TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| E. coli strains | ||
| BW20767/pRL27 | RP4-2-Tc::Mu-1 Kan::Tn7 integrant leu-63::IS10 recA1 zbf-5 creB510 hsdR17 endA1 thi uidA (ΔMluI::pir)/pRL27 | 24 |
| DH5α/λpir | φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 hsdR17 deoR thi-1 supE44 gyrA96 relA1/λpir | 24 |
| P. aeruginosa strains | ||
| PAK | Wild-type P. aeruginosa strain | David Bradley |
| PAK exsA::Ω | PAK with exsA disrupted by insertion of Ω cassette | 12 |
| PAK A44 | PAK mucA::Tn5 mutant isolate | This study |
| PAK A61 | PAK mucA::Tn5 mutant isolate | This study |
| PAK mucA22 | Point mutation (ΔG440) in mucA gene of PAK | This study |
| mucA22 algU::Gm | mucA22 with algU disrupted by insertion of Gm cassette | This study |
| mucA22 algR::Gm | mucA22 with algR disrupted by insertion of Gm cassette | This study |
| PAK algU::Gm | PAK with algU disrupted by insertion of Gm cassette | This study |
| PAK algR::Gm | PAK with algR disrupted by insertion of Gm cassette | This study |
| Plasmids | ||
| pCR2.1-TOPO | Cloning vector for the PCR products | Invitrogen |
| pHW0005 | exoS promoter of PAK fused to promoterless lacZ on pDN19lacZΩ | Unhwan Ha |
| pHW0006 | exoT promoter of PAK fused to promoterless lacZ on pDN19lacZΩ | Unhwan Ha |
| pHW0024 | pscN promoter of PAK fused to promoterless lacZ on pDN19lacZΩ | Unhwan Ha |
| pHW0032 | exsA promoter of PAK fused to promoterless lacZ on pDN19lacZΩ | Unhwan Ha |
| pUCP19 | Shuttle vector between E. coli and P. aeruginosa | 38 |
| pWW020 | mucA gene on pUCP19 driven by algU promoter | This study |
| pWW021 | mucA gene on pUCP19 driven by lac promoter | This study |
| pWW025 | algU gene on pUCP19 driven by lac promoter | This study |
| pMMB67EH | Low-copy-number broad-host-range cloning vector | 13 |
| pWW022 | algR gene on pMMB67EH driven by lac promoter | This study |
| pEX18Tc | Gene replacement vector, Tcr; oriT+sacB+ | 19 |
| pEX18Ap | Gene replacement vector, Apr; oriT+sacB+ | 19 |
| pPS856 | Source of Gmr cassette; Apr Gmr | 19 |
| algU::Gm-pEX18Tc | algU disrupted by insertion of Gmr cassette on pEX18Tc | This study |
| algR::Gm-pEX18Ap | algR disrupted by insertion of Gmr cassette on pEX18Ap | This study |