TABLE 1.
Strains and plasmids used
| Strain or plasmid | Relevant genotype or phenotype | Reference or source |
|---|---|---|
| E. coli DH5α | F′/endA1 hsdR17 supE44 thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169 deoR [φ80 dlacZΔM15 recA1] | 40 |
| P. aeruginosa | ||
| PAO1 | Wild-type, elastolytic, prototroph | 12 |
| PAO-R1 | ΔlasR::Tcr, nonelastolytic derivative of strain PAO1 | 8 |
| PR1-E4 | ΔlasR::Tcr, casein incubation-induced elastolytic mutant of strain PAO-R1 | 36 |
| PDO100 | ΔrhlI::Tn501-2 derivative of PAO1 | 3 |
| Plasmids | ||
| pBluescript II SK+ | General-purpose cloning vector, ori (ColE1), Apr | Stratagene |
| pUC18, pUC19 | General-purpose cloning vectors, ori (ColE1) | Gibco |
| pUCP18, pUCP19, pUCP24 | E. coli-P. aeruginosa shuttle vectors | 30 |
| pRL1500 | pUCP19 containing rhlA on a 1.5-kb BglI-NcoI fragment from strain PG201 | 18 |
| pPAO-2 | pUC19 containing a 280-bp Sau3A fragment of pilA from strain PAO1 | 29 |
| pJPP41 | pBluescript II SK+ containing rhlI on a 950-bp BglII-SphI fragment from strain PAO1 | 25 |
| pRB1801 | pUC18 containing a 1.75-kb EcoRI-SmaI fragment of lasB from strain PAO1 | 2 |
| GB24 | P. aeruginosa gene bank: partial Sau3A digest of strain PAO1 DNA inserted into the BamHI site of pUCP24 | U. Ochsner |
| pVD99.0 | pUCP24 containing a 2.5-kb Sau3A genome fragment from strain PAO1 | This study |
| pVD99.1 | pUCP24 containing a 1.98-kb EcoRI fragment from pVD99.0 | This study |
| pVD99.3 | pUCP24 containing dksA on a 960-bp SmaI fragment from pVD99.0 | This study |
| pVD99.4 | pUCP24 containing a 523-bp EcoRI fragment from pVD99.0 | This study |
| pVD99.5 | pUCP24 containing a 520-bp SalI fragment from pVD99.0 | This study |
| pECP60 | pSW205 containing a rhlA′-lacZ translational fusion | 26 |
| pDECP60 | pECP60 containing dksA on a 960-bp SmaI fragment from pVD99.3 | This study |
| pLPR1 | pLP170 containing a rhlI′-lacZ transcriptional fusion | 36 |
| pVD | pLPRI containing dksA on a 960-SmaI fragment from pVD99.3 | This study |
| pPBL25 | pUCP18 containing lasB′-lacZ on a 3-kb Eco0109 fragment from pKDT37 (22) | This study |
| pPBLD26 | pPBL25 containing dksA on a 960-bp SmaI fragment from pVD99.3 | This study |