TABLE 1

Strains, plasmids, and primers used in this study

Strain, plasmid, or primerDescription or sequenceaReference or source
Strains
    B. megaterium DSM319Wild-type strain9
    B. megaterium WH321Genomic xylR mutant32
    B. megaterium WH377Genomic xylT mutant31
    B. subtilis DSM402B. subtilis subsp. subtilis strain 168 carrying the pBC16 plasmid (DSM23521)DSMZb
Plasmids
    pSSBm85Promoter-optimized shuttle vector for xylose-inducible production of GFP; PxylA-gfp10
    pRBBm99Derivative of pSSBm85 lacking xylR and PxylA-gfpThis work
    pKMMBm1XmaI site introduced at the 5′ end of xylR, and xylR stop codon replaced with a KpnI site, in pSSBm85This work
    pKMMBm2mCherry cloned into KpnI and AflII sites of pKMMBm1; xylR-mCherry fusionThis work
    pKMMBm5pSSBm85 ΔxylR mutantThis work
    pJS72pETDuet vector (Novagen) containing mCherry14
    pBC16Natural plasmid isolate originating from Bacillus cereus, precursor for pSSBm8526
Primers
    xylR-mCherry_for5′-TATCAGGTACCGTGAGCAAGGGCGAGGAG-3′This work
    xylR-mCherry_rev5′-TATCACTTAAGTTACTTGTACAGCTCGTCCATG-3′This work
    xylT_for5′-CGGAGGGTTACTGTTTGGATATGAC-3′This work
    xylT_rev5′-CCGTGTGCTAAAGATCCTAACCCTA-3′This work
    xylR_for5′-GCCTTGTAGATCGTCATCAGCAAA-3′This work
    xylR_rev5′-CGTATGCTCCTGCATTAGCTTCAT-3′This work
    gfp_for5′-TCTCGGACACAAACTCGAGTACAAC-3′This work
    gfp_rev5′-CTGCTAGTTGAACGGATCCATCTTC-3′This work
    rpoB_for5′-GGCGACGAAGTAGTAAAAGGTGAGA-3′This work
    rpoB_rev5′-GGCATCCTCATAGTTGTAACCATCC-3′This work
    gyrB_for5′-TACATGGTGTAGGTGCCTCAGTTGT-3′This work
    gyrB_rev5′-ACTTTTAAGTCAGCAGCCGGTACAC-3′This work
    heli1_for5′-GATGTAATCCATACGTCAGCTGTGC-3′This work
    heli1_rev5′-CCGGCTTCCTTGATTTATAACTGG-3′This work
    heli2_for5′-GTCGAACCTGTACACGGTGACTTTT-3′This work
    heli2_rev5′-GCTTCCGTAATAGGTCTGTTCATGC-3′This work
  • a In primer sequences, specific restriction enzymes used for cloning are underlined.

  • b DSMZ, German Collection of Microorganisms and Cell Cultures.