TABLE 1.

Strains and plasmids used in this study

Strain or plasmidCharacteristics
V. cholerae strains
    C7258Wild type, E1 Tor (Peru isolate, 1991)
    C6709-1Wild type, E1 Tor (Peru isolate, 1991)
    C6706Wild type, E1 Tor (Peru isolate, 1991)
    N16961E1 Tor, hapR (naturally occurring frameshift in hapR)
    AC-V66C6709-1, lacZ: res-tet-res (A. Camilli, Tufts University) (29)
    KSK394C6706, crp:: Kmr (K. Skorupski, Dartmouth Medical School) (46)
    DSM-V491C6709-1, ΔrpoS (A. Camilli, Tufts University) (32)
    AJB2AC-V66, hapA::hapA-lacZ, Ampr (this study)
    AJB3KSK394, hapA::hapA-lacZ, Ampr (this study)
    AJB19C7258 relA::pCVDΔrelAK (this study)
    AJB216N16961, ΔaphA (this study)
    AJB231AC-V66, ΔaphA (this study)
    AJB25DSM-V491, hapR::hapR-lacZ (this study)
    AJB26AC-V66, hapR::hapR-lacZ (this study)
    AJB29AC-V66, rpoS::rpoS-lacZ (this study)
E. coli strains
    DH5αF φ80lacZM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 (rK mR+) phoA supE44 λ thi-1 gyrA96 relA1
    SY327λPirΔ(lac pro) argE(Am) rif nalA recA56pirR6K) (35)
    SM10λPirthi thr leu tonA lacY supE recA::RP4-2Tc::Mu (λpirR6K) (35)
Plasmids
    pBADHisBExpression vector with araBAD promoter, Ampr (Invitrogen)
    pALTERex2In vitro mutagenesis vector, T7 promoter, Tetr (Promega Corp.)
    pGP704Suicide vector (oriR6K, mobRP4, Ampr) (35)
    pCVD442Suicide vector (oriR6K, mobRP4, sacB, Ampr) (11)
    pHapLac11Plasmid vector containing transcriptional hapA-lacZ fusions (45)
    pHapRLac2Plasmid vector containing transcriptional hapR-lacZ fusions (45)
    pGPHap10KpnI-PstI hapA-lacZ fusion from pHaplac11 in pGP704 (this study)
    pGPHap13KpnI-PstI hapR-lacZ fusion from pHapRlac2 in pGP704 (this study)
    pCH2pACYC184 containing 3.2-kb HindIII hapA insert (19)
    pBADHapR9HapR ORF cloned in pBADHisB (this study)
    pBADCRP7CRP ORF cloned in pBADHisB (this study)
    pCVDΔrelA1,077-bp internal relA fragment cloned in pCVD442 (this study)
    pCVDΔrelAKpCVDΔrelA with Kmr in unique NsiI site of relA (this study)
    pΔAphA5′ SphI-BamHI and 3′ BamHI-SacI aphA fragments in pUC19 (this study)
    pCVDΔAphASphI-SacI ΔAphA fragment from pΔAphA cloned in pCVD442 (this study)
    pRpoS5SphI-HindIII rpoS promoter fragment cloned in pUC19 (this study)
    pRpoSlac5SphI-HindIII rpoS DNA replacing hapR promoter in pHapRlac2 (this study)
    pGPRpoSlacZXbaI-ScaI fragment from pRpoSlac5 cloned in pGP704 (this study)
    pJB43428-bp hapA 5′ fragment in pALTERex2 (this study)