TABLE 3.
Strains and plasmids used in this studya
| Strain or plasmid | Descriptionb | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5αMCR | Strain used for general cloning | Life Technologies (Grand Island, NY) |
| S17-1 λ pir | Strain used for conjugation | 66 |
| F. columnare strains | ||
| MS-FC-4 | Wild type | 31, 32 |
| FCB 115 | ΔsidL, siderophore synthesis region (C6N29_09920, C6N29_09925, C6N29_09930, C6N29_09935, C6N29_09940, C6N29_09945, C6N29_09950, C6N29_09955, C6N29_09960, C6N29_09965, C6N29_09970) | This study |
| FCB161 | ΔsidL, ΔiucX | This study |
| FCB164 | ΔiucX | This study |
| FCB166 | ΔiucA | This study |
| FCB175 | ΔiucX, ΔiucA | This study |
| FCB199 | ΔiucX, ΔiucA, ΔiucC | This study |
| FCB208 | ΔiucC | This study |
| FCB219 | ΔiucA, ΔiucC | This study |
| FCB231 | ΔsidL, ΔiucX complemented by iucX inserted into the chromosome | This study |
| FCB232 | ΔiucX, ΔiucA, ΔiucC complemented by iucA inserted into the chromosome | This study |
| Plasmids | ||
| pCP23 | E. coli-F. columnare shuttle plasmid; Apr (Tcr) | 67 |
| pMS75 | Suicide vector carrying sacB; Apr (Tcr) | 10 |
| pRC44 | 2.2-kbp region downstream of the siderophore biosynthesis genes amplified using primers 2465 and 2466 and cloned into KpnI and BamHI sites of pMS75; Apr (Tcr) | This study |
| pRC45 | 2.1-kbp region upstream of the siderophore biosynthesis genes amplified using primers 2467 and 2468 and cloned into BamHI and SalI sites of RC44; Apr (Tcr) | This study |
| pRC48 | 1.9-kbp fragment containing iucA amplified using primers 2546 and 2547 and cloned into KpnI and PstI sites of pCP23; Apr (Tcr) | This study |
| pRC49 | 2.2-kbp region downstream of iucA amplified using primers 2542 and 2543 and cloned into KpnI and BamHI sites pMS75; Apr (Tcr) | This study |
| pRC50 | 2.0-kbp region upstream of iucA amplified using primers 2544 and 2545 and cloned into BamHI and SalI sites of pRC49; Apr (Tcr) | This study |
| pRC56 | 1.8-kbp fragment containing iucX amplified using primers 2564 and 2565 and cloned into KpnI and PstI sites of pCP23; Apr (Tcr) | This study |
| pRC57 | 2.2-kbp region downstream of iucX amplified using primers 2560 and 2561 and cloned into BamHI and SalI sites of pMS75; Apr (Tcr) | This study |
| pRC58 | 2.6-kbp region upstream of iucX amplified using primers 2562 and 2563 and cloned into SalI and SphI sites of pRC57; Apr (Tcr) | This study |
| pRC62 | 2.7-kbp fragment containing iucC amplified using primers 2587 and 2588 and cloned into KpnI and PstI sites of pCP23; Apr (Tcr) | This study |
| pRC63 | 2.1-kbp region upstream of iucC amplified using primers 2585 and 2586 and cloned into BamHI and SalI sites of pMS75; Apr (Tcr) | This study |
| pRC64 | 2.3-kbp region downstream of iucC amplified using primers 2583 and 2584 and cloned into KpnI and BamHI sites of pRC63; used to delete iucC from FCB166 and FCB175, generating FCB219 and FCB232; Apr (Tcr) | This study |
| pRC67 | 2.1-kbp region downstream of iucC amplified using primers 2599 and 2600 and cloned into KpnI and BamHI sites of pRC63; used to delete iucC, generating FCB208; Apr (Tcr) | This study |
| pRC69 | 6.3-kbp region containing upstream, iucX, and downstream amplified using primers 2560 and 2563 and cloned into BamHI and SphI sites of pMS75; Apr (Tcr) | This study |
| pRC71 | 6.8-kbp region containing upstream, iucA, and downstream amplified using primers 2601 and 2586 and cloned into BamHI and SalI sites of pMS75; Apr (Tcr) | This study |
a
kbp, kilobase pair.
b
Antibiotic resistance phenotypes: ampicillin (Apr) and tetracycline (Tcr). Unless indicated otherwise, the antibiotic resistance phenotypes are those expressed in E. coli. The antibiotic resistance phenotypes given in parentheses are those expressed in F. columnare but not in E. coli.