 gyrA96 relA1Invitrogen
    S17-1pro thi recA hsdR (r m+) Tpr Smr Kmr ΩRP4-2-Tc::Mu-Km::Tn728
Plasmids
    pUX-BF13mob+ ori-R6K; helper plasmid providing the Tn7 transposition functions in trans; Ampr29
    pBK-miniTn7(Smr)-gfpDelivery plasmid for miniTn7-PA1/04/03-GFP; Ampr Smr30
    pEX18ApGWGateway compatible gene replacement vector; Sucs Ampr31
    pPS8560.83-kb blunt-ended SacI fragment from pUCGM ligated into the EcoRV site of pPS854; Ampr Gmr32
    pDONR221Gateway donor vector; KmrInvitrogen
    pDONR221-lasRlasR entry clone; Kmr GmrThis study
    pDONR221-rhlRrhlR entry clone; Kmr GmrThis study
    pEX18Ap-lasRlasR knockout vector; Sucs Ampr GmrThis study
    pEX18Ap-rhlRrhlR knockout vector; Sucs Ampr GmrThis study
    pRK600ori-ColE1 RK2-mob+ RK2-tra+; helper plasmid for conjugation; Cmr27