TABLE 1

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionReference or source
Bacterial strains
    E. faecalis strains
        V583Bloodstream isolate; VanB-type VRE13
        OG1RFHuman oral isolate33
        ΔchlRE. faecalis V583 ΔchlRThis study
        ΔefrEFE. faecalis V583 ΔefrEFThis study
        ΔefrEF/pFL103E. faecalis V583 ΔefrEF transformed with pFL103This study
        FL101E. faecalis V583 transformed with pPB101This study
        FL201E. faecalis V583 transformed with pFL201This study
        FL202E. faecalis V583 transformed with pFL202This study
        FL203E. faecalis V583 transformed with pFL203This study
        FL204E. faecalis V583 transformed with pFL204This study
        ΔchlR/pFL102ΔchlR mutant transformed with pFL102This study
        ΔchlR/pFL201ΔchlR mutant transformed with pFL201This study
        ΔchlR/pFL202ΔchlR mutant transformed with pFL202This study
    E. coli strains
        EC1000Cloning host; provides repA in trans; F araD139 (ara ABC-leu)7679 galU galK lacX74 rspL thi repA of pWV01 in glgB Km34
        DH5αCloning host; F endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 hsdR17(rK mK+) λ35
        BW23474Cloning host for pPB101 and derivatives; Δlac-169 robA1 cre C510 hsdR514 endA recA1 ΔuidA::pir-11636
Plasmids
    pHA101pLT06 plasmid with oriT from pHOU2 inserted at PstI21
    pCAT28Shuttle vector; pUC and pAMβ1 origins; confers chloramphenicol resistanceM. Rodrigues and K. Palmer
    pFL102pCAT28 containing 882-bp EcoRI/BamHI-digested chlR ORF and promoter regionThis study
    pFL103pCAT28 containing 3,673-bp EcoRI/BamHI-digested efrEF ORF and promoter regionThis study
    pPB101pTCV-lac-cat; expression vector for Gram-positive bacteria; confers kanamycin, erythromycin, and chloramphenicol resistance21
    pFL201pPB101 with 114-bp EcoRI/BamHI-digested efrEF promoter regionThis study
    pFL202pPB101 with 114-bp EcoRI/BamHI-digested chlR promoter regionThis study
    pFL203pPB101 with 98-bp EcoRI/BamHI-digested efrEF promoter regionThis study
    pFL204pPB101 with 98-bp EcoRI/BamHI-digested efrEF promoter region with modified motif CAGCTACThis study