TABLE 1.

Strains and vectors used

Strain or plasmidRelevant characteristicsSource or reference
E. coli Top10F mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupGInvitrogen
P. aeruginosa strains
    PAKWild typeJ. Boyd
    R364Tn5-B21 insertion in pilT 56
    S34Tn5-B21 insertion in pilU 57
    PAK-BΩΩ insertion mutant in pilB 25
    PAK-NPTetr cassette inserted into pilA 54
    PAK-ΔS2In-frame deletion of pilS 4
    PAK-pilJTetr cassette inserted into pilJ 10
    mPAO1Wild type 21
    30903ISphoA insertion in pilC 21
    50208ISphoA insertion pilD 21
    52103ISphoA insertion in pilM 21
    21902ISlacZ insertion in pilN 21
    41893ISphoA insertion in pilO 21
    55199ISphoA insertion in pilP 21
    44919ISphoA insertion pilQ 21
    6329ISlacZ insertion in pilY1 21
    15562ISlacZ insertion in fimX 21
    41545ISphoA insertion in fimV 21
    52867ISphoA insertion in fliC 21
    2630ISlacZ insertion in xcpY 21
    6177ISlacZ insertion in xcpZ 21
Plasmids
    pUCP20GmShuttle vector with SmaI-flanked Gm cassette inserted into ScaI site in bla 7
    pECFP-N1Source of cfp for fusions to the N terminus of CFPClontech
    pECFP-C1Source of cfp for fusionsClontech
    pEYFP-N1Source of yfp for fusionsClontech
    pUCP20Gm-cfpCFP constructThis study
    pUCP20Gm-yfpYFP constructThis study
    pUCP20Gm-pilT-cfpPilT-CFP fusion constructThis study
    pUCP20Gm-cfp-pilTCFP-PilT fusion constructThis study
    pUCP20Gm-yfp-pilTYFP-PilT fusion constructThis study
    pUCP20Gm-yfp-pilUYFP-PilU fusion constructThis study
    pUCP20Gm-yfp-pilBYFP-PilB fusion constructThis study
    pUCP20Gm-yfp-pilU1-117YFP-PilU deleted after K117This study
    pUCP20Gm-yfp-pilU118-382YFP-PilU deleted up to V118This study
    pUCP20Gm-yfp-pilU1-156YFP-PilU deleted after S156This study
    pUCP20Gm-yfp-pilU1-168YFP-PilU deleted after H168This study
    pUCP20Gm-yfp-pilU1-197YFP-PilU deleted after P197This study
    pUCP20Gm-yfp-pilU1-221YFP-PilU deleted after G221This study
    pUCP20Gm-yfp-pilU1-244YFP-PilU deleted after F244This study
    pUCP20Gm-yfp-pilT1-156YFP-PilT deleted after T156This study
    pUCP20Gm-yfp-pilT1-168YFP-PilT deleted after S168This study
    pUCP20Gm-yfp-pilT1-197YFP-PilT deleted after P197This study
    pUCP20Gm-yfp-pilT1-221YFP-PilT deleted after G221This study
    pUCP20Gm-yfp-N′pilT-C′pilUYFP-N′PilT-C′PilU chimeraThis study
    pUCP20Gm-yfp-N′pilU-C′pilTYFP-N′PilU-C′PilT chimeraThis study