Table 1.
Bacterial strains and plasmids.
| Strain or plasmid | Genotype or relevant characteristic | Reference or source |
|---|---|---|
| E. coli strains | ||
| DH5α | fhuA2 lacΔU169 phoA glnV44 Φ80’ lacZΔM15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 | Invitrogen |
| Stellar | F–, endA1, supE44, thi-1, recA1, relA1, gyrA96, phoA, Φ80d lacZΔM15, Δ(lacZYA - argF) U169, Δ(mrr - hsdRMS - mcrBC), ΔmcrA, λ– | Clontech |
| S17-1λpir | Thi pro hsdR recA RP4-2(Tc∷Mu)(Km∷Tn7) | Simon et al. (1983) |
| BTH101 | F-, cya-99, araD139, galE15, galK16, rpsL1, hsdR2, mcrA1, mcrB1 | Euromedex, Karimova et al. (2001) |
| P. aeruginosa | ||
| PAO1 | Wild type | ATCC 15692 Holloway and Morgan (1986) |
| PAO1∷FimL-FLAG | FimL-FLAG allelic exchange | Inclan et al. (2011) |
| PAO1ΔfimV | In-frame deletion of fimV | This study |
| PAO1∷PilG-HIS | PilG-HIS allelic exchange | Bertrand (2010) |
| PAO1∷PilGD58A-HIS | PilGD58A-HIS allelic exchange | Bertrand (2010) |
| PAO1ΔfimLΔfimV | In-frame deletion of fimL, fimV | This study |
| PAO1ΔpilA | In-frame deletion of pilA | Bertrand et al. (2010) |
| PAO1ΔcyaB | In-frame deletion of cyaB | Inclan et al. (2011) |
| PAO1Δvfr | In-frame deletion of vfr | Whitchurch et al. (2005) |
| PAO1ΔpilG | In-frame deletion of pilG | Bertrand et al. (2010) |
| PAO1ΔchpA | In-frame deletion of chpA | Bertrand et al. (2010) |
| PAO1∷FimL-GFP | FimL-GFP allelic exchange | Inclan et al. (2011) |
| PAO1ΔfimV∷FimL-GFP | In-frame deletion of fimV, FimL-GFP allelic exchange | This study |
| PAO1ΔchpA∷FimL-GFP | In-frame deletion of chpA, FimL-GFP allelic exchange | This study |
| PAO1ΔpilG∷FimL-GFP | In-frame deletion of pilG, FimL-GFP allelic exchange | This study |
| PAO1ΔpilJ | In-frame deletion of pilJ | Bertrand et al. (2010) |
| Plasmids | ||
| pEX100T | Allelic exchange suicide plasmid, Apr, Cbr | Tan et al. (1999) |
| pJB100 | pEX100T derivative replacing SmaI site with the SpeI site | Bertrand et al. (2010) |
| pJB128 | Allelic exchange construct, PilG-HA in pJB100, Apr, Cbr | Jacob Bertrand |
| pJB125 | Allelic exchange construct, PilH-HA in pEX100T, Apr, Cbr | Jacob Bertrand |
| pMBAD18G | Broad host range expression vector, pBAD promoter, Gmr | Endoh and Engel (2009) |
| pJB142 | PilG-HA in pMBAD18G, Gmr | Bertrand (2010) |
| pJB100ΔfimV | Allelic exchange construct, ΔfimV in pJB100, Apr, Cbr | This study |
| pYFI043 | Allelic exchange construct, FimL-GFP in pJB100, Apr, Cbr | Inclan et al. (2011) |
| pMBAD18G-GFP | pMBAD18G derivative, C-terminal GFP fusion vector, Gmr | Endoh and Engel (2009) |
| pMBADFimV-GFP | pMBAD-GFP derivative to express FimL-GFP, Gmr | This study |
| PaQa-reporter | PaQa reporter, PaQa-YFP, PrpoD-mKate2, Apr, Cbr | Persat et al. (2015a) |
| mini-CTX-Placp1-lacZ | lacp1 promoter fused to lacZ in miniCTX, tetr | Fulcher et al. (2010) |
| pkT25 | BACTH vector to express N-terminal T25 fused protein, Kmr | Euromedex, Karimova et al. (2001) |
| pkNT25 | BACTH vector to express C-terminal T25 fused protein, Kmr | Euromedex, Karimova et al. (2001) |
| pUT18C | BACTH vector to express N-terminal T18 fused protein, Apr | Euromedex, Karimova et al. (2001) |
| pUT18 | BACTH vector to express C-terminal T18 fused protein, Apr | Euromedex, Karimova et al. (2001) |
| pkT25-zip | T25 fused N-terminal to a leucine zipper | Euromedex, Karimova et al. (1998) |
| pUT18C-zip | T18 fused N-terminal to a leucine zipper | Euromedex, Karimova et al. (1998) |
| pkT25-FimL | BACTH construct to express T25-FimL, Kmr | This study |
| pkNT25-FimL | BACTH construct to express FimL-T25, Kmr | This study |
| pUT18C-FimL | BACTH construct to express T18-FimL, Apr | This study |
| pUT18-FimL | BACTH construct to express FimL-T18, Apr | This study |
| pkT25-FimV | BACTH construct to express T25-FimV, Kmr | This study |
| pUT18C-FimV | BACTH construct to express T18-FimV, Apr | This study |
| pUT18-FimV | BACTH construct to express FimV-T18, Apr | This study |
| pkT25-PilG | BACTH construct to express T25-PilG, Kmr | This study |
| pkNT25-PilG | BACTH construct to express PilG-T25, Kmr | This study |
| pUT18C-PilG | BACTH construct to express T18-PilG, Apr | This study |
| pUT18-PilG | BACTH construct to express PilG-T18, Apr | This study |
| pkT25-PilGD58A | BACTH construct to express T25-PilGD58A, Kmr | This study |
| pkNT25-PilGD58A | BACTH construct to express PilGD58A-T25, Kmr | This study |
| pUT18C-PilGD58A | BACTH construct to express T18-PilGD58A, Apr | This study |
| pUT18-PilGD58A | BACTH construct to express PilGD58A-T18, Apr | This study |
| pkT25-PilH | BACTH construct to express T25-PilG, Kmr | This study |
| pkNT25-PilH | BACTH construct to express PilG-T25, Kmr | This study |
| pUT18C-PilH | BACTH construct to express T18-PilG, Apr | This study |
| pUT18-PilH | BACTH construct to express PilG-T18, Apr | This study |