TABLE 1

Bacterial strains and plasmids used in this study

Strain or plasmidGenotype or phenotypeSource or reference
E. coli strains
    CGSC 8019F ΔmraZ52 ΔlacL Δfnr267 rph-140
    CGSC 8021F ΔmraW54 ΔlacL Δfnr267 rph-140
    DH5αF ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK mK+) supE44 λ thi-1 gyrA relA176
    DH5α pheF ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK mK+) supE44 λ thi-1 gyrA relA1 phe::Tn10dCm77
    JE6478MG1655/pJE6448 AprThis study
    JE6430MG1655/pDSW208 AprThis study
    JE6513MG1655/pJE6496 AprThis study
    JE6517MG1655/pJE6500 AprThis study
    JE6583MG1655/pJE6584 CmrThis study
    JE6598WM2909/pRK415 TetrThis study
    JE6599WM2909/pJE6595 TetrThis study
    JE6627MG1655 lacU169/pDSW208/pJE6618 Apr TetrThis study
    JE6628MG1655 lacU169/pJE6500/pJE6618 Apr TetrThis study
    JE6647MG1655 lacU169/pDSW208/pJE6621 Apr TetrThis study
    JE6648MG1655 lacU169/pJE6500/pJE6621 Apr TetrThis study
    JE6661MG1655/pJE6653 AprThis study
    JE6707MG1655 lacU169/pDSW208/pJE6695 Apr TetrThis study
    JE6708MG1655 lacU169/pJE6500/pJE6695 Apr TetrThis study
    JE6710MG1655 lacU169/pDSW208/pJE6696 Apr TetrThis study
    JE6711MG1655 lacU169/pJE6500/pJE6696 Apr TetrThis study
    JE6713MG1655 lacU169/pDSW208/pJE6697 Apr TetrThis study
    JE6714MG1655 lacU169/pJE6500/pJE6697 Apr TetrThis study
    JE7023MG1655 ΔmraZ52 MG1655 leuO::tet × P1(CGSC8019) Tets prototrophThis study
    JE7025MG1655 ΔmraW54 MG1655 leuO::tet × P1(CGSC8021) Tets prototrophThis study
    JE7031MG1655 ΔmraZ52/pDSW208This study
    JE7032MG1655 ΔmraZ52/pJE6500This study
    JE7035MG1655 ΔmraW54/pDSW208This study
    JE7108MG1655 lacU169/pJE6640/pJE6618 Apr TetrThis study
    MG1655Sequenced λ and F derivative of K-1278
    MG1655 lacU169lac mutant derivative of MG1655Laboratory strain
    WM2724W3110 with native ftsZ + Ptrc-ftsZ-gfp at λ att site79
    WM2909W3110ΔftsL::kan with Ts plasmid pTSA29-ftsL Kanr AprLaboratory strain
Plasmids
    pBAD33pACYC184 vector with PBAD promoter; Cmr80
    pBlueScriptII (pBSII)Apr, with T3 and T7 promotersStratagene
    pDSW208Ptrc promoter vector; Apr81
    pJE6448pDSW208 EcoRI-HindIII with mraZ-gfp; AprThis study
    pJE6496pDSW208 EcoRI-HindIII with mraW-flag; AprThis study
    pJE6500pDSW208 EcoRI-HindIII with mraZ-gluglu; AprThis study
    pJE6584pBAD33 KpnI-HindIII with mraZ-gluglu; CmrThis study
    pJE6595pRK415 BamHI-KpnI with ∼2,700-bp PCR fragment encompassing 563 bp upstream of mraZ to downstream ftsLThis study
    pJE6617pBSII SmaI with same ∼2,700-bp PCR fragment as pJE6595 but with MraZR15A mutationThis study
    pJE6618WT mraZ::lacZ translational fusion vector; pUI523A::593-bp PCR fragment from mraZ (563 bp upstream and 30 bp within gene); TetrThis study
    pJE6621O1 mraZ::lacZ translational fusion in pJE6618 with TGGGG-to-ATCGG mutation in DR2; TetrThis study
    pJE6640pDSW208 EcoRI-HindIII with mraZR15A-gluglu; AprThis study
    pJE6695O2 mraZ::lacZ fusion in pJE6618 with TGGGA-to-AGGCA mutation in DR1 and TGGGG to ATCGG mutation in DR2; TetrThis study
    pJE6696O3 mraZ::lacZ fusion in pJE6618 with TGGGG-to-ATCGG mutation in DR2 and TGGGA to ATCGA mutation in DR3; TetrThis study
    pJE6697O4 mraZ::lacZ fusion in pJE6618 with TGGGA-to-AGGCA mutation in DR1, TGGGG-to-ATCGG mutation in DR2, and TGGGA-to-ATCGA mutation in DR3; TetrThis study
    pJE6653pDSW208 EcoRI-HindIII with mraZ-mraW; AprThis study
    pJE6761pDSW208 NcoI-HindIII with his-mraZ-gluglu; AprThis study
    pJE6762pDSW208 NcoI-HindIII with his-mraZR15A-gluglu; AprThis study
    pJE7145pBAD33 KpnI-HindIII with mioCThis study
    pRK415Low-copy-number IncP plasmid; Tetr82
    pUI523APromoterless lacZ translational fusion vector; Tetr38