TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | F− φ80dlac ΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 (rK− mK−) supE44 λ− thi-1 gyrA96 relA1 | Bethesda Research Laboratory |
| P. aeruginosa | ||
| PAO1 | Wild type | D. Haas |
| PAO1214 | aotJ::Tn5-751 insertion mutant of PAO1 | 22 |
| PAO501 | argR; gentamicin resistant | 22 |
| PAO4447 | leu; kanamycin sensitive | Y. Itoh |
| PAO4449 | aotO::aph mutant of PAO4447 | This study |
| PAO4450 | aot::aph mutant of PAO4447; kanamycin resistant | This study |
| Plasmids | ||
| pUC119 | bla lacZ′ cloning vector | 27 |
| pUC19 | bla lacZ′ cloning vector | 31 |
| pQF52 | bla lacZ translational fusion vector | 23 |
| pST500 | aotJ::lacZ translational fusion of pQF52 | This study |
| pST26 | 360-bp PCR fragment of the aot promoter region cloned into SmaI site of pUC19 | This study |
| pMO011644 | Cosmid clone carrying the aot-argR and aru operons of P. aeruginosa | 13 |
| pYJ61 | 10-kb insert in pMO011644 subcloned into HincII site of pUC119 | This study |
| pYJ62 | Similar to pYJ61, but with the opposite orientation | This study |
| pTnL1 | Clone of PAO1214 carrying the aotJ regulatory region and the aph gene of Tn5-751 | This study |