TABLE 1.

Strains and plasmids used in this study

Strain or plasmidRelevant genotype and phenotypeaReference or source
Strains
    S. aureus
        RN4220NCTC8325-4 r m+rsbU 21
        COL mec; high-Mcr clinical isolate; Mcr Tcr 20
        NewmanClinical isolate (ATCC 25904); CP-5 producer 9
        BF21RN6390 arlR::cat; phenotypic arlRS mutant; Cmr 10
        GP268NCTC8325 derivative; (rsbU-V-W-sigB)+-tet(L); Tcr 12
        IK181NCTC8325 derivative; ΔrsbUVW-sigB::erm(B); Ermr 22
        IK183COL ΔrsbUVW-sigB::erm(B); Ermr 22
        IK184Newman ΔrsbUVW-sigB::erm(B); Ermr 22
        SM1RN4220 ΔspoVG::erm(B); phenotypic yabJ-spoVG mutant, ErmrThis study
        SM2Newman ΔspoVG::erm(B); phenotypic yabJ-spoVG mutant, ErmrThis study
        SM99Newman arlR::cat; CmrThis study
    E. coli
        XL1Blue recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB laclqZΔM15 Tn10 (Tcr)]Stratagene
Plasmids
    pAC7Expression plasmid containing the PBAD promoter and the araC gene; Cmr 42
    pAC7-sigBpAC7 with a 0.75-kb PCR fragment of the sigB ORF from strain COL; Cmr 14
    pBT1.6-kb PCR fragment of the tet(L) gene of pHY300PLK into Alw26I-digested pBC SK(+) (Stratagene); Tcr 12
    pBus1 E. coli-S. aureus shuttle plasmid with multicloning site from pBluescript II SK (Stratagene) and the rrnT14 terminator sequence from pLL2443; Tcr 45
    pEC1pUC derivative containing the 1.45-kb ClaI erm(B) fragment of Tn551; Apr Emr 4
    pGC2 E. coli-S. aureus shuttle plasmid; CmrConstructed and obtained by P. Matthews
    pSA0455ppSB40N with a 360-bp PCR fragment covering the σB-dependent promoter region preceding yabJ-spoVG fused to the reporter gene lacZα; Apr 3
    pSB40NPromoter probe plasmid; Apr 19
    pSP-luc+Firefly luciferase cassette vector; AprPromega
    pSTM01pEC1 with 0.5- and 1.1-kb PCR fragments covering the spoVG flanking regions; Emr AprThis study
    pSTM02pBT with a 3.1-kb KpnI/HindIII fragment of pSTM01 harboring the spoVG flanking regions and the erm(B) cassette fully replacing the spoVG coding region; Emr TcrThis study
    pSTM03pSP-luc+ with a 0.4-kb PCR fragment covering the capA promoter region fused to the reporter gene luc+; Apr EmrThis study
    pSTM04pBus1 with a 2.1-kb KpnI/EcoRI fragment of pSTM03 harboring the capA promoter-luc+ fusion; TcrThis study
    pSTM05pGC2 with a 1-kb PCR fragment covering the σB-dependent yabJ promoter, yabJ, and spOVG; CmrThis study
    pSTM06pSB40N with a 0.8-kb PCR fragment covering the arlRS promoter fused to the reporter gene lacZα; AprThis study
    pSTM07pSB40N with a 0.4-kb PCR fragment covering the capA promoter region fused to the reporter gene lacZα; AprThis study
    pSTM11pBus1 with a 0.6-kb PCR fragment covering yabJ and the preceding σB-dependent promoter PyabJ; TcrThis study
    pSTM13pBus1 with a 0.5-kb PCR hybrid including the σB-dependent promoter PyabJ fused to the spoVG ORF; TcrThis study
  • a Abbreviations are as follows: Apr, ampicillin resistant; Cmr, chloramphenicol resistant; Emr, erythromycin resistant; Tcr, tetracycline resistant.