Table 1.

Bacterial strains and plasmids used or constructed in this study

Strain or plasmidRelevant propertiesaSource or reference
Strains
    Bacillus subtilis strains
        168 trpC2 35
        MGB625trpC2 Δprophage 1-6 ΔPBSX ΔSPβ Δpks Δskin Δpps Δ(ydeK-ydhU) Δ(yisB-yitD) Δ(yunA-yurT) Δ(cgeE-yodU) Δ(ypqP-ypmQ) Δ(yeeK-yesX) 39
        MGB723MGB625 Δ(pdp-rocR) 39
        MGB874MGB723 Δ(ycxB-sipU) Δ(yrkS-yraK) Δ(sboA-ywhH) Δ(yybP-yyaJ) Δ(yncM-yndN) 39
        MGB625 Δr1MGB625 Δ(pdp-yxeD)::catThis study
        MGB625 Δr2MGB625 Δ(yxeC-yxnA)::catThis study
        MGB625 Δr3MGB625 Δ(yxaD-fbp)::catThis study
        MGB625 Δr4MGB625 Δ(yydD-rocR)::catThis study
        MGB625 ΔyydD-yycNMGB625 Δ(yydD-yycN)::catThis study
        MGB625 ΔrapG-phrGMGB625 Δ(rapG-phrG)::catThis study
        MGB625 ΔrocF-rocRMGB625 Δ(rocF-rocR)::catThis study
        MGB625 ΔrocF-rocDMGB625 Δ(rocF-rocD)::catThis study
        MGB625 ΔrocRMGB625 ΔrocR::catThis study
        MGB625 Δpdp-phrGMGB625 Δ(pdp-phrG)::catThis study
        874DEFRMGB874 with cat-rocDEF-rocR inserted at the Δ(pdp-rocR) locusThis study
        874DEFR(spec)MGB874 with spec-rocDEF-rocR inserted at the Δ(pdp-rocR) locusThis study
        168 ΔgudBtrpC2 ΔgudB::neoThis study
        MGB874 ΔgudBMGB874 ΔgudB::neoThis study
        874DEFR ΔgudB874DEFR(cat) ΔgudB::neoThis study
        MGB874 ΔahrCMGB874 ΔahrC::neo (promoterless, terminatorless)This study
        MGB874 ΔargC-argFMGB874 ΔargCJBD-carAB-argF::specThis study
        MGB874 ΔahrC ΔargC-argFMGB874 ΔahrC ΔargCJBD-carAB-argF::specThis study
        MGB874 ΔcitBMGB874 ΔcitB::specThis study
        874DEFR ΔcitB874DEFR ΔcitB::specThis study
        MGB874 ΔrocGMGB874 ΔrocG::specThis study
        MGB874 ΔrocG(cat)MGB874 ΔrocG::catThis study
        MGB874 ΔgltABMGB874 ΔgltAB::ermThis study
        874DEFR ΔrocG874DEFR ΔrocG::specThis study
        874DEFR ΔgltAB874DEFR ΔgltAB::ermThis study
        168egl-237trpC2 ΔamyE::egl-237 catThis study
        MGB874egl-237MGB874 ΔamyE::egl-237 catThis study
        874DEFRegl-237874DEFR(spec) ΔamyE::egl-237 catThis study
    Escherichia coli strain
        HB101supE44 Δ(mcrC-mrr) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 leuB6 thi-1Takara Bio
Plasmids
    pHY300PLKShuttle vector for E. coli and B. subtilisTakara Bio
    pHYS237pHY300PLK carrying the gene for alkaline endo-1,4-β-glucanase (Egl-237) from Bacillus sp. strain KSM-S237, containing amp and tet 39
    pHP237-K16pHY300PLK carrying the gene for M protease from Bacillus clausii KSM-K16 fused to the promoter, SD, and leader peptide sequences of egl-237, containing amp and tet 39
    pXIntegration vector containing the Px promoter used for integration into the amyE locus by double crossover using cat 34
  • a Antibiotic resistance genes are shown as follows: cat, chloramphenicol; spec, spectinomycin; neo, neomycin; erm, erythromycin; amp, ampicillin; tet, tetracycline.