TABLE 1.

Bacterial strains, plasmids, and oligonucleotides used in this study

Strain, plasmid, or oligonucleotideDescription or sequenceAntibiotic resistanceReference or source
L. lactis subsp. cremoris strains
    MG1363Plasmid-free derivative of NCDO7126
    MP401MG1363 containing pMAP50Ermr23
    AJ189MG1363 containing pAJ149 (PL-lacLM)Ermr13
    AJ218MG1363 containing pAJ155 (PR-lacLM)Ermr13
Escherichia coli strains
    DH5αsupE44 hsdR17 deoR recA1 endA1 lacZΔM15Stratagene
    M15[pREP4]Expression host for pQE vectors, contains pREP4KanrQiagen
Plasmids
    pCI372Shuttle vectorCamr8
    pAJ149pAK80::P3 (3142-3458)a PL-lacLM fusion, containing OR and OLErmr13
    pAJ155pAK80::P3 (3142-3458)a PR-lacLM fusion, containing OR and OLErmr13
    pAJ80pCI372::PRORcI (2622-3332)Camr16
    pMAP50pAK80::cI+OR+OL+mor+OD+, PL-lacLMErmr23
    pMAP131pMAP50;15 base pairs inserted at codon 120 in cIErmrThis study
    pMAP132pMAP50;15 base pairs inserted at codon 98 in cIErmrThis study
    pMAP133pMAP50;15 base pairs inserted at codon 135 in cIErmrThis study
    pMAP140pMAP50;15 base pairs inserted at codon 78 in cIErmrThis study
    pMAP141pMAP50;15 base pairs inserted at codon 89 in cIErmrThis study
    pMAP150pCI372::PRcI; 15 base pairs inserted at codon 78CamrThis study
    pMAP151pCI372::PRcI; 15 base pairs inserted at codon 89CamrThis study
Oligonucleotides
    MLN6.rev5′-GGGGGGGTTTAAACATTTTAATTGAATTTTTGCAGTATC-3′
    morBamHI.rev5′-GGGGGATCCATGAGTTATGATTATTCGTCAC-3′
    MLN6.for5′-GGGGGGGTTTAAACATAAAAGAACAAGATGAGCAAAATAAAGTTA-3′
    pAK80erm5′-CTTTCAACTGCCTGGCAC-3′
    MLN8.rev5′-GGGGGGGTTTAAACAAACTGTAATAGTTTCTTCTATATT-3′
    MLN8.for5′-GGGGGGGTTTAAACACAGTTATGAAAAAATTGGAAGAGCCAAGGC-3′
    MLN27.rev5′-GGGGGGGTTTAAACATCAGACGGTAATCTTCTATTTGTT-3′
    MLN27.for5′-GGGGGGGTTTAAACATCTGAGTGATGAATATCTTGAAGAACAAAT-3
    MLN2.rev5′-GGGGGGGTTTAAACATACATCAAAGCCCATTAACCACGC-3′
    MLN2.for5′-GGGGGGGTTTAAACACAGTTCCTATGGTAGAATCTTCTAAAATA-3
    MLN3.rev5′-GGGGGGGTTTAAACAATCATTTTCTATTTTAGAAGATTC-3′
    MLN3.for5′-GGGGGGGTTRAAACACAGTTTCTGAAAATATAGAAGAAACTATT-3
    CIBamHI5′-GGGGGGGATCCGCTCTCTAAGTTTACTCATAAATCG-3
    erm2BamH5′-CGGGATCCCGCACGTTTCATGAACTTT-3′
  • a Numbers indicate nucleotide positions in the TP901-1 genome.