TABLE 1

Strains and plasmids

Strain or plasmidDescriptionReference or source
Strains
    E. coli S17-1λpirF recA hsdR RP4-2 (Tc::Mu) (Km::Tn7) lysogenized with λpir23
    V. cholerae
        C7258Wild-type O1, El Tor biotype, OgawaPeru, 1991
        C7258ΔlacZC7258 lacZ deletion mutant48
        AJB50C7258 ΔrpoS52
        AJB51C7258 ΔhapR52
        AJB51ΔlacZAJB51 ΔlacZ deletion mutant48
        HX11C7258 ΔlacZ ΔrpoS::rpoS-lacZThis study
        HX12C7258 ΔlacZ ΔhapR ΔrpoS::rpoS-lacZThis study
        HX13C7258 ΔvpsRThis study
        HX14C7258 ΔhapR ΔvpsRThis study
        HX15C7258 ΔlacZ ΔvpsR ΔrpoS::rpoS-lacZThis study
        HX16C7258 ΔlacZ ΔhapR ΔvpsR ΔrpoS::rpoS-lacZThis study
        HX17C7258 ΔvpsTThis study
        HX18C7258 ΔvpsR ΔvpsTThis study
        HX19C7258 ΔlacZ ΔvpsT ΔrpoS::rpoS-lacZThis study
        HX20C7258 ΔlacZ ΔhapR ΔvpsT ΔrpoS::rpoS-lacZThis study
        HX21C7258 ΔlacZ ΔvpsR ΔvpsT ΔrpoS::rpoS-lacZThis study
        HX03C7258 ΔhapR ΔvpsT22
        HX22C7258 vpsT::pCVDVpsT-FLAGThis study
        HX23C7258 ΔvpsR vpsT::pCVDVpsT-FLAGThis study
Plasmids
    pVIK111Suicide vector with promoterless lacZ; oriR6K Kmr27
    pCR-RpoS0.34-kb DNA fragment containing the rpoS promoter in pCR2.1This study
    pRpoS-Up0.8-kb SmaI-SalI DNA fragment encoding the rpoS promoter and the N-terminal 223 amino acids of RpoS in pUC19This study
    pRpoS-Down0.45-kb SphI-XbaI DNA fragment containing the 3′ terminus of the rpoS ORF in pUC19This study
    pLacZ3.2-kb SalI-SphI DNA fragment encoding the lacZ ORF from pVIK111 in pUC19This study
    pRpoS-Up-lacZ3.2-kb SalI-SphI DNA fragment from pLacZ in pRpoS-UpThis study
    pCVDRpoS-Up-lacZ4-kb SmaI-SphI DNA fragment from pRpoS-Up-lacZ in pCVD442This study
    pCVDΔRpoS-lacZ0.45-kb SphI-XbaI DNA fragment from pRpoS-Down in pCVDRpoS-Up-lacZThis study
    pUC-VpsR-UP0.5-kb XbaI-SalI DNA fragment containing the 5′ terminus of vpsR in pUC19This study
    pUC-VpsR-Down0.5-kb SalI-SphI DNA fragment containing the 3′ terminus of vpsR in pUC19This study
    pUCΔvpsRXbaI-SalI and SalI-SphI DNA fragments flanking the vpsR ORF sequentially cloned into pUC19This study
    pCVDΔvpsRXbaI-SphI fragment harboring a vpsR deletion in pCVD442This study
    pAT1662DGC VCA0956 cloned into pBAD3325
    pCR2.1Bacterial expression vector for TA cloning (Ampr)Invitrogen
    pCR-VpsT10.67-kb XhoI-KpnI DNA fragment encoding the vpsT ORF in pCR2.1This study
    pCR-VpsT20.67-kb NdeI-SapI PCR fragment encoding the vpsT ORF in pCR2.1This study
    pTXB1Expression vector for construction of in-frame fusions with the intein/chitin binding domainNew England BioLabs
    pTXB1-VpsTVector pTXB1 containing a VpsT-intein fusionThis study
    pFLAG-CTCVector for cytoplasmic expression of C-terminal FLAG fusionsSigma-Aldrich
    pFLAG-CTC*pFLAG-CTC with modified multicloning site lacking the SalI restriction siteThis study
    pTT3rrnB T1T2 transcription terminator in pUC1928
    pTT70.43-kb SalI-BamHI DNA fragment encoding the rrnB T1T2 transcription terminator in pUC19This study
    pTT7VpsT-FLAG0.73-kb BamHI-SmaI DNA fragment containing the vpsT ORF and FLAG epitope coding region in pTT7This study
    pCVDVpsT-FLAGSalI-SmaI vpsT-FLAG and rrnB T1T2 fragment in pCVD442This study
    pRpoSLac5An SphI-HindIII DNA fragment containing the rpoS promoter region cloned between the rrnB T1T2 transcription terminator and a promoterless lacZ gene in pKRZ1a29
    pACTluz-RpoSrrnB T1T2-rpoS promoter fragment from pRpoSLac5, promoterless luxCDABE operon, and OriT in pACYC184This study
    pCVDΔvpsT-KmpCVD442-based vector with vpsT deletion and Km insertion22
    pCMW75V. harveyi IPTG-inducible DGC QrgB expression vector20
  • a See reference 53.