TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Characteristics | Source or reference |
|---|---|---|
| V. vulnificus | ||
| ATCC 29307 | Type strain | ATCCa |
| MO6-24/O | Clinical isolate | J. G. Morris, Jr. |
| CMM981 | MO6-24/O with toxR null mutation | This study |
| E. coli strains | ||
| DH5α | F−recA1; restriction negative | Laboratory collection |
| VM2 | SY327 [1NFVM1 F(ctx′-lacZ+)] | 29 |
| SY327 λ pir | D(lac pro) argE(Am) rif nalA recA56 λ pir lysogen | 30 |
| SM10 λ pir | thi thr leu tonA lacY supE recA::RP4-2-Tcr:Mu Kmr λ pir lysogen | 30 |
| INVαF′ | Competent cell line used for PCR TA cloning | Invitrogen |
| XL2 blue-MRF′ | Competent cell line used for genomic library construction | Stratagene |
| Plasmids | ||
| pCRII | PCR TA cloning vector | Invitrogen |
| pBAD24 | PBAD expression vector | 14 |
| pACYC184 | Tcr Cmr | Laboratory collection |
| pUC18, pUC19 | Apr α-lac | Laboratory collection |
| pGEX-2T | GST fusion expression vector | Pharmacia |
| pGEX-3X | GST fusion expression vector | Pharmacia |
| pNQ705 | Suicide vector with R6K origin; Cmr | 32 |
| pLAFR3 | IncP broad-host-range plasmid; Tcr | Laboratory collection |
| pCVD702 | 3.4-kb fragment containing vvh cloned into HindIII-EcoRI sites of pBR325 | 55 |
| pJL961 | 3.4-kb vvh fragment in pCVD702 cloned into SalI site of pACYC184 | This study |
| pCMM9 | pCRII containing PCR-amplified 864-bp fragment carrying parts of toxRVv and toxSVv | This study |
| pCMM12 | pCRII containing PCR-amplified 1,450-bp fragment carrying entire toxRSVv genes | This study |
| pCMM51 | pUC19 containing 1,582-bp HindIII-BglII fragment carrying part of toxRVv and entire toxS gene | This study |
| pCMM74 | pUC19 containing 1,115-bp HindIII fragment carrying parts of toxRVv and htpGVv genes | This study |
| pCMM601 | toxRSVv cloned into EcoRI site of pBAD24 | This study |
| pCMM610 | toxRVv cloned into EcoRI site of pBAD24 | This study |
| pCMM641 | toxRSVc cloned into KpnI-EcoRI sites of pBAD24 | This study |
| pCMM651 | toxRVc cloned into KpnI-EcoRI sites of pBAD24 | This study |
| pCMM700 | KpnI-HindIII fragment of pCMM9 cloned into pNQ705 | This study |
| pCMM709 | toxRVv cloned into BamHI-EcoRI sites of pGEX-3X | This study |
| pCMM820 | toxSVv cloned into BamHI-EcoRI sites of pGEX-2T | This study |
| pCMM2001 | 2.7-kb HindIII-HindIII-BglII fragment containing toxRSVv operon and part of htpGVv cloned into pLAFR3 | This study |
a
ATCC, American Type Culture Collection.