Table 1.
Bacterial strains and plasmids used in this study.
| Strain or plasmid | Description | Source |
|---|---|---|
| E. coli strains | ||
| DH5α | F- ϕ 80dlacZΔM15 endA1 recA1 hsdR17(rK- mK+) supE44 thi-1 relA1 Δ(lacZYA-argF)U169 gyrA96 deoR | TransGen |
| S17-1 | RP4-2 Tc::Mu Km::Tn7 Tpr Smr Pro Res- Mod+ | Dr. Ramphal |
| HB101 | Source for wild type lacP1 promoter sequence | TransGen |
| P. aeruginosa strains | ||
| PA14 | Wild type P. aeruginosa strain | Liberati et al., 2006 |
| PAK | Wild type P. aeruginosa strain | David Bradley |
| exsA::Tn | PA14 with exsA disrupted by insertion of Tn | Liberati et al., 2006 |
| PA4336::Tn | PA14 with PA4336 inserted with Tn | Liberati et al., 2006 |
| PA4916::Tn | PA14 with PA4916 inserted with Tn | Liberati et al., 2006 |
| PA0020::Tn | PA14 with PA0020 inserted with Tn | Liberati et al., 2006 |
| PA4753::Tn | PA14 with PA4753 inserted with Tn | Liberati et al., 2006 |
| exsA::Ω | PAK with exsA disrupted by insertion of cassette Ω; Spr, Smr | Li et al., 2013 |
| ΔnadD2 | PAK with nadD2 deleted | This study |
| ΔnadD2-nrtR | PAK with nadD2-nrtR operon deleted | This study |
| ΔPA4916 | PAK with PA4916 deleted | This study |
| ΔPA4916/att7::PA4916 | ΔPA4916 with PA4916 inserted into the chromosome with mini-Tn7 insertion | This study |
| Δvfr | PAK with vfr gene deleted | This study |
| ΔnrtRΔvfr | PAK with both nrtR and vfr gene deleted | This study |
| Plasmids | ||
| pDN19 | Shuttle vector between E. coli and P. aeruginosa; Tcr | Li et al., 2013 |
| pMMB67EH | Shuttle vector between E. coli and P. aeruginosa; Ampr | Li et al., 2016 |
| pE1553 | Promoterless pUCP20; Ampr | Li et al., 2016 |
| pE1553a | cyaA-flag with own promoter in pE1553; Ampr | This study |
| pE1553b | cyaB-flag with own promoter in pE1553; Ampr | This study |
| pE1553-cpdA | cpdA-flag with own promoter in pE1553; Ampr | This study |
| pMMB67-cyaA | cyaA-flag cloned into pMMB67EH driven by tac promoter; Ampr | This study |
| pMMB67-cyaB | cyaB-flag cloned into pMMB67EH driven by tac promoter; Ampr | This study |
| pMMB67-nadD2 | nadD2 gene of PAK on pMMB67EH driven by tac promoter; Ampr | This study |
| pMMB67-4918-20 | PA4918-20 gene of PAK on pMMB67EH driven by tac promoter; Ampr | This study |
| pMMB67EH-cyaB217-463-His | CyaB217-463-His cloned into pMMB67EH driven by tac promoter; Ampr | This study |
| pMMB67EH-cyaA-His | CyaA-His cloned into pMMB67EH driven by tac promoter; Ampr | This study |
| pUCP24 | Shuttle vector between E. coli and P. aeruginosa; Gmr | Shi et al., 2015 |
| pUCP24-nadD2-Flag | nadD2-Flag cloned into pUCP24; Gmr | This study |
| exsA | exsA gene of PAK on pDN19 driven by lac promoter; Tcr | Li et al., 2013 |
| lacP1 | LacP1 promoter of E. coli fused to promoterless lacZ on pDN19lacZΩ; Spr, Smr, Tcr | This study |
| pUC18T-mini-Tn7T-Gm | mini-Tn7 base vector insertion into chromosome attTn7 site, Gmr | Li et al., 2013 |
| pUC18T-mini-Tn7T-PexsC-exsCEBA-Flag-ExsD | exsCEBAD gene with exsA-Flag tagged on pUC18T-mini-Tn7T driven by exsC promoter, Gmr | This study |
| pTNS3 | Helper plasmid encoding Tn7 site-specific transposition pathway; Ampr | Deng et al., 2017 |
| pEX18Tc | Gene knockout vector; Tcr | Deng et al., 2017 |
| pZF01 | PA4916 gene deletion on pEX18Tc; Tcr | This study |
| pEX18Tc-ΔnadD2 | nadD2 gene deletion on pEX18Tc; Tcr | This study |
| pEX18Tc-ΔnadD2-nrtR | nadD2-nrtR operon deletion on pEX18Tc; Tcr | This study |
| pEX18Tc-Δvfr | vfr gene deletion on pEX18Tc; Tcr | This study |
| pEX18Tc-ΔcyaA | cyaA gene deletion on pEX18Tc; Tcr | This study |
| pEX18Tc-ΔcyaB | cyaB gene deletion on pEX18Tc; Tcr | This study |
| pZF02 | PA4916 gene on pUC18T-mini-Tn7T-Tc; Tcr | This study |
| pET16b | Expression vector, Kanr | Novagen |
| pET16b-nadD2 | nadD2 gene of PAK cloned into pET16b | This study |
| pET28a | Expression vector, Kanr | Novagen |
| pET28a-cyaB217-463 | cyaB gene encoding amino acid positions 217–463 cloned into pET28a | This study |