Table 1.
Bacterial strains and plasmids used in this work
| Strains/Plasmid | Relevant characteristicsa | Source/Reference |
|---|---|---|
| Strain | ||
| Escherichia coli | ||
| TG1 | supEΔ(lac-proAB) thi hsdRΔ5 (F' traD36 rpoA+B+lacIqZΔM15) | Lab collection |
| SM10 (λpir) | thi-1 thr-1 leuB6 supE44 tonA21 lacY1 recA-::RP4-2-Tc::Mu Kmr | Lab collection |
| TOP10F' | F'[lacIq Tn10(tetR)]mcrAΔ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 deoR nupG recA1 araD139Δ (ara-leu)7697 galU galK rpsL (Strr) endA1λ | Invitrogen |
| CC118 (λpir) | Host strain for pKNG101 replication; Δ(ara-leu) araDΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 Rfr (λpir) | Lab collection |
| DHMI | cya-854 recA1 gyrA96 (NaI) thi1 hsdR17 spoT1 rfbD1 glnV44(AS) | Karimova et al. (1998) |
| Pseudomonas aeruginosa | ||
| PAK | Wild type | D. Bradley |
| PAKΔrocA1 | In frame deletion of rocA1 (PA3948) in PAK | Kulasekara et al. (2005) |
| PAKΔrocR | In frame deletion of rocR (PA3947) in PAK | Kulasekara et al. (2005) |
| PAKΔrocA2 | In frame deletion of rocA2 (PA3045) in PAK | This study |
| PAKΔrocA1-rocA2 | In frame deletion of rocA1 (PA3948) and rocA2 (PA3045) in PAK | This study |
| PAK::cupC-lacZ | PAK with a cupC-lacZ fusion integrated on the chromosome at the attB site | This study |
| PAKΔrocA1::cupC-lacZ | rocA1 deletion mutant in PAK::cupC-lacZ | This study |
| PAKΔrocA2::cupC-lacZ | rocA2 deletion mutant in PAK::cupC-lacZ | This study |
| PAKΔrocR::cupC-lacZ | rocR deletion mutant in PAK::cupC-lacZ | This study |
| PAK::cupB-lacZ | PAK with a cupB-lacZ fusion integrated on the chromosome at the ctx att site | This study |
| PAKΔrocA1::cupB-lacZ | rocA1 deletion mutant in PAK::cupB-lacZ | This study |
| PAKΔrocA2::cupB-lacZ | rocA2 deletion mutant in PAK::cupB-lacZ | This study |
| PAKΔrocA1-rocA2::cupB-lacZ | rocA1-rocA2 double deletion mutant in PAK::cupB-lacZ | This study |
| Plasmid | ||
| pRK2013 | ColE1 mob+traRK2ΔrepRK2repE- Kmr | Figurski and Helinski (1979) |
| pCR2.1 | TA cloning vector for PCR products, lacZα ColE1 f1 ori Apr Kmr | Invitrogen |
| pMMB67HE | Broad host range vector, tac promoter Apr | Lab collection |
| pMMB67EH-GW | Broad host range vector, tac promoter Gmr | Lab collection |
| pET-Dest42 | Destination vector for gateway technology, T7 promoter, Apr | Invitrogen |
| pMMB67HE42 | pMMB67HE containing the gateway cassette from pET-Dest42 Apr | E. Termine |
| pMMB67-rocS2 | pMMB67HE harbouring the rocS2 gene from the Gateway library | This study |
| pMMB67-rocA2 | pMMB67HE harbouring the rocA2 gene from the Gateway library | This study |
| pMMB67-rocS1 | pMMB67EH-GW harbouring the rocS1 gene from the Gateway library | Kulasekara et al. (2005) |
| pKNG101 | Suicide vector for P. aeruginosa. sacB, Smr | Kaniga et al. (1991) |
| pKNGΔrocA2 | pKNG101 containing 560 bp upstream and 505 bp downstream from the rocA2 gene DNA fragment | This study |
| miniCTX-lacZ | Vector for unmarked integration into P. aeruginosa att site. oriT, FRT, int, Tcr | Hoang et al. (2000) |
| miniCTX-cupC-lacZ | cupC1 promoter cloned into miniCTX-lacZ (BamHI/XhoI) | This study |
| miniCTX-cupB-lacZ | cupB1 promoter cloned into miniCTX-lacZ (BamHI/XhoI) | This study |
| pKT25 | Cloning and expression vector, encodes the T25 fragment (amino acids 1–224 of CyaA), Kmr | Karimova et al. (1998) |
| pUT18c | Cloning and expression vector, encodes the T18 fragment (amino acids 225–399 of CyaA), Apr | Karimova et al. (1998) |
| pKT25-torR | DNA fragment encoding the D2 domain of TorR cloned into pKT25 | Kulasekara et al. (2005) |
| pUT18c-torS | DNA fragment encoding the Hpt domain of TorS cloned into pUT18c | Kulasekara et al. (2005) |
| pKT25-rocS1 | DNA fragment encoding the Hpt domain of RocS1 cloned into pKT25 | Kulasekara et al. (2005) |
| pUT18c-rocA1 | DNA fragment encoding the D2 domain of RocA1 cloned into pUT18c | Kulasekara et al. (2005) |
| pUT18c-rocR | DNA fragment encoding the D2 domain of RocR cloned into pUT18c | Kulasekara et al. (2005) |
| pKT25-rocS2 | DNA fragment encoding the Hpt domain of RocS2 cloned into pKT25 | This study |
| pUT18c-rocA2 | DNA fragment encoding the D2 domain of RocA2 cloned into pUT18c | This study |
| pUT18c-trpO | DNA fragment encoding the D2 domain of PA0034 cloned into pUT18c | This study |
| pKT25-gacS | DNA fragment encoding the H1-D1-Hpt domain of GacS cloned into pKT25 | This study |
| pUT18c-gacA | DNA fragment encoding the D2 domain of GacA cloned into pUT18c | This study |
a
Apr, ampicillin resistance; Smr, Streptomycin resistance; Kmr, Kanamycin resistance, Tcr, Tetracycline, Gmr, Gentamicin resistance.