TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype and/or relevant characteristics | Reference or source |
|---|---|---|
| Strains | ||
| P. aeruginosa strains | ||
| PAO1 | ATCC 15692 (wild type, prototroph) | American Type Culture Collection |
| PAO1 PpvdA′-′mCherry | Wild type with PpvdA::mCherry | This work |
| pvdA mutant | PAO1 ΔpvdA | 45 |
| pchD mutant | PAO1 ΔpchD | 82 |
| pvdA pchD mutant | PAO1 ΔpvdA ΔpchD | 83 |
| PAO1 pvdA pchD PpvdA′-′mCherry | PAO1 ΔpvdA ΔpchD PpvdA::mCherry | This work |
| fpvR mutant | PAO1 ΔfpvR | This work |
| pvdA fpvR mutant | PAO1 ΔpvdA ΔfpvR | This work |
| pvdS mutant | PAO1 ΔpvdS Gmr | 31 |
| hasR phuR mutant | PAO1 ΔhasR ΔphuR | This work |
| feoB mutant | PAO1 ΔfeoB | This work |
| tonB1 mutant | PAO1 ΔtonB1 | This work |
| tonB1 feoB mutant | PAO1 ΔtonB1 ΔfeoB | This work |
| toxA mutant | PAO1 phoA wp03q2B07 (mutant 40695) | Transposon mutant collection of University of Washington |
| Escherichia coli strains | ||
| DH5α | F′ recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1 Δ(lacZYA-argF)U169 ϕ80dlacZΔM15 Nalr | 35 |
| S17-1 λpir | recA thi pro hsdR(M+ RP4)::2-Tc::Mu::Km Tn7 λpir Tpr Smr | 84 |
| Plasmids | ||
| pEX18Tc | Allelic exchange vector pMB1 replicon; oriT+ sacB+ lacZα mob+ Tcr | 39 |
| pDM4 | Suicide vector; sacBR oriR6K Cmr | 40 |
| pmCherry-1 | Vector used as template for amplification of the mCherry coding sequence | Clontech |
| Mini-CTX1 | Promoter-probe vector; Ω-FRT-attP-MCS ori int oriT Tcr | 42 |
| pFLP2 | FLP recombinase-expressing plasmid; Apr | 39 |
| Mini-CTX1 mCherry | Tcr | This study |
| Mini-CTX PpvdA′-′mCherry | Plasmid to insert a PpvdA-mCherry fusion into the chromosome of P. aeruginosa; Tcr | This study |
| pDM4ΔfpvR | pDM4 derivative carrying the flanking regions of the fpvR coding sequence | This study |
| pDM4ΔhasR | pDM4 derivative carrying the flanking regions of the hasR coding sequence | This study |
| pDM4ΔphuR | pDM4 derivative carrying the flanking regions of the phuR coding sequence | This study |
| pEXΔfeoB | pEX18Tc derivative carrying a Gmr-GFP cassette between the feoB flanking regions | This study |
| pDM4ΔtonB | pDM4 derivative carrying the flanking regions of the tonB1 coding sequence | This study |
| pMP220::PfeoA | pMP220 derivative carrying a PfeoA::lacZ transcriptional fusion; Tcr | 31 |
| pPZ-toxA | pPZ20 derivative carrying a PtoxA::lacZ translational fusion; Cbr | 26 |
| pPZ-prpL | pPZTC derivative carrying a PprpL::lacZ transcriptional fusion; Cbr | 26 |
| pUCP18 | E. coli-Pseudomonas shuttle vector derived from pUC18; ColE1 pRO1600 Apr Cbr | 41 |
| pUCPpvdA | pUCP18 derivative carrying the coding sequence of pvdA with its own promoter | This study |
| pUCPpchD | pUCP18 derivative carrying the coding sequence of pchD with its own promoter | This study |
| pUCPtonB1 | pUCP18 derivative carrying the coding sequence of tonB1 with its own promoter | This study |
| pUCPphuRhasR | pUCP18 derivative carrying the coding sequences of phuR and hasR with their own promoters | This study |