TABLE 1.
Summary of strains and plasmids used
| Strain or vector | Characteristics | Source or reference |
|---|---|---|
| Strain | ||
| NovaBlue singles competent cells | endA1 hsdR17 (rK12− mK12+) supE44 thi-1 recA1 gyrA96 relA1 lac F′[proA+B+lacIqZΔM15::Tn10] (Tetr) | Novagen |
| E. coli BL21-CodonPlus(DE3)-RP | F−ompT hsdS(rB− mB−) dcm+ Tetrgal λ(DE3) endA Hte [argU proL Camr] | Novagen |
| E. coli B834(DE3) | F−ompT hsdSB(rB− mB−) gal dcm met(DE3) | Novagen |
| P. aeruginosa mPAO1 | Wild type | 24 |
| P. aeruginosa mPAO1 pilF::Tn5 | Transposon insertion mutant | 24 |
| Vector | ||
| pSTBLUE-1 | Blunt-ended cloning acceptor vector | Novagen |
| pET28a | T7 expression system inducible by IPTG | Novagen |
| pET28a-pilF | PAO1 pilF construct | This work |
| pUCP20Gm | Shuttle vector with SmaI-flanked gentamicin cassette inserted into ScaI site within bla | 10 |
| pUCP20Gm-pilF | PAO1 pilF construct | This work |
| pUCP20Gm-pilF C18G | Cys18 of PilF putative lipidation site replaced by Gly | This work |