TABLE 1.
Bacterial strains and plasmids used in this study
| Strains or plasmids | Relevant genotype or phenotype | Source or reference |
|---|---|---|
| P. aeruginosa strains | ||
| PAO1 | Wild-type strain | Lab collection |
| ΔfleR | PAO1 with the deletion of the fleR gene | This study |
| ΔfleR(EV) | ΔfleR containing the empty plasmids pBBR1-MCS5 or pUCP18 | This study |
| ΔfleR(fleR) | ΔfleR containing the expression constructs pBBR1-MCS5-fleR or pUCP18-fleR | This study |
| ΔfleR(sadC) | ΔfleR containing the expression constructs pBBR1-MCS5-sadC or pUCP18-sadC | This study |
| ΔfleR(bifA) | ΔfleR containing the expression constructs pBBR1-MCS5-bifA or pUCP18-bifA | This study |
| ΔfleR(14945) | ΔfleR containing the expression constructs pBBR1-MCS5-W909_14945 or pUCP18-W909_14945 | This study |
| ΔfleR(14950) | ΔfleR containing the expression constructs pBBR1-MCS5-W909_14950 or pUCP18-W909_14950 | This study |
| ΔfleQ | PAO1 with the deletion of the fleQ gene | This study |
| ΔfleQ(EV) | ΔfleQ containing the empty plasmids pBBR1-MCS5 or pUCP18 | This study |
| ΔfleQ(sadC) | ΔfleQ containing the expression construct pBBR1-MCS5-sadC | This study |
| ΔfleRΔfleQ | PAO1 with the deletion of the fleR and fleQ genes | This study |
| ΔfleRΔfleQ(EV) | ΔfleRΔfleQ containing the empty plasmids pBBR1-MCS5 or pUCP18 | This study |
| ΔfleRΔfleQ(sadC) | ΔfleRΔfleQ containing the expression constructs pBBR1-MCS5-sadC or pUCP18-sadC | This study |
| Δtse1 | PAO1 with the deletion of the tse1 gene | This study |
| Δtse1(EV) | Δtse1 containing the empty plasmids pBBR1-MCS5 or pUCP18 | This study |
| Δtse1(tse1) | Δtse1 containing the expression construct pUCP18-tse1 | This study |
| ΔfleRΔtse1 | PAO1 with the deletion of the fleR and tse1 genes | This study |
| ΔfleRΔtse1(tse1) | ΔfleRΔtse1 containing the expression construct pUCP18-tse1 | This study |
| ΔsadC | PAO1 with the deletion of the sadC gene | This study |
| ΔfleRΔsadC | PAO1 with the deletion of the fleR and sadC genes | This study |
| ΔfleRΔflgZ | PAO1 with the deletion of the fleR and flgZ genes | This study |
| ΔfleRΔflgZ(sadC) | ΔfleRΔflgZ containing the expression constructs pBBR1-MCS5-sadC or pUCP18-sadC | This study |
| PAO1(amrZ) | PAO1 containing the expression constructs pBBR1-MCS5-amrZ or pUCP18-amrZ | This study |
| ΔfleR(amrZ) | ΔfleR containing the expression constructs pBBR1-MCS5-amrZ or pUCP18-amrZ | This study |
| ΔfleRΔfleQ(amrZ) | ΔfleRΔfleQ containing the expression constructs pBBR1-MCS5-amrZ or pUCP18-amrZ | This study |
| ΔretS | PAO1 with the deletion of the retS gene | This study |
| ΔretS(EV) | ΔretS containing the empty plasmids pBBR1-MCS5 or pUCP18 | This study |
| ΔfleRΔretS | PAO1 with the deletion of the fleR and retS genes | This study |
| ΔfleRΔretS(EV) | ΔfleRΔretS containing the empty plasmids pBBR1-MCS5 or pUCP18 | This study |
| ΔfleRΔretS(retS) | ΔfleRΔretS containing the expression constructs pBBR1-MCS5-retS or pUCP18-retS | This study |
| E. coli strains | ||
| DH5α | spuE44 ΔlacU169(φ80lacZΔM15) hsdR17λpir recA1 endA1 gyrA96 thi-1 relA1 | Lab collection |
| pRK2013 | Tra+, Mob-, ColE1-replicon, Kanr, Sper | Lab collection |
| Plasmids | ||
| pBBR1-MCS5 | Broad-host-range cloning vector; Gmr | Lab collection |
| pUCP18 | E. coli-P. aeruginosa shuttle expression vector with Plac, Ampr, Carr | Lab collection |
| pBBR1-MCS5-fleR | pBBR1-MCS5 containing fleR under the control of Plac | This study |
| pUCP18-fleR | pUCP18 containing fleR under the control of Plac | This study |
| pBBR1-MCS5-sadC | pBBR1-MCS5 containing sadC under the control of Plac | This study |
| pUCP18-sadC | pUCP18 containing sadC under the control of Plac | This study |
| pBBR1-MCS5-bifA | pBBR1-MCS5 containing bifA under the control of Plac | This study |
| pUCP18-bifA | pUCP18 containing bifA under the control of Plac | This study |
| pUCP18-tse1 | pUCP18 containing tse1 under the control of Plac | This study |
| pBBR1-MCS5-W909_14945 | pBBR1-MCS5 containing W909_14945 under the control of Plac | This study |
| pUCP18-W909_14945 | pUCP18 containing W909_14945 under the control of Plac | This study |
| pBBR1-MCS5-W909_14950 | pBBR1-MCS5 containing W909_14950 under the control of Plac | This study |
| pUCP18-W909_14950 | pUCP18 containing W909_14950 under the control of Plac | This study |
| pBBR1-MCS5-retS | pBBR1-MCS5 containing retS under the control of Plac | This study |
| pUCP18-retS | pUCP18 containing retS under the control of Plac | This study |
| pBBR1-MCS5-amrZ | pBBR1-MCS5 containing amrZ under the control of Plac | This study |
| pUCP18-amrZ | pUCP18 containing amrZ under the control of Plac | This study |
| pK18mobsacB | Broad-host-range sucrose counter-selection allelic exchange vector, sacB, Gmr | Lab collection |
| pK18-fleR | pK18 containing fleR flanking regions for generation of fleR in-frame deletion | This study |
| pK18-tse1 | pK18 containing tse1 flanking regions for generation of tse1 in-frame deletion | This study |
| pK18-retS | pK18 containing retS flanking regions for generation of retS in-frame deletion | This study |
| pK18-fleQ | pK18 containing fleQ flanking regions for generation of fleQ in-frame deletion | This study |
| pK18-flgZ | pK18 containing flgZ flanking regions for generation of flgZ in-frame deletion | This study |
| pK18-sadC | pK18 containing sadC flanking regions for generation of sadC in-frame deletion | This study |
| pK18-tse1-his | pK18 containing tse1. Six repeated CAC was added at the C-terminal. | This study |
| pK18-hcp1-his | pK18 containing hcp1. Six repeated CAC was added at the C-terminal. | This study |