TABLE 2

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant characteristicsReference or source
E. coli strains
    DH10βHost for cloning and vector constructionLaboratory stock
    JM110dam dcm mutant, host for preparation of the unmethylated plasmidLaboratory stock
S. aureus KCTC 3881Source of crtM-crtN genesKCTCa
B. thuringiensis strainsb
    BMB171Parental strain for all strains constructed in this studyBGSCc
    YBT-020Source of the slpA promoterBGSC
    BT-001BMB171 BMB171_C4312::crtM-crtNThis study
    BT-003BMB171 ΔsspAThis study
    BT-004BMB171 ΔsspBThis study
    BT-005BMB171 ΔsspA ΔsspBThis study
    BT-006BMB171 spo0A::loxPdThis study
    BT-007BMB171 Δspo0AThis study
    BT-008BMB171 spo0A::loxP ΔBMB171_C4815::PC0536-creeThis study
    BT-009BMB171 spo0A::loxP ΔBMB171_C1559::PC0536-creThis study
    BT-010BMB171 spo0A::loxP ΔBMB171_C4815::PC4286-crefThis study
    BT-011BMB171 spo0A::loxP ΔBMB171_C1559::PC4286-creThis study
    BT-016BMB171 BMB171_C4312::crtM-crtN spo0A::loxP ΔBMB171_C4815::PC0536-cre ΔplcR ΔsspA ΔsspBThis study
Plasmids
    pUC19Backbone vector for pAD01NEBg
    pDG1662Source of spectinomycin markerBGSC
    pUCTV2Source of tetracycline marker and Tsh replicon25
    pJW168Source of Cre recombinase geneLucigeni
    pAD01Ampr is replaced by Spcr; I-SceI recognition site was introducedThis study
    pAD02I-SceI expression vector with slpA promoter; Ts repliconThis study
    pAD03FLP expression vector with slpA promoter; Ts repliconThis study
    pAD04plcR knockout vectorThis study
    pAD05sspA knockout vectorThis study
    pAD06Himar1 transposon delivery vector with crtM-crtN; Ts repliconThis study
    pAD07sspB knockout vectorThis study
    pAD08loxP insertion vector at the spo0A locusThis study
    pAD09spo0A knockout vectorThis study
    pAD10Cre recombinase expression cassette (driven by the BMB171_C0536 promoter) insertion vector at the BMB171_C4815 locusThis study
    pAD11Cre recombinase expression cassette (driven by the BMB171_C0536 promoter) insertion vector at the BMB171_C1559 locusThis study
    pAD12Cre recombinase expression cassette (driven by the BMB171_C4286 promoter) insertion vector at the BMB171_C4815 locusThis study
    pAD13Cre recombinase expression cassette (driven by the BMB171_C4286 promoter) insertion vector at the BMB171_C1559 locusThis study
  • a KCTC, Korean Collection for Type Cultures (Jeongeup, Republic of Korea).

  • b Locus tag names were used instead of gene names when gene names were not available (e.g., for hypothetical proteins) or three-letter symbols were not available.

  • c BGSC, Bacillus Genetic Stock Center (Columbus, OH, USA).

  • d Two loxP sites were inserted at both ends of the spo0A gene.

  • e Cre recombinase expression cassette driven by the BMB171_C0536 promoter.

  • f Cre recombinase expression cassette driven by the BMB171_C4286 promoter.

  • g New England Biolabs (Ipswich, MA, USA).

  • h Ts, temperature sensitive.

  • i Lucigen Corporation (Middleton, WI, USA).