TABLE 3

Strains and plasmids used in this study

Strain or plasmidGenotype or descriptionaReference, source, or transformationb
Strains
    B. subtilis
        YB955hisC952 metB5 leuC427 xin-1 Spβs11
        HB2078CU1065 ΔperR::kan Kanr46
        PERM1017ΔnrdE::lacZ Pspac-nrdEF::ery EryrpPERM1011 → YB955
        PERM1202ΔytcG::neo NeorpPERM1179 → YB955
        PERM1207ΔytcG::neo ΔnrdE::lacZ Pspac-nrdEF::ery Neor EryrpPERM1179 → PERM1017
        PERM1159ΔperR::kan ΔnrdE::lacZ Pspac-nrdEF::ery Kanr EryrpPERM1011 → HB2078
        PERM1448ytcG::lacZpPERM1330 → YB955
    E. coli XL10-GoldendA1 glnV44 recA1 thi-1 gyrA96 relA1 lac Hte Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)Tn10 Tetr CmrLaboratory stock
Plasmids
    pMUTIN4Integrational lacZ fusion vector; Ampr Eryr62
    pBEST501Integrational neomycin replacement vector; Ampr Neor63
    pPERM1011pMUTIN4 containing a 5′ fragment (from bp −47 to 290) of the nrdE ORF cloned into the HindIII-SacII sites; Ampr EryrThis study
    pPERM1179pBEST501 containing a 5′ fragment (from bp −367 to −22) and a 3′ fragment (from bp +23 to +365) adjacent to the ytcG ORF cloned into the HindIII-SalI and BamHI-SacI sites, respectively; Ampr EryrThis study
    pPERM1330pMUTIN4 containing a 3′ fragment (from bp −179 to 456) of the ytcG ORF cloned into BamHI-EcoRI sites; Ampr EryrThis study
  • a Amp, ampicillin; Ery, erythromycin; Neo, neomycin; Kan, kanamycin; ORF, open reading frame.

  • b Transformations are represented by X → Y, where strain Y was transformed with plasmid DNA from source X.