TABLE 1.

Summary of strains and plasmids used

Strain or vectorCharacteristicsSource or reference
Strain
    NovaBlue singles competent cellsendA1 hsdR17 (rK12 mK12+) supE44 thi-1 recA1 gyrA96 relA1 lac F′[proA+B+ lacIqZΔM15::Tn10] (Tetr)Novagen
    E. coli BL21-CodonPlus(DE3)-RPF ompT hsdS(rB mB) dcm+ Tetr gal λ(DE3) endA Hte [argU proL Camr]Novagen
    E. coli B834(DE3)F ompT hsdSB(rB mB) gal dcm met(DE3)Novagen
    P. aeruginosa mPAO1Wild type24
    P. aeruginosa mPAO1 pilF::Tn5Transposon insertion mutant24
Vector
    pSTBLUE-1Blunt-ended cloning acceptor vectorNovagen
    pET28aT7 expression system inducible by IPTGNovagen
    pET28a-pilFPAO1 pilF constructThis work
    pUCP20GmShuttle vector with SmaI-flanked gentamicin cassette inserted into ScaI site within bla10
    pUCP20Gm-pilFPAO1 pilF constructThis work
    pUCP20Gm-pilF C18GCys18 of PilF putative lipidation site replaced by GlyThis work