TABLE 1.
Strains and plasmids used in this study
| Plasmid or strain | Description | Reference |
|---|---|---|
| pSUP202 | pBR325 containing RP4 Mob site; Cmr Tcr Apr | 24 |
| pUC4K | pUC4 with aminoglycoside 3′-phosphotransferase gene from Tn903 (Kmr) | Pharmacia Biotech |
| λFAJ2001 | λEMBL3 with Sau3AI fragment carrying P. fluorescens OE 28.3 oprF gene | 1 |
| pFAJ2001 | pCL1921 with 3.3-kb SmaI fragment of λFAJ2001 carrying oprF gene in SmaI site | 1 |
| pCL1821 | Low-copy-number E. coli plasmid; Spr | 12 |
| pFAJ2052 | pCL1921 with 3.3-kb SmaI fragment of λFAJ2001 carrying oprF gene in HindIII site | This work |
| pFAJ2073 | pFAJ2052 with 1.26-kb HindIII fragment of pUC4K conferring Kmr inserted in NruI site of oprF | This work |
| pFAJ2082 | pSUP202 with BamHI-HindIII fragment of pFAJ2073 containing disrupted oprF | This work |
| pFAJ2425 | pFAJ2052 with 1.26-kb HindII fragment of pUC4K inserted in Asp700 site of sigX | This work |
| pFAJ2431 | pSUP202 with BamHI-HindIII fragment of pFAJ2425 containing disrupted sigX | This work |
| pFAJ2432 | pUC19 with PCR fragment containing P. fluorescens M114 sigX gene in BamHI site | This work |
| pFAJ2448 | pUC18 with PCR fragment containing P. syringae pv. syringae sigX gene, in BamHI site | This work |
| pFAJ2511 | pUC18 with 1.5-kb SacI fragment of λFAJ2001 (upstream of sigX) | This work |
| pFAJ2672 | pUC18 with 4.8-kb XhoI fragment of λFAJ2001 (upstream of sigX) | This work |
| pRW5 | pUCP19 + 1.47-kb HindIII-EcoRI fragment coding for P. aeruginosa H103 OprF | 27a |
| pEX100t | pUC19-based gene replacement vector with a counterselectable sacB marker | 23 |
| pX1918GT | Contains xylE-Gmr cassette (SmaI fragment) for construction of gene disruptions and reporter gene fusions | 23 |
| pWW2300 | pUC19 + 1.3-kb PstI-SalI insert (contains 3′ end of cmpX, sigX, and the 5′ end of oprF from P. aeruginosa H103) | 28 |
| pWW1701 | pUC19 + 1.7-kb SmaI insert (contains cmpX, sigX, and the 5′ end of oprF from P. aeruginosa H103) | 28 |
| pWW1901 | pUC19 + 1.9-kb SalI insert (contains cmaX, crfX, cmpX, sigX, and the 5′ end of oprF from P. aeruginosa H103) | 28 |
| pFB2e1 | xylE-Gmr cassette (SmaI fragment) inserted into the EcoRV site within P. aeruginosa sigX in pWW1701 | This work |
| pFB2e1a3 | 3.4-kb FspI-SmaI fragment of pFB2e1 (xylE-Gmr cassette in SigX) cloned into Sma I site of pEX100t | This work |
| E. coli | ||
| CL83 | recA mutant (recA56) derivative of JM83 | 12 |
| DH5α | Strain used for all standard cloning experiments | |
| S17-1 | Mobilizing strain for RP4 Mob-containing plasmids | 24 |
| P. aeruginosa | ||
| H103 | PAO1 prototroph; wild-type reference strain | 9 |
| H636 | oprF::Strr mutant of P. aeruginosa H103 | 29 |
| H814 | sigX::Gmr mutant of P. aeruginosa H103 | This work |
| H845 | P. aeruginosa H814 + pRW5 (sigX::Gmr mutant with OprF expressed from a plasmid) | This work |
| P. fluorescens | ||
| FAJ2026 | oprF mutant of P. fluorescens OE 28.3 | This work |
| FAJ2030 | sigX mutant of P. fluorescens OE 28.3 | This work |
| M114 | Isolated from sugarbeet rhizosphere | 7 |
| OE 28.3 | Isolated from wheat rhizosphere | 4 |
| P. syringae LMG 1247T | Isolated from Syringa vulgaris; pathovar reference strain |