TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| DH5α | endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 (argF-lacZYA)U169 φ80dlacZ | 42 |
| S17-1 | hsdR pro recA; contains RP4-2-Tc::Mu integrated into the chromosome | 47 |
| SNO301-1 | ampD1 ampG | 45 |
| Pseudomonas aeruginosa | ||
| PAO1 | Reference strain; genome completely sequenced | 48 |
| PAO1ΔDDh2Dh3 | PAO1 deleted of the three ampD homologues | 21 |
| PAK | Reference strain | David Bradley |
| PAO1ΔG | PAO1 deleted of ampG (PA4393) | This study |
| PAO1ΔGh1 | PAO1 deleted of ampGh1 (PA4218) | This study |
| PAO1ΔGGh1 | PAO1 deleted of both ampG (PA4393) and ampGh1 (PA4218) | This study |
| PAKΔG | PAK deleted of ampG (PA4393) | This study |
| PAKΔGh1 | PAK deleted of ampGh1 (PA4218) | This study |
| PAKΔGGh1 | PAK deleted of both ampG (PA4393) and ampGh1 (PA4218) | This study |
| PAO1ΔDDh2Dh3G | PAO1ΔDDh2Dh3 deleted of ampG (PA4393) | This study |
| PAO1ΔDDh2Dh3Gh1 | PAO1ΔDDh2Dh3 deleted of ampGh1 (PA4218) | This study |
| Plasmids | ||
| pGEMT | PCR cloning vector | Promega |
| pEC1C | ampC ampR from E. cloacae; Cmr | 16 |
| pGKS273-5 | ampG region from E. coli JRG582; Kmr | 28 |
| pUCP24 | pUC18-derived broad-host-range vector; Gmr | 52 |
| pUCP26 | pUC18-derived broad-host-range vector; Tcr | 52 |
| pEX18Tc | Counter selectable plasmid carrying sacB marker; oriT; Tcr | 14 |
| pEXAG | pEX18Tc carrying ampG deletion construct; Tcr | This study |
| pEXAGh1 | pEX18Tc carrying ampGh1 deletion construct; Tcr | This study |
| pZY0901 | ampG gene of PAO1 cloned into pUCP24; Gmr | This study |
| pZY0908 | ampGh1 gene of PAO1 cloned into pUCP24; Gmr | This study |
a
Antibiotic resistance markers: Cm, chloramphenicol; Gm, gentamicin; Km, kanamycin; Tc, tetracycline.