TABLE 1.
Bacterial strains and plasmids used in this study
| Strains, vector, or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| V. vulnificus | ||
| C7184/OP | Wild type (clinical isolate) | 73 |
| C7184DΩ | C7184 with disrupted pilD | This laboratory |
| C7184AΩ | C7184 with disrupted pilA | This study |
| MO6-24/OP | Wild type (clinical isolate) | 34 |
| PAC1 | Wild type (environmental isolate) | M. Coyle |
| P. aeruginosa | ||
| PAK-NP | Nonpiliated mutant | 46 |
| PAK-NP(pRP383) | Nonpiliated mutant expressing PilA from V. vulnificus | This study |
| E. coli | ||
| DH5α | supE44lacU169(ΔlacZΔM15)hsdR17 recA lendA1gyrA96 thi-1 relA1 | BRL |
| SM10-λpir | thi thr leu tonAlacY supE recA[RP4-2-Tc::Mu] λpirR6K Kmr | 51 |
| BL21 | F−ompT hsdSB (rB− mB−)gal dcm | 61, 62 |
| pFLAG-MAC | Apr FLAG fusion cloning vector | Sigma |
| pFLAG-ATS | Apr FLAG fusion cloning vector | Sigma |
| Cloning vectors | ||
| pZErO-2 | Kanr Zero background cloning vector | Invitrogen |
| pMMB67EH.cam | Cmr Apr broad-host-range cloning vector, lacIq/tac promoter | 17 |
| pBluescript II KS+ | Apr phagemid cloning vector | Stratagene |
| pEP185.2 | Cmr suicide vector | 22 |
| Recombinant plasmids | ||
| pRP-P38 | 9- to 10-kb PstI fragment from C7184 carrying the pilABCD cluster cloned into pZErO-2 | This study |
| pRP102 | 2.04-kb PCR-generated fragment encompassing pilA cloned into pKS+ | This study |
| pRP102Ω | 2.04-kb PCR-generated fragment encompassing pilA, disrupted with the 2.0-kb Ω interposon, cloned into pEP185.2 | This study |
| pRP383 | 624-bp XhoI-HindIII fragment encompassing pilA cloned into pMMB67EH.cam | This study |
| pRPA110 | PCR-generated pilA cloned into pFLAG-MAC | This study |
| pRPA111 | PCR-generated pilA cloned into pFLAG-ATS | This study |