Table 1.
Strains and plasmids used in this study
| Strain, bacteriophage, or plasmid | Genotype or relevant characteristicsa | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1Δ(lacZYA-argF)U169[φ80dlacZΔM15]F−NaIr | 38 |
| HB101 | proA2 hsdS20(rB−mB−) recA13 ara-14 lacYI galK2 rpsL20 supE44 xyl-5 mtl-1F− | 38 |
| S17-1/λpir | pro thi hsdR recAchromosome::RP4-2 Tc::Mu Km::Tn7/λpir; TprSmr | 27, 41 |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR176 supE44 relA1 lac[F′ proAB lacIqZΔM15::Tn10(53)] | Stratagene |
| P. aeruginosa | ||
| PAO1 | Wild type | 17 |
| PAO6358 | PAO1 containing a 900-bp deletion in the rpoNlocus | 16 |
| PAO6592 | PAO1 containing a 1,296-bp deletion in the dctAlocus | This study |
| PAO6705 | PAO1 containing a 1,798-bp deletion in the PA5165 (dctB) locus | This study |
| PAO6706 | PAO1 containing a 1,377-bp deletion in the PA5166 (dctD) locus | This study |
| PAO6707 | PAO1 containing a double deletion in the PA5165 locus (1,798 bp) and in the PA5166 locus (1,377 bp) | This study |
| PAO6708 | PAO1 containing a 2,899-bp deletion in the PA5167-PA5169 (dctPQM) operon | This study |
| PAO6709 | PAO1 containing a double deletion in the dctAlocus (1,296 bp) and in the dctPQM(PA5167-PA5169) operon (2,899 bp) | This study |
| PA6710 | PAO6592 with PA5168::Tn5Gm; Gmr | This study |
| PA6815 | PAO1 with PA5168::Tn5Gm; Gmr | This study |
| Bacteriophage | ||
| E79tv-2 | Temperate, transducing variant of E79 | 29 |
| Plasmids | ||
| pLM1 | Tn5Gm delivery vector; GmrApr | 9 |
| pMMB67HE | IncQ expression vector carrying an inducible tacpromoter; Ap/Cbr | 11 |
| pME3087 | Suicide vector for allelic replacement; Tcr; ColE1 replicon | 51 |
| pME6015 | Cloning vector for translational lacZfusions; Tcr | 39 |
| pME6016 | Cloning vector for transcriptional lacZfusions; Tcr | 39 |
| pME9506 | Plasmid carrying a transcriptional dctA-lacZfusion | This study |
| pME9507 | Plasmid carrying a translational dctA′-′lacZfusion | This study |
| pME9508 | Plasmid carrying a translational dctA′-′lacZfusion in which the rpoNputative box of the dctApromoter TGGCAC-N5-CTGCA was replaced with TACCAC-N5-CTTTA | This study |
| pME10031 | Suicide construct used for deletion of the PA5165 gene; Tcr | This study |
| pME10032 | Suicide construct used for deletion of the PA5166 gene; Tcr | This study |
| pME10033 | Suicide construct used for deletion of the PA5167-PA5169 operon; Tcr | This study |
| pME10034 | Plasmid carrying a transcriptional dctP-lacZfusion | This study |
| pME10035 | Plasmid carrying a translational dctP′-′lacZfusion | This study |
| pME10036 | Plasmid carrying a translational dctP′-′lacZfusion in which the rpoNputative box of the dctPQMpromoter TGGCAC-N5-TTGCT was replaced with TACCAC-N5-TTTTT | This study |
| pME10037 | pMMB67HE with dctA | This study |
| pME10038 | pMMB67HE with PA5165 | This study |
| pME10039 | pMMB67HE with PA5166 | This study |
| pME10040 | pMMB67HE with PA5167-PA5169 | This study |
| pME10041 | Suicide construct used for deletion of the dctAgene; Tcr | This study |
a
Nalr, nalidixic acid resistance; Gmr, gentamicin resistance; Tcr, tetracycline resistance; Apr, ampicillin resistance; Smr, streptomycin resistance; Cbr, carbenicillin resistance; Tpr, trimethoprim resistance.