TABLE 1.
Strains and plasmids used in this study
| Name | Genotype or description | Reference |
|---|---|---|
| Strains | ||
| PA14 | P. aeruginosa burn wound isolate | 67 |
| TFM15 | PA14 ΔndvB; unmarked deletion of ndvB (codons 273–727 deleted) | 18 |
| TFM425 | PA14 ΔrpoS; unmarked deletion of rpoS (codons 15–272 deleted) | This study |
| TFM423 | PA14 attTn7::mini-Tn7T-Gm-lacZ; promoterless lacZ; Gmr | This study |
| TFM427 | PA14 attTn7::mini-Tn7T-Gm-PndvB-lacZ; PndvB-lacZ fusion; Gmr | This study |
| TFM456 | PA14 attTn7::mini-Tn7T-Gm-PndvB-mut-lacZ; PndvB-lacZ fusion with mutated RpoS binding site; Gmr | This study |
| DH5α | E. coli strain; supE44 ΔlacU169(ϕ80 lacZΔM15) recA1 hsdR17 thi-1 relA1 | 81 |
| S17-1 | E. coli strain; recA pro hsdR RP4-2-Tc::Mu-Km::Tn7 | 82 |
| YPH500 | Saccharomyces cerevisiae uracil auxotroph; MATα ura3-52 lys2-801amber ade2-101ochre trp1-Δ63 his3-Δ200 leu2-Δ1 | 83 |
| Plasmids | ||
| pMQ80 | araC-PBAD-gfpmut3 reporter on E. coli-S. cerevisiae-Pseudomonas shuttle vector; Gmr | 33 |
| pSC01 | PndvB-gfp stable gfp reporter; pMQ80 with ndvB promoter upstream of gfpmut3; Gmr | This study |
| pAH04 | araC-PBAD-gfp (ASV) reporter; pMQ80 with gfp protease tag from pJBA113; Gmr | This study |
| pAH05 | PndvB-gfp (LAA) reporter; pSC01 with gfp protease tag from pJBA110; Gmr | This study |
| pAH06 | PndvB-gfp (LVA) reporter; pSC01 with gfp protease tag from pJBA111; Gmr | This study |
| pAH07 | PndvB-gfp (AAV) reporter; pSC01 with gfp protease tag from pJBA112; Gmr | This study |
| pAH08 | PndvB-gfp (ASV) reporter; pSC01 with gfp protease tag from pJBA113; Gmr | This study |
| pJBA110 | pUC18Not-PA1/04/03-RBSII-gfp (LAA)-T0-T1; source of LAA protease tag; Apr | 34 |
| pJBA111 | pUC18Not-PA1/04/03-RBSII-gfp (LVA)-T0-T1; source of LVA protease tag; Apr | 34 |
| pJBA112 | pUC18Not-PA1/04/03-RBSII-gfp (AAV)-T0-T1; source of AAV protease tag; Apr | 34 |
| pJBA113 | pUC18Not-PA1/04/03-RBSII-gfp (ASV)-T0-T1; source of ASV protease tag; Apr | 34 |
| pEX18Gm | ||
| pEX18Gm::ΔrpoS | pEX18Gm derivative for unmarked deletion of rpoS; Gmr | This study |
| pUC18-mini-Tn7T-Gm-lacZ | Mini-Tn7T transcriptional lacZ fusion vector; Apr Gmr | 68 |
| pUC18-mini-Tn7T-Gm-PndvB-lacZ | pUC18-mini-Tn7T-Gm-lacZ carrying the sequence from −112 to −1 relative to the translational start site of ndvB; Apr Gmr | This study |
| pUC18-mini-Tn7T-Gm-PndvB-mut-lacZ | Same as pUC18-mini-Tn7T-Gm-PndvB-lacZ except that the sequence CTAGACT was mutated to AGAGACT; Apr Gmr | This study |
| pTNS2 | Transposase helper plasmid for mini-Tn7 integration; Apr | 68 |
| pRK2013 | Helper plasmid for triparental matings; Kmr | 72 |
| pUC18 | High-copy-number E. coli cloning/sequencing plasmid; Apr | 84 |
| pUCP19 | Broad-host-range cloning and expression vector; Cbr | 85 |
| pUCP19::rpoS+ | pUCP19 with rpoS under the control of its native promoter and terminator; Cbr | This study |