TABLE 1.
Strains and plasmids
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Escherichia coli strains | ||
| DH5α | supE44 ΔlacU169(φ80lacZΔM15) hsdR17 thi-1 relA1 recA1 | 15 |
| S17-1(λpir) | thi pro hsdR hsdM+ ΔrecA RP4-2::TcMu-Km::Tn7 | 37 |
| Top10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR nupG recA1 araD139 Δ(ara-leu)7697 galU galK rpsL(Strr) endA1 | Invitrogen |
| Pseudomonas fluorescens strains | ||
| Pf0-1 | Wild type | 9 |
| ΔlapD | Clean deletion of lapD in Pf0-1 | 27 |
| ΔlapD apaH | ΔlapD::pKO-apaH Tcr | This study |
| lapA | Pf0-1::pUC-lapA Kmr | This study |
| lapA apaH | Pf0-1::pUC-lapA::pKO-apaH Tcr Kmr | This study |
| Δpst | Pf0-1 with deletion of pstSCAB-phoU; Gmr | 25 |
| Δpst apaH | Δpst::pKO-apaH Tcr Gmr Tcr | 25 |
| Pfl_5137/apaH | Pf0-1::pKO-apaH Tcr | 25 |
| PF-B337 | Pf0-1 Pfl_5137::mini-Tn5 lacZ1(Km) Kmr | 25 |
| PF-013 | Pf0-1 lapA-HA | 24 |
| PF-201 | Pf0-1 lapA-HA::pKO-apaH | This study |
| PF-001 | Pf0-1::pUC-PlapA-luc Kmr | 24 |
| PF-203 | apaH::pUC-PlapA-luc Kmr Tcr | This study |
| PF-204 | Pf0-1::pUC-PphoX-luc Kmr | 25 |
| PF-205 | apaH::pUC-PphoX-luc Kmr Tcr | This study |
| PF-206 | Pf0-1::Tn7-Ppst-luc Kmr | This study |
| PF-207 | apaH::Tn7-Ppst-luc Kmr Tcr | This study |
| PF-208 | Pf0-1::pUC-PrplU-luc Kmr | This study |
| PF-209 | apaH::pUC-PrplU-luc Kmr Tcr | This study |
| PF-004 | Pf0-1::pUC-PlapE-luc Kmr | 24 |
| PF-210 | apaH::pUC-PlapE-luc Kmr Tcr | This study |
| PF-211 | PF-013 Plac::lapA | This study |
| Plasmids | ||
| pBBRMCS-2 | Broad-host-range cloning vector; Kmr | 21 |
| pBB-apaH (pBB-5137) | pBBRMCS-2 expressing apaH (Pfl_5137 ORF) | 25 |
| pBB-lapA | lapA expression vector for Pseudomonas; Kmr | This study |
| pHRB2 | pUC18T-Tn7T containing Km gene from pZE21-MCS2; Kmr | 25 |
| pKO3 | Pseudomonas integration vector; multiple cloning site, oriT lacZ′; Tcr | 25 |
| pLapA | Clone of lapA ORF; Gmr | This study |
| pMQ71B | pMQ71 modified to remove aacC1; Knr | 25 |
| pMQ72 | Pseudomonas expression vector; Gmr | 36 |
| pLapEBC | lapEBC genes expressed via PBAD promoter | 24 |
| pMQ-His-apaH | pMQ72 expressing N-terminal His-tagged apaH | This study |
| pMQ-Plac-lapA | Vector used to place Plac promoter in front of lapA ORF; Gmr | This study |
| pTn7-Ppst-luc | Tn7 delivery vector for Ppst luciferase fusion; Apr Kmr | This study |
| pUC18T-Tn7T | Mini-Tn7 vector; Apr | 8 |
| pUC-lucK | Vector used for construction of luciferase transcription fusions; Kmr | 25 |
| pUC-PlapA-luc | pUC-lucK with PlapA luciferase fusion; Kmr | 24 |
| pUC-PlapE-luc | pUC-lucK with PlapE luciferase fusion; Kmr | 24 |
| pUC-PphoX-luc | pUC-lucK with PphoX luciferase fusion; Kmr | 25 |
| pUC-PrapA-luc | pUC-lucK with PrapA luciferase fusion; Kmr | This study |
| pUC-PrplU-luc | pUC-lucK with PrplU luciferase fusion; Kmr | This study |
| pUX-BF13 | Tn7 helper plasmid encoding Tn7 transposition functions; Apr | 39 |