TABLE 1.
Plasmids and bacterial strains used in this study
| Strain or plasmid | Relevant characteristicsa | Ref. or source |
|---|---|---|
| Bacterial strains | ||
| V. vulnificus | ||
| MO6-24/O | Wild type, clinical isolate; virulent | 61 |
| MORR | MO6-24/O with spontaneous Rifr mutation, virulent | Laboratory collection |
| MORSR | MO6-24/O with spontaneous Rifr, Smr mutation, virulent | Laboratory collection |
| JK093 | MO6-24/O with ΔiscR | 26 |
| JK128 | MO6-24/O with iscR 3CA encoding the apo-locked IscR, IscR3CA | 32 |
| HS03 | MO6-24/O with smcR::nptI; Kmr | 22 |
| DI0201 | MO6-24/O with Δcrp | 41 |
| SO111 | MO6-24/O with ΔgbpA | This study |
| KK141 | MORSR ΔgbpA, Rifr, Smr | This study |
| KK142 | MO6-24/O with ΔiscRΔcrp | This study |
| E. coli | ||
| S17–1λpir | λ-pir lysogen; thi pro hsdR hsdM+ recA RP4–2 Tc::Mu-Km::Tn7;Tpr Smr; host for π-requiring plasmids; conjugal donor | 29 |
| BL21 (DE3) | F− ompT hsdSB (rB− mB−) gal dcm (DE3) | Laboratory collection |
| Plasmids | ||
| pGEM-T Easy | PCR product cloning vector; Apr | Promega |
| pKK1401 | pGEM-T Easy with 390-bp fragment of gbpA upstream region; Apr | This study |
| pDM4 | R6K γ ori sacB; suicide vector; oriT of RP4; Cmr | 28 |
| pSO1101 | pDM4 with ΔgbpA Cmr | This study |
| pBS0907 | pDM4 with Δcrp; Cmr | 31 |
| pJK1113 | pKS1101 with nptI; Apr, Kmr | 32 |
| pKK1402 | pJK1113 with gbpA; Apr, Kmr | This study |
| pKK1403 | pJK1113 with iscR; Apr, Kmr | This study |
| pKK1404 | pJK1113 with crp; Apr, Kmr | This study |
| pKK1405 | pJK1113 with smcR; Apr, Kmr | This study |
| pET22a(+) | His6 tag fusion expression vector; Apr | Novagen |
| pSO1201 | pET22a (+) with gbpA; Apr | This study |
| pJK0928 | pET28a (+) with iscR; Apr | 31 |
| pHK0201 | pRSET A with crp; Apr | 41 |
| pHS104 | pRSET C with SmcR; Apr | 22 |
| pBBR-lux | Broad host range vector with luxCDABE operon; Cmr | 44 |
| pKK1407 | pBBR_lux with 555-bp fragment of gbpA upstream region; Cmr | This study |
| pKK1408 | pBBR_lux with 451-bp fragment of gbpA upstream region; Cmr | This study |
| pKK1409 | pBBR_lux with 411-bp fragment of gbpA upstream region; Cmr | This study |
| pKK1410 | pBBR_lux with 341-bp fragment of gbpA upstream region; Cmr | This study |
a The following abbreviations are used: Rif, rifampicin-resistant; Smr, streptomycin-resistant; Kmr, kanamycin-resistant; Tpr, trimethoprim-resistant; Apr, ampicillin-resistant; Cmr, chloramphenicol-resistant.