Table 1.
Strains, plasmids, and primers used in the study
| Strain, plasmid, or primer | Relevant characteristics or sequence (5′→ 3′)a | Reference or source |
|---|---|---|
| E. coli | ||
| DH5α | λ− φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | 36 |
| HB101 | proA2hsdS20 (rb−mb−) recA13ara-14 lacY1 galK2 rpsL20 supE44 xyl-5 mtl-1 F− | 36 |
| P. aeruginosa | ||
| PAO1 | Wild type | ATCC 15692 |
| PAO1-retS | PAO1 containing a deletion in the retS gene | 26 |
| PAO6281 | gacA::ΩSm/Sp; Sm/Spr | 35 |
| PAO6327 | gacS::ΩSm/Sp; Sm/Spr | C. Reimmann, unpublished data |
| PAO6733 | PAO1 containing a 8,320-bp deletion in the PA0588-PA0584 genes | This study |
| Plasmids | ||
| pME3087 | Suicide vector for allelic replacement; ColE1 replicon; Tcr | 46 |
| pME3331 | Transcriptional rsmZ-lacZ fusion; Tcr | 15 |
| pME3641 | Plasmid carrying a translational proC′-′lacZ fusion, Cbr | 37 |
| pME3859 | Plasmid carrying a translational rsmA′-′lacZ fusion; Tcr | 33 |
| pME6013 | Broad-host-range vector for translational ′lacZ fusions; Tcr | 14 |
| pME6119 | Translational gacA′-′lacZ fusion; Cbr | 35 |
| pME7311 | Transcriptional rsmY-lacZ fusion; Tcr | 20 |
| pME9902 | Suicide construct derived from pME3087 used for deletion of the PA0588-PA0584 locus; Tcr | This study |
| pME9920 | pME6013 derivative carrying a translational PA0588′-′lacZ fusion; Tcr | This study |
| pRK2013 | Helper plasmid; ColE1 replicon; Tra+ Kmr | 9 |
| Primers | ||
| PRSMZ1 | CGTACAGGGAACACGCAACC | 15 |
| PRSMZ2 | AAAAAAAGGGGCGGGGTATT | 15 |
| PAOTRR1 | GTCAGGACATTGCGCAGGAAGC | 47 |
| PAOTRR2 | AAAACCCCGCCTTTTGGGCG | 47 |
| UPPA0588 | CGAAAGCTTCACCCGCATGCGT | This study |
| PA0588-78 | GCTGACGTCGAGATATTCCTGGAG | This study |
| PA0584-1226 | AGACTGCAGGCCTGAAGCAGCGC | This study |
| DOWNPA0584 | TATGAATTCGCCCGGGAATC | This study |
| TranslfusPA0588-F | CAGGGATCCTATTTCATCCAGCAG | This study |
| TranslfusPA0588-R | TCCCTGCAGAAAATGCTCATGACGTC | This study |
| 5SrRNA-1 | GCCTAAGCTTTTGGACAGGATGGGGTTGGA | 13 |
| 5SrRNA-2 | CAAAGAATTCGACGATTGTGTGTTGTAAGG | 13 |
a
Underlined sequences in primers indicate restriction sites as specified in Materials and Methods.