TABLE 4.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Description or sequence (5′→3′) | Source, reference,purpose, or function |
|---|---|---|
| P. aeruginosa strains | ||
| PAK | Wild-type strain of Pseudomonas aeruginosa | David Bradley |
| ΔKH-S1 | PAK with pnp (KH and S1) deletion | 38 |
| ΔKH-S1/Tn7T-pnp | PAK ΔKH-S1 with pnp inserted on the chromosome with a mini-Tn7T insertion | 38 |
| PAK ΔmexXY | PAK deleted of mexXY | This study |
| PAK ΔarmZ | PAK deleted of armZ | This study |
| ΔKH-S1 ΔmexXY | PAK ΔKH-S1 deleted of mexXY | This study |
| ΔKH-S1 ΔarmZ | PAK ΔKH-S1 deleted of armZ | This study |
| PAK ΔmexZ | PAK deleted of mexZ | This study |
| Plasmids | ||
| pEX18Tc | Gene replacement vector; Tcr oriT+ sacB+ | 58 |
| pMMB67EH | Expression vector with a tac promoter; Apr | 59 |
| pMMB67EH-mexZ | MexZ on pMMB67EH | This study |
| pUCP20(no promoter) | Escherichia-Pseudomonas shuttle vector; no promoter; Ampr | This study |
| pUCP20(no promoter)-PmexZ-LacZ | mexZ promoter and lacZ gene on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed-PA5471.1-armZ-His | 6×His-tagged armZ driven by the PBAD promoter with 275 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed(pET28a)-armZ-His | 6×His-tagged armZ driven by the PBAD promoter with the ribosome binding site from pET28a on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed-armZ-His | 6×His-tagged armZ driven by the PBAD promoter with 212 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed(−86)-armZ-His | 6×His-tagged armZ driven by the PBAD promoter with 86 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed(−36)-armZ-His | 6×His-tagged armZ driven by the PBAD promoter with 36 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed(−15)-armZ-His | 6×His-tagged armZ driven by the PBAD promoter with 15 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed(−36)-GFP | GFP driven by the PBAD promoter with 36 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| pUCP20(no promoter)-pRkaraRed(−15)-GFP | GFP driven by the PBAD promoter with 15 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
| Primers | ||
| mexA-RT-S | AGGTGCGTCCCCAGGTG | RT-PCR |
| mexA-RT-AS | CCTCGTAGGTGGCGGG | RT-PCR |
| mexB-RT-S | AACGTGCAGATTTCCTCCG | RT-PCR |
| mexB-RT-AS | GATGTTCTCGAATTGCTCCG | RT-PCR |
| mexX-RT-S | TGCGAAGAAGCAGCGGA | RT-PCR |
| mexX-RT-AS | CAGGCGACGGGTGACG | RT-PCR |
| mexY-RT-S | CTACAACATCCCCTATGACACCT | RT-PCR |
| mexY-RT-AS | CATCACGGCGAACACCAG | RT-PCR |
| mexZ-RT-S | ACTACAAGAACAAGATCGAGG | RT-PCR |
| mexZ-RT-AS | TGGCGTTTTCGTCGGGTA | RT-PCR |
| armZ-RT-S | GCAAACCACCGCCGAA | RT-PCR |
| armZ-RT-AS | GCCATGCCACAGCCGA | RT-PCR |
| SacI-PA5471.1-armZ-S | CGGGAGCTCTGCGCCGCTCATTTCCATCGCG | Translational fusion |
| BamHI-PA5471.1-armZ-A | CGCGGATCCTCAGTGGTGGTGGTGGTGGTGTCGGCAGCACTCCCCACGGGTC | Translational fusion |
| XhoI-PBAD-(275)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGACCGCAAAGGAGGTTACATCGAT | Translational fusion |
| XhoI-PBAD-(212)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGCGGCCACCGAAACCTACCGCTC | Translational fusion |
| XhoI-PBAD-(86)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGGTTAGACGCCCTTCGCCCTTCCG | Translational fusion |
| XhoI-PBAD-(36)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGCCCCGGATCTACCGTTTCAATCA | Translational fusion |
| XhoI-PBAD-(15)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGCACATGGATTGGATTATGGGCAA | Translational fusion |
| XhoI-PBAD-(pET28)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGGAAGGAGATATACCATGGGCAACTACATCAAGCC | Translational fusion |
| XhoI-PBAD-(36mut)armZ-S | CAACTCTCTACTGTTGGTGGCTCGAGCCCCGGATCTACCGTAAATTAAACATGGATTGGATTATGGGCAAC | Translational fusion |
| XhoI-PBAD-(36)GFP-S | CAACTCTCTACTGTTGGTGGCTCGAGCCCCGGATCTACCGTTTCAATCACATGGATTGGATTATGAGTAAAGGAGAAGAACTTTTCAC | Translational fusion |
| XhoI-PBAD-(15)GFP-S | CAACTCTCTACTGTTGGTGGCTCGAGCACATGGATTGGATTATGAGTAAAGGAGAAGAACTTTTCAC | Translational fusion |
| HindIII-ArmZ/GFP-AS | GTAAAACGACGGCCAGTGCCAAGCTT | Translational fusion |
| EcoRI-PmexZ-S | CCGGAATTCAGCGCTTGAGCTTGTCGG | Transcriptional fusion |
| BamHI-PmexZ-AS | CGCGGATCCGGCGTTTTCGTCGGGTAC | Transcriptional fusion |