TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidGenotype or descriptionSource or reference
B. subtilis 168 strains
    KSS1001trpC2K. Bott
    KSS1570amyE::pdivIVB mreD-minCDThis study
    KSS1571Ω pLEE100, amyE::pdivIVB mreD-minCDThis study
    KSS1572trpC2, Ω pLEE100This study
    KSS1573KSS1571 (pLEE504)This study
    MO1099amyE::erm9
E. coli strains
    DH5αendA1 gyrA96 hsdR17 ΔlacU169 (φ80lacZΔM15) recA1 supE44 thi-1BRLc
    M15Strr lacZΔM15 (pRep4 Kmr)Qiagen
Plasmids
    pCHA2489mreC ORFa PCR product cloned into pT7BlueThis study
    pCHA2506NotI (in mreC, blunted)-SalI (vector site) mreC DNA fragment from pCHA2489 cloned into EcoICRI-SalI site of pQE30This study
    pCR2.1PCR fragment cloning vector, Apr KmrInvitrogen
    pDG364amyE locus integration vectorBGSCd (9)
    pDG3053Shuttle expression plasmid utilizing the S. aureus lactose operon promoter; Apr KanrThis study
    pLEE50pUS19 with the lacI spac EcoRI-BamHI insert from pSI-1This study
    pLEE100MreC-inducible expression integration vectorThis study
    pLEE200pDG364 containing pdivIVB mreC-minCDThis study
    pLEE504pDG3053 containing truncated mreCbThis study
    pQE30Six-histidine tag vector, AprQiagen
    pSI-1IPTG-inducible expression shuttle plasmidD. Henner (2, 33)
    pT7BluePCR product cloning vector, AprInvitrogen
    pUS19pUC19 with Spr expressed in B. subtilis3
  • a ORF, open reading frame.

  • b Truncated mreC (contains bp 1 to 540).

  • c BRL, Bethesda Research Laboratories.

  • d BGSC, Bacillus Genetic Stock Center, Ohio State University.