TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Source |
|---|---|---|
| E. coli | ||
| DH5α | F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | CWBIO |
| BL21 | F− ompT hsdS(rBB− mB−) gal dcm (DE3) | CWBIO |
| S17-1 | RP4-2 Tc::Mu Km::Tn7 Tpr Smr Pro Res− Mod+ | R. Ramphal |
| S17/pEX18Tc-Δvfr | S17-1 harboring the pEX18Tc-Δvfr vector; Tcr | 58 |
| BL21/pETMAL-osaR | BL21 harboring the pETMAL-osaR vector; Kanr | 20 |
| P. aeruginosa | ||
| PAK | Wild type | David Bradley |
| PAK19 | PAK harboring the pUCP19 vector; Cbr | This study |
| ΔosaR | PAK with osaR gene deleted | 20 |
| ΔosaR19 | ΔosaR harboring the pUCP19 vector; Cbr | This study |
| Δvfr | PAK with vfr gene deleted | 58 |
| Δvfr19 | Δvfr harboring the pUCP19 vector; Cbr | This study |
| ΔosaRΔvfr | PAK with both osaR and vfr genes deleted | This study |
| ΔosaRΔvfr19 | ΔosaRΔvfr harboring the pUCP19 vector; Cbr | This study |
| osaROE | PAK harboring the pUCP19-osaR vector for overexpression of osaR; Cbr | 20 |
| vfrOE | PAK harboring the pKF912 vector for overexpression of vfr; Cbr | This study |
| ΔosaRΔvfr/osaR | ΔosaRΔvfr mutant harboring the pUCP19-osaR vector for complement of osaR; Cbr | This study |
| ΔosaRΔvfr/vfr | ΔosaRΔvfr mutant harboring the pKF912 vector for complement of vfr; Cbr | This study |
| Plasmids | ||
| pEX18Tc-Δvfr | vfr gene deletion on pEX18Tc; Tcr | 58 |
| pETMAL-osaR | pETMALc-H vector carrying the malE-OsaR fusion; Kanr | 20 |
| pUCP19 | Multicopy E. coli-P. aeruginosa shuttle vector; Apr/Cbr | 59 |
| pUCP19-osaR | pUCP19 carrying the intact osaR gene; Apr/Cbr | 20 |
| pKF912 | pUCP19 carrying vfr as a 1.2-kb XhoI fragment; Apr/Cbr | 42 |
| pDN19lacΩ | Broad-host-range plasmid containing a promoterless lacZ gene; Tcr | 60 |
| PfleQ-lacZ | pDN19lacΩ carrying the promoter region of fleQ in the EcoRI/BamHI sites; Tcr | This study |
| PfleQm1-lacZ | PfleQ-lacZ with the first A-T pair of the OsaR binding site mutated; Tcr | This study |
| PfleQm2-lacZ | PfleQ-lacZ with the second A-T pair of the OsaR binding site mutated; Tcr | This study |
| PfleQm3-lacZ | PfleQ-lacZ with the two A-T pairs of the OsaR binding site mutated; Tcr | This study |