Table 1.
Bacterial strains and plasmids used in this study.
| Strain or plasmid | Genotype and/or relevant characteristics | Reference or source |
|---|---|---|
| STRAINS | ||
| P. aeruginosa | ||
| PAO1 | ATCC15692 (wild type, prototroph) | American type culture collection |
| ΔhasR | PAO1ΔhasR | This work |
| ΔphuR | PAO1ΔphuR | This work |
| ΔhasRΔphuR | PAO1ΔhasRΔphuR | Minandri et al., 2016 |
| ΔpvdA | PAO1ΔpvdA | Imperi et al., 2008 |
| ΔpchD | PAO1ΔpchD | Frangipani et al., 2014 |
| ΔpvdAΔpchD | PAO1ΔpvdAΔpchD | Visca et al., 2013 |
| Δcio | PAO1 containing a 2400-bp deletion in the cioAB locus | This work |
| Δcox | PAO1 containing a 4109-bp deletion in the coxBA-PA0107-coIII locus | This work |
| Δcyo | PAO1 containing a 4830-bp deletion in the cyoABCDE operon | This work |
| Δcco | PAO1 containing a 6445-bp deletion in the two adjacent ccoNOQP1 and ccoNOQP2 operons | This work |
| ΔcyoΔcio | PAO1 mutated in both cyo and cio | This work |
| ΔcyoΔcco | PAO1 mutated in both cyo and cco-1,2 | This work |
| ΔcyoΔcioΔcox | PAO1 mutated in cyo, cio and cox | This work |
| ΔcyoΔccoΔcox | PAO1 mutated in cyo, cco and cox | This work |
| E. coli | ||
| DH5αF′ | recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1 Δ(lacZYA-argF) U169 [Φ80dlacZΔM15] NalR | Sambrook et al., 1989 |
| S17-1 λpir | recA, thi, pro, hsdR-M+RP4: 2-Tc:Mu: Km Tn7 λpir, TpR SmR | Simon et al., 1983 |
| PLASMIDS | ||
| pDM4 | Suicide vector; sacBR, oriR6K, CmR | Milton et al., 1996 |
| pME7541 | Suicide construct used for deletion of the cioAB operon; TcR | Frangipani et al., 2008 |
| pME9302 | Suicide construct used for deletion of the coxB-coIII cluster; TcR | Frangipani et al., 2008 |
| pME9303 | Suicide construct used for deletion of the cyoABCDE operon; TcR | Frangipani et al., 2008 |
| pME9308 | Suicide construct used for deletion of the two adjacent ccoNOQP operons; TcR | Frangipani et al., 2008 |
| pUCP18 | E. coli-Pseudomonas shuttle vector derived from pUC18; ColE1, pRO1600, ApR, CbR | Schweizer, 1991 |
| pUCPhasR | pUCP18 derivative carrying the coding sequence of hasR with its own promoter | This study |
| pUCPphuR | pUCP18 derivative carrying the coding sequence of phuR with its own promoter | This study |
| pUCPphuRhasR | pUCP18 derivative carrying the coding sequence of phuR and hasR with their own promoters | Minandri et al., 2016 |
| pDM4ΔhasR | pDM4 derivative carrying the flanking regions of the hasR coding sequence | Minandri et al., 2016 |
| pDM4ΔphuR | pDM4 derivative carrying the flanking regions of the phuR coding sequence | Minandri et al., 2016 |
| Oligonucleotides | Sequence 5′-3′ | Restriction site |
| hasR compl FW | CGGGGTACCGGCGGGAGTGACGCTGC | KpnI |
| hasR compl RV | GAAGATCTCCTTCACTGGGCAAAACGG | BglII |
| phuR compl FW | CCGGAATTCGAAAGGCTGGGAGTGCTG | EcoRI |
| phuR compl RV | CGGGGTACCACCTGTGGCATGGAAAGC | KpnI |
TcR, tetracycline resistant; ApR, ampicillin resistant; CmR, chloramphenicol resistant; CbR, carbenicillin resistant; restriction sites in the oligonucleotides are underlined.