Table 1.

Strains, plasmids, and primers used in this work

Strain, plasmid, or primerDescriptionSource or reference
Strains
    Escherichia coli Mach1F φ80 (lacZ) ΔM15 ΔlacX74 hsdR(rK mK+) ΔrecA1398 endA1 tonATransgene
    Bacillus subtilis 168Type strainBacillus Genetic Stock Center (BGSC)
    B. subtilis WB600trpC2 ΔnprE ΔaprE Δepr Δbpf Δmpr ΔnprBBacillus Genetic Stock Center (BGSC)
    E. coli K-12Type strainLaboratory stock
Plasmids
    pEASY-T3Cloning vectorTransgene
    pEASY-T3-cotG-nanApEASY-T3 with fusion gene cotG-nanAThis study
    pHP13Low-copy-number E. coli-B. subtilis shuttle vectorAlan D. Grossman
    pHP13-cotG-nanApHP13 with fusion gene cotG-nanAThis study
    pDG1728E. coli-B. subtilis shuttle vector with spectinomycin resistance gene7
    pGJ103High-copy-number E. coli-B. subtilis shuttle vector39
    pEB03High-copy-number E. coli-B. subtilis shuttle vectorThis study
    pEB03-cotG-nanApEB03 with fusion gene cotG-nanAThis study
Primersa
    P15′-GCCTTTGAATTCAGTGTCCCTAGCTCCGAG-3′This study
    P2b5′-CTATTGACTAGTTGAACCCCCACCTCCTTTGTATTTCTTTTTGACTA-3′This study
    P35′-AGACTAGTATGGCAACGAATTTAC-3′This study
    P45′-TCCTGCAGTCACCCGCGCTCTT-3′This study
    Spc-up5′-TTGGGCCCGAATGGCGATTTTCGTTC-3′This study
    Spc-down5′-TGCGCTCGAGCCAATTAGAATGAATATTTC-3′This study
  • a The italicized letters indicate the introduction of restriction sites.

  • b The underlined letters indicate the introduction of a flexible linker at the C terminus of the cotG structural gene product.