Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidCharacteristicsSource or reference
V. vulnificus
 ATCC 29307Type strainATCCa
 MO6-24/OClinical isolateJ. G. Morris, Jr.
 CMM981MO6-24/O withtoxR null mutationThis study
E. coli strains
 DH5αF recA1; restriction negativeLaboratory collection
 VM2SY327 [1NFVM1 F(ctx′-lacZ +)] 29
 SY327 λ pir D(lac pro)argE(Am) rif nalA recA56 λ pirlysogen 30
 SM10 λpir thi thr leu tonA lacY supE recA::RP4-2-Tcr:Mu Kmr λ pir lysogen 30
 INVαF′Competent cell line used for PCR TA cloningInvitrogen
 XL2 blue-MRF′Competent cell line used for genomic library constructionStratagene
Plasmids
 pCRIIPCR TA cloning vectorInvitrogen
 pBAD24PBAD expression vector 14
 pACYC184TcrCmr Laboratory collection
 pUC18, pUC19Apr α-lac Laboratory collection
 pGEX-2TGST fusion expression vectorPharmacia
 pGEX-3XGST fusion expression vectorPharmacia
 pNQ705Suicide vector with R6K origin; Cmr 32
 pLAFR3IncP broad-host-range plasmid; Tcr Laboratory collection
 pCVD7023.4-kb fragment containing vvh cloned intoHindIII-EcoRI sites of pBR325 55
 pJL9613.4-kb vvhfragment in pCVD702 cloned into SalI site of pACYC184This study
 pCMM9pCRII containing PCR-amplified 864-bp fragment carrying parts of toxR Vv andtoxS Vv This study
 pCMM12pCRII containing PCR-amplified 1,450-bp fragment carrying entiretoxRS Vv genesThis study
 pCMM51pUC19 containing 1,582-bpHindIII-BglII fragment carrying part oftoxR Vv and entire toxSgeneThis study
 pCMM74pUC19 containing 1,115-bpHindIII fragment carrying parts oftoxR Vv and htpG VvgenesThis study
 pCMM601 toxRS Vvcloned into EcoRI site of pBAD24This study
 pCMM610 toxR Vv cloned intoEcoRI site of pBAD24This study
 pCMM641 toxRS Vc cloned intoKpnI-EcoRI sites of pBAD24This study
 pCMM651 toxR Vc cloned intoKpnI-EcoRI sites of pBAD24This study
 pCMM700 KpnI-HindIII fragment of pCMM9 cloned into pNQ705This study
 pCMM709 toxR Vv cloned intoBamHI-EcoRI sites of pGEX-3XThis study
 pCMM820 toxS Vv cloned intoBamHI-EcoRI sites of pGEX-2TThis study
 pCMM20012.7-kbHindIII-HindIII-BglII fragment containing toxRS Vv operon and part ofhtpG Vv cloned into pLAFR3This study
  • a ATCC, American Type Culture Collection.