TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype and/or relevant characteristics | Reference or source |
|---|---|---|
| P. aeruginosa strains | ||
| PAO1 (ATCC 15692) | Prototroph | American Type Culture Collection |
| PAO1 ΔrhdA | rhdA null mutant | This study |
| PAO1-Nal | Nalr | This study |
| PAO1-Rif | Rifr | This study |
| PAO1-Rif ΔrhdA | PAO1 ΔrhdA, Rifr | This study |
| Plasmids | ||
| pEX18Tc | PMB1 replicon, oriT sacB lacZα, Mob+ Tcr, allelic exchange vector | 23 |
| pFLP2 | AprsacB; broad-host-range recombination system | 23 |
| pPS858 | Apr; source of Gmr-GFP cassette | 23 |
| pEXΔrhdA | pEX18Tc derivative carrying the Gmr-GFP cassette between rhdA flanking regions | This study |
| pUCP18 | E. coli-Pseudomonas shuttle vector derived from pUC18/19; ColE1, pRO1600 replicon, lacZ bla, Apr Cbr | 34 |
| pUCPrhdA | 1,002-bp PCR product, encompassing the rhdA coding sequence and its putative promoter, ligated to the BamHI-HindIII sites of pUCP18 | This study |
| pUCP4955-rhdA | 2,645-bp PCR product encompassing both PA4955 and rhdA coding sequences with their putative promoters, ligated to the BamHI-HindIII sites of pUCP18 | This study |
| pMP220 | Broad-host-range, low-copy-number promoter-probe vector; IncP replicon, lacZ Tcr Tra− | 38 |
| pMP4955 | 322-bp PCR product encompassing the putative PA4955 promoter, ligated to the BglII-PstI sites of pMP220 | This study |
| pMPrhdA | 305-bp PCR product encompassing the putative rhdA promoter, ligated to the BglII-PstI sites of pMP220 | This study |
| pMP4955-rhdA | 1,948-bp PCR product encompassing both PA4955 and rhdA putative promoters with the PA4955 intervening ORF, ligated to the BglII-PstI sites of pMP220 | This study |
| pMP220R | Derivative of pMP220 with polylinker in inverted orientation | This study |
| pMPR4955 | Same insert of pMP4955 cloned in pMP220R | This study |
| pMPRrhdA | Same insert of pMPrhdA cloned in pMP220R | This study |
| pMPR4955-rhdA | Same insert of pMP4955-rhdA cloned in pMP220R | This study |