TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| V. anguillarum H-775-3 | Serotype O1 plasmidless avirulent strain, deficient in anguibactin biosynthesis, derived from strain 775 by curing of pJM1 | J. H. Crosa |
| E. coli | ||
| DH5α | Cloning strain | Laboratory stock |
| SM17-1-λpir | Cloning strain | Laboratory stock |
| EB53 | aroB hemA | 6 |
| 101ESD | HB101 derivative, deficient in enterobactin biosynthesis Δ(entC-entA) | J. H. Crosa |
| Plasmids | ||
| pGEMT-Easy | PCR cloning vector, Apr | Promega |
| pWKS30 | Low-copy-number plasmid vector, Apr | 41 |
| pACYC177 | Cloning vector, Apr Kmr | 33 |
| pACYC184 | Cloning vector, Apr Cmr | 33 |
| pVK100 | Mobilizable cloning vector, Tcr Kmr | 15 |
| pRK2013 | Mobilizing plasmid for triparental mating, Kmr | 5 |
| pKEK229 | Suicide vector derived from pCVD442, Apr | 3 |
| pNidKan | pKEK229 with a 1.5-kb PstI-PstI fragment containing the Km cassette from pKANπ | This study |
| pML1 | 21-kb fragment from a gene library of strain H775-3 cloned into pVK100, Tcr | 25 |
| pCAR121 | ApaL1 fragment of pML1 containing huvA gene cloned into pACYC177; Kmr | This study |
| pCAR126 | ApaL1 fragment of pML1 containing huvX, tonB-exbBD, and huvB genes and partial huvZ and huvC genes cloned into pACYC177, Kmr | This study |
| pCAR115 | PCR-amplified huvA gene cloned into pWKS30, Apr | This study |
| pCAR179 | huvZ gene cloned in pWKS30, Apr | This study |
| pCAR181 | huvX gene cloned in pWKS30, Apr | This study |
| pSML11 | EcoRV fragment of pML1 containing complete huvZX genes cloned into pWKS30 | This study |
| pSML23 | 9-kb PCR fragment encoding the heme uptake system of V. anguillarum H-775-3 cloned into pWKS30, Apr | This study |
| pSML32 | PstI-EcoRI fragment of pSML23 containing huvBCD genes cloned into pWKS30, Apr | This study |
| pSML33 | SacI-ApaI fragment of pSML23 containing tonB-exbBD and huvBCD genes cloned into pWKS30, Apr | This study |
| pSML34 | SacI-HindIII fragment of pML1 containing tonB-exbBD genes cloned into pWKS30, Apr | This study |