TABLE 1.
Bacterial strains, plasmids and phage used
| Strain, plasmid, or phage | Relevant characteristics | Source or reference(s) |
|---|---|---|
| V. vulnificus | ||
| MO6-24/OP | Wild type (clinical isolate) | J. D. Oliver |
| C7184/OP | Wild type (clinical isolate) | J. D. Oliver |
| PAC1 | Wild type (environmental isolate) | M. Coyle |
| C7184-D12Ω | C7184 with disrupted vvpD | This study |
| C7184-402K | C7184 with disrupted vvhA | This laboratory |
| E. coli | ||
| DH5α | supE44 lacU169(φlacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Bethesda Research Laboratories |
| S17-1λpir | supE44 hsdR endA1 recA thi pro RP4-2-Tc::Mu-kan::Tn7 (λpir) | 32, 50 |
| P. aeruginosa | ||
| PAK | Wild type | S. Lory |
| PAK 2B18 | pilD::Tn5 | S. Lory (36) |
| PAK-CΩ | PAK mutant with the Ω interposon inserted in the pilC gene | 23 |
| Cloning vectors | ||
| pLAFR3 | Tcr broad-host-range plasmid vector | 52 |
| pBluescript II SK+ | Apr phagemid cloning vector | Stratagene |
| pMMB67HE.cam | Cmr Apr broad-host-range cloning vector, lacIq/tac promoter | 17 |
| pEP185.2 | Cmr suicide vector | 20 |
| pZErO-2 | Kanr Zero background cloning vector | Invitrogen |
| Recombinant plasmids | ||
| pJCL12B | 25-kb Sau3AI fragment from V. vulnificus MO6-24 in pLAFR3 | This study |
| pMS450 | 4.8-kb PstI fragment from pJCL12B cloned into pLAFR3 (vvpBCD) | This study |
| pMS451 | 2.8-kb HindIII-PstI fragment cloned into pLAFR3 (vvpCD) | This study |
| pMS4514 | 2.8-kb HindIII-PstI fragment cloned into pBluescript II SK+ | This study |
| pMS451Ω | 2.8-kb HindIII-PstI fragment with Ω fragment inserted in the NarI site of vvpD | This study |
| pMS451Ω-2 | 4.8-kb KpnI-SmaI fragment from pMS451Ω in pEP185.2 | This study |
| pRPD1 | 1.0-kb SalI fragment from C7184 cloned into pMMB67HE.cam (vvpD) | This study |
| Bacteriophage PO4 | P. aeruginosa pilus-specific phage | S. Lory |