TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant characteristics | Reference or source |
|---|---|---|
| P. aeruginosa | ||
| PAO1 | Reference strain completely sequenced | 40 |
| PAΔD | PAO1 ΔampD::lox | This work |
| PAΔDh2 | PAO1 ΔampDh2::lox | This work |
| PAΔDh3 | PAO1 ΔampDh3::lox | This work |
| PAΔDDh2 | PAO1 ΔampD::lox ΔampDh2::lox | This work |
| PAΔDDh3 | PAO1 ΔampD::lox ΔampDh3::lox | This work |
| PAΔDh2Dh3 | PAO1 ΔampDh2::lox ΔampDh3::lox | This work |
| PAΔDDh2Dh3 | PAO1 ΔampD::lox ΔampDh2::lox ΔampDh3::lox | This work |
| E. coli | ||
| XL1-Blue | F′::Tn10 proA+B+lacIq Δ(lacZ)M15 recA1 endA1 gyrA96 (Nalr) thi hsdR17(rK− mK−) mcrB1 | 3 |
| S17-1 | RecA pro (RP4-2Tet::Mu Kan::Tn7) | 37 |
| Plasmids | ||
| pGEMT | PCR cloning vector | Promega |
| pUCP24 | Gmr; pUC18-based Escherichia-Pseudomonas shuttle vector | 42 |
| pUCPAD | Gmr; pUCP24 containing PAO1 ampD gene | 18 |
| pUCPADE | Gmr; pUCP24 containing complete PAO1 ampDE operon | 18 |
| pUCPADh2 | Gmr; pUCP24 containing PAO1 ampDh2 gene | This work |
| pUCPADh3 | Gmr; pUCP24 containing PAO1 ampDh3 gene | This work |
| pEX100Tlink | AprsacB; pUC19-based gene replacement vector with MCSa | 36 |
| pUCGmlox | Apr Gmr; pUC18-based vector containing lox-flanked aacC1 gene | 36 |
| pCM157 | Tcr; cre expression vector | 31 |
| pEXADGm | pEX100Tlink containing 5′ and 3′ flanking sequence of ampD::Gmlox | This work |
| pEXADh2Gm | pEX100Tlink containing 5′ and 3′ flanking sequence of ampDh2::Gmlox | This work |
| PEXADh3Gm | pEX100Tlink containing 5′ and 3′ flanking sequence of ampDh3::Gmlox | This work |
a
MCS, multiple cloning site.