TABLE 1.
Strains, plasmids, and oligonucleotides used in the current study
| Strain, plasmid, or oligonucleotide | Relevant characteristic(s) or sequence | Reference or source |
|---|---|---|
| Strains | ||
| Pseudomonas aeruginosa | ||
| PAO1 | Wild type | 23 |
| PW9687 | PA5166-F04::ISphoA/hah derivative of PAO1 | 24 |
| PW10371 | PA5530-C01::ISphoA/hah derivative of PAO1 | 24 |
| PAO6359 | rpoN::Ω-Km derivative of PAO1 | 25 |
| ΔmifR PAO1 | ΔmifR derivative of PAO1 | This study |
| ΔmifR PW9687 | ΔmifR derivative of PW9687 | This study |
| PA14 | Wild type | 26 |
| ΔPA5530 PA14 | ΔPA5530 derivative of PA14 | This study |
| PAK | Wild type | 27 |
| ΔPA5530 PAK | ΔPA5530 derivative of PAK | This study |
| Escherichia coli Top10 | F− mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL (Strr) endA1 λ− | Invitrogen |
| Plasmids | ||
| pCR-Blunt | Cloning plasmid; Kmr | Invitrogen |
| pTrc99a | Expression plasmid; Cbr | Pharmacia |
| pBBR1MCS-5 | Broad-host-range plasmid; Gmr | 28 |
| pEX18ApGW | Plasmid for gene deletions in P. aeruingosa; Cbr Gmr | 29 |
| ΔPlac-pBBR1MCS-5 | pBBR1MCS-5 minus lac promoter; Gmr | This study |
| pBRL474 | PA5530 gene in pCR-Blunt; Kmr | This study |
| pBRL479 | PA5530 gene in pBBR1MCS-5; Gmr | This study |
| pBRL476 | PA5530-lacZ in pCR-Blunt; Kmr | This study |
| pBRL485 | PA5530-lacZ in ΔPlac-pBBR1MCS-5; Gmr | This study |
| pBRL487 | PA5530-lacZ with mutated RpoN promoter; Gmr | This study |
| pBRL511 | mifR::Gmr in pEX18ApGW; Cbr Gmr | This study |
| pBRL538 | PA5530::Gmr in pEXT18ApGW; Cbr Gmr | This study |
| pBRL549 | mifSR operon in pCR-Blunt; Kmr | This study |
| pBRL561 | mifS-ΔmifR operon in pCR-Blunt; Kmr | This study |
| pBRL566 | mifSR operon (forward orientation) in pTrc99a; Cbr | This study |
| pBRL567 | mifSR operon (backward orientation) in pTrc99a; Cbr | This study |
| pBRL570 | mifS-ΔmifR operon (forward orientation) in pTrc99a;Cbr | This study |
| pJRH07 | mifR gene in pCR-Blunt; Kmr | This study |
| pJRH08 | mifR gene in pBBR1MCS-5; Gmr | This study |
| Oligonucleotides | ||
| BL342.f | ATGACCATGATTACGGATTCACTG | |
| BL342.r | GCAGTTATTTTTGACACCAGACCAACTGGTA | |
| BL434.f | GCATCTAGACTGATCAGCACATCCAAGACAAC | |
| BL434.r | GCAGAGCTCTCAATCGGTCGTGATCTTCGA | |
| BL435.f | GCAGATCGGCGAGTTCTCC | |
| BL435.r | GAATCCGTAATCATGGTCATCGTGTTTCCTCTTTTCGTTGTTG | |
| BL438.f | GCTTTCCCGCGTTAACACGGCACCTGCTATC | |
| BL438.r | GATAGCAGGTGCCGTGTTAACGCGGGAAAGC | |
| BL466.f | TACAAAAAAGCAGGCTGACCAGGTGATCTTCGTCGAC | |
| BL466.r | TCAGAGCGCTTTTGAAGCTAATTCGGAAGGGTTTCTCGATGAAGTCG | |
| BL467.f | AGGAACTTCAAGATCCCCAATTCGCGAACATTTCGCCAGCGTTG | |
| BL467.r | TACAAGAAAGCTGGGTCTCTTCGCCGACGAAATCGC | |
| BL552.f | TACAAAAAAGCAGGCTGAAAGCGCCAACGCCATTTC | |
| BL552.r | TCAGAGCGCTTTTGAAGCTAATTCGGATGATCAGCGAGCCAAAGC | |
| BL553.f | AGGAACTTCAAGATCCCCAATTCGCTTCTACACCTACACCACCTAC | |
| BL553.r | TACAAGAAAGCTGGGTCAATCGGTCGTGATCTTCGAG | |
| BL554.f | GCATCTAGACGTTTCGATCCGCCGATGTC | |
| BL554.r | GCAGAGCTCAGTTGGCGAAGGATCTCTGAC | |
| JRH04.f | GACTCTAGATAAGAAGGAGATATACCATGAGCGACCAGGTGATCTTC | |
| JRH04.r | GACCTCGAGAGTTGGCGAAGGATCTCTGAC |