Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or phenotype | Reference |
|---|---|---|
| P. aeruginosa strains | ||
| ATCC 17933 | Wild type | 29 |
| MD1 | ATCC 17933 derivative, ΔPA3604::Kmr | 8 |
| NH1 | ATCC 17933 derivative, PA1991::Kmr | This study |
| NH2 | SH1 derivative, PA1991::Kmr ΔPA1992::Gmr | This study |
| NH3 | ATCC 17933 derivative, ΔPA2572::Kmr | This study |
| NH5 | ATCC 17933 derivative, PA5186::Kmr | This study |
| SH1 | ATCC 17933 derivative, ΔPA1992::Gmr | 8 |
| E. coli strains | ||
| DH5α | supE44 ΔlacU169(ϕ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 30 |
| HB101 | supE44 hsdS20(rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 31 |
| Plasmids | ||
| pACΩGm | Gmr; cloning vector containing Gmr cassette and Ω fragments | 32 |
| pEDY305 | Tcr; lacZ promoter-probe vector | 22 |
| pEX18Ap | Apr; oriT+ sacB+, gene replacement vector with multiple cloning site (MCS) from pUC18 | 20 |
| pLAFR3 | Tcr; broad-host-range cosmid | 33 |
| pQF50 | Apr; lacZ promoter-probe vector | 21 |
| pRK2013 | Kmr; helper plasmid for triparental mating | 34 |
| pUC18, 19 | Apr; cloning and expression vector | 35 |
| pUCP20T | Apr; broad-host-range vector | 36 |
| pTB3001 | Tcr; ∼25-kbp genomic DNA partially digested with Sau3AI from P. aeruginosa cloned into BamHI site of pLAFR3 | 6 |
| pTB3070 | Apr; 3.2-kbp BamHI-PstI fragment from pTB3001 cloned into BamHI-PstI sites of pUC18 | 6 |
| pTB3112 | Apr; 6.2-kbp PstI-PstI fragment from pTB3001 cloned into PstI site of pUCP20T | 3 |
| pTB3131 | Apr Kmr; 1.0-kbp PCR product with promoter and Kmr cassette of Tn5 cloned into EcoRI-BamHI sites of pUC19 | 16 |
| pTB3135 | Apr; 0.89-kbp HindIII-SalI-fragment from pTB3070 cloned into HindIII-SalI sites of pUC19 | 3 |
| pTB3138 | Tcr; P_exaA′-lacZ; 0.89-kbp PstI-SalI fragment from pTB3070 cloned into PstI-XhoI sites of pEDY305 | 3 |
| pTB3139 | Tcr; P_exaB′-lacZ; 0.89-kbp PstI-XbaI fragment from pTB3135 cloned into PstI-XbaI sites of pEDY305 | 3 |
| pTB3144 | Apr; 3-kbp MluI-SmaI fragment of pTB3112 removed, ends filled in, and vector religated | 3 |
| pTB7023 | Tcr; P_pqqAB́-lacZ; 2.5-kbp XhoI-BamHI fragment of pTB3070 cloned into XhoI-BglII sites of pEDY305 | 5 |
| pTB7060 | Apr; 1.67-kbp PCR product of erbR cloned into PstI-BamHI sites of pUCP20T | 5 |
| pTB7074 | Apr; P_eraSR(PA1979/1980)′-lacZ; 1.16-kbp promoter region of exaDE cloned into BglII-XbaI sites of pQF50 | 5 |
| pTB7082 | Apr; P_ercA(PA1991)′-lacZ; 0.53-kbp PCR product containing promoter region of PA1991 cloned into XbaI-HindIII sites of pQF50(pPA1991) | 8 |
| pTB7101 | Apr; 1.98-kbp PCR product of PA1992 cloned into SphRI-EcoRI sites of pUCP20T | 8 |
| pTB7107 | Apr; P_erdR(PA3604)′-lacZ; 280-bp promoter region of PA3604 cloned into XbaI-HindIII sites of pQF50 | 8 |
| pTB7108 | Apr; P_erbR(PA3604)′-lacZ; 1.0-kbp promoter region of agmR cloned into KpnI-HindIII sites of pQF50 | 8 |
| pTB7200 | Apr; 3.3-kbp PCR product containing ercA-ercS and its promoter cloned into EcoRI-HindIIII sites of pUCP20T | This study |
| pTB7212 | Apr; 1.2-kbp PCR product of PA1991 cloned into EcoRI-HindIII sites of pUCP20T | This study |
| pTB7219 | Apr; 1.2-kbp PCR product of PA1146 cloned into EcoRI-HindIII sites of pEX18Ap | This study |
| pTB7220 | Apr; 1.2-kbp PCR product of PA1991 cloned into EcoRI-HindIIII sites of pEX18Ap | This study |
| pTB7222 | Apr; 1.3-kbp PCR product of PA2572 cloned into EcoRI-HindIII sites of pEX18Ap | This study |
| pTB7224 | Apr; 1.2-kbp PCR product of PA5186 cloned into EcoRI-KpnI sites of pEX18Ap | This study |
| pTB7225 | Apr Kmr; 380-bp SmaI-SmaI fragment of pTB7219 removed and filled with PCR product of Kmr cassette | This study |
| pTB7226 | Apr Kmr; 1.0-kbp PCR product of Kmr cassette cloned into SmaI site of pTB7220 | This study |
| pTB7227 | Apr Kmr; 500-bp PstI-PstI fragment of pTB7222 removed and filled with PCR product of Kmr cassette | This study |
| pTB7229 | Apr Kmr; 1.0-kbp PCR product of Kmr cassette cloned into blunted NcoI site of pTB7224 | This study |
| pTB7234 | Apr; P_PA1146́-lacZ; 500-bp PCR product containing promoter region of PA1146 cloned into NcoI-HindIIII sites of pQF50(pPA1146) | This study |
| pTB7235 | Apr; P_PA2572′-lacZ; 500-bp PCR product containing promoter region of PA2572 cloned into NcoI-HindIIII sites of pQF50 | This study |
| pTB7237 | Apr; P_PA5186′-lacZ; 500-bp PCR product of promoter region of PA5186 cloned into NcoI-HindIIII sites of pQF50(pPA5186) | This study |