TABLE 1

Strains and plasmids used in this study

Strain or plasmidRelevant propertiesaSource or reference
Strains
    E. coli strains
        DH5αF′ endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δ(argF-lacZYA)U169 ϕ80dlacΔM15Promega
        CC118 (λ pir)Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 λ pir58
        S17-1 (λ pir)Tpr Smr recA thi pro hsdR17(rK mK+) RP4::2-Tc::Mu Km Tn7 λ pir59
        BL21(DE3)F ompT hsdSB(rB mB) gal dcm (DE3)Invitrogen
    V. cholerae strains
        FY_Vc_2O1 El Tor A1552, rugose variant, Rifr23
        FY_Vc_5Fy_Vc_2 ΔvpsT Rifr24
        FY_Vc_6Fy_Vc_2 ΔvpsR Rifr24
        FY_Vc_9543FY_Vc_2/pFY_3406 Rifr CmrThis study
        FY_Vc_9544FY_Vc_6/pFY_3406 Rifr CmrThis study
        FY_Vc_9545FY_Vc_2/pFY_3407 Rifr CmrThis study
        FY_Vc_9546FY_Vc_2/pFY_3408 Rifr CmrThis study
        FY_Vc_9547FY_Vc_2/pFY_3409 Rifr CmrThis study
        FY_Vc_9548FY_Vc_2/pFY_3410 Rifr CmrThis study
        FY_Vc_9549FY_Vc_2/pFY_3411 Rifr CmrThis study
        FY_Vc_9550FY_Vc_2/pFY_3412 Rifr CmrThis study
        FY_Vc_9551FY_Vc_2/pFY_3413 Rifr CmrThis study
        FY_Vc_9552FY_Vc_2/pFY_3414 Rifr CmrThis study
        FY_Vc_9553FY_Vc_2/pFY_3415 Rifr CmrThis study
        FY_Vc_9554FY_Vc_2/pFY_3416 Rifr CmrThis study
        FY_Vc_9912FY_Vc_2/pFY_3493 Rifr CmrThis study
        FY_Vc_9556Fy_Vc_2 Δhns RifrThis study
        FY_Vc_9557FY_Vc_9556/pFY_3406 Rifr CmrThis study
        FY_Vc_9558FY_Vc_9556/pFY_3407 Rifr CmrThis study
        FY_Vc_9559FY_Vc_9556/pFY_3408 Rifr CmrThis study
        FY_Vc_9560FY_Vc_9556/pFY_3409 Rifr CmrThis study
        FY_Vc_9561FY_Vc_9556/pFY_3410 Rifr CmrThis study
        FY_Vc_9913FY_Vc_9556/pFY_3416 Rifr CmrThis study
        FY_Vc_9914FY_Vc_5/pFY_3406 Rifr CmrThis study
        FY_Vc_9915FY_Vc_5 Δhns-FRT-Kanr-FRT RifrThis study
        FY_Vc_9916FY_Vc_9915/pFY_3406 Rifr Kanr CmrThis study
Plasmids
    pMAL-c5xIPTG-inducible expression vector with N-terminal MBP, AprNEB
    pFY_1471pMAL-c5x-vpsT AprThis study
    pFY_1472pMAL-c5x-vpsR AprThis study
    pBBRluxluxCDABE-based promoter fusion vector, Cmr60
    pFY_3406pBBRlux vpsL promoter, Cmr (−607, +158)This study
    pFY_3407pBBRlux vpsL promoter with mutated R1 box, Cmr (−607, +158)This study
    pFY_3408pBBRlux vpsL promoter with mutated T box, Cmr (−607, +158)This study
    pFY_3409pBBRlux vpsL promoter with mutated R2a box, Cmr (−607, +158)This study
    pFY_3410pBBRlux vpsL promoter with mutated R2b box, Cmr (−607, +158)This study
    pFY_3411pBBRlux vpsL promoter, Cmr (−432, +158)This study
    pFY_3412pBBRlux vpsL promoter, Cmr (−379, +158)This study
    pFY_3413pBBRlux vpsL promoter, Cmr (−300, +158)This study
    pFY_3414pBBRlux vpsL promoter, Cmr (−272, +158)This study
    pFY_3415pBBRlux vpsL promoter, Cmr (−351, +158)This study
    pFY_3416pBBRlux vpsL promoter, Cmr (−250, +158)This study
    pFY_3493pBBRlux vpsL promoter, Cmr (−96, +158)This study
    pGP704-sacB28pGP704 derivative, mob-oriT sacB AprG. Schoolnik
    pFY_3418pGP704-sacB28-Δhns AprThis study
    pBR-FRT-Kanr-FRTpBR322 backbone61
  • a Numbers in parentheses indicate the nucleotide positions of the cloned vpsL regulatory regions with respect to the translation start site.