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DNA Sequence Analysis of Three Lactococcus lactis Plasmids Encoding Phage Resistance Mechanisms

  • I. Boucher
    Affiliations
    Department of Biochemistry and Microbiology, Faculté des Sciences et de Génie, Groupe de Recherche en Écologie Buccale (GREB), Faculté de Médecine Dentaire, Université Laval, Québec, Canada, G1K 7P4
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  • Author Footnotes
    1 Current address: Chr. Hansen, Inc. 9015 West Maple Street, Milwaukee, WI 53214-4298.
    É. Émond
    Footnotes
    1 Current address: Chr. Hansen, Inc. 9015 West Maple Street, Milwaukee, WI 53214-4298.
    Affiliations
    Department of Food Science and Nutrition, Centre de Recherche en Science et Technologie du Lait (STELA), Faculté des Sciences de l’Agriculture et de l’Alimentation, Université Laval, Québec, Canada, G1K 7P4
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  • M. Parrot
    Affiliations
    Department of Biochemistry and Microbiology, Faculté des Sciences et de Génie, Groupe de Recherche en Écologie Buccale (GREB), Faculté de Médecine Dentaire, Université Laval, Québec, Canada, G1K 7P4
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  • S. Moineau
    Correspondence
    Corresponding author.
    Affiliations
    Department of Biochemistry and Microbiology, Faculté des Sciences et de Génie, Groupe de Recherche en Écologie Buccale (GREB), Faculté de Médecine Dentaire, Université Laval, Québec, Canada, G1K 7P4
    Search for articles by this author
  • Author Footnotes
    1 Current address: Chr. Hansen, Inc. 9015 West Maple Street, Milwaukee, WI 53214-4298.
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      Abstract

      The three Lactococcus lactis plasmids pSRQ700, pSRQ800, and pSRQ900 encode the previously described anti-phage resistance mechanisms LlaDCHI, AbiK, and AbiQ, respectively. Since these plasmids are likely to be introduced into industrial Lactococcus lactis strains used to manufacture commercial fermented dairy products, their complete DNA sequences were determined and analyzed. The plasmids pSRQ700 (7784 bp), pSRQ800 (7858 bp), and pSRQ900 (10,836 bp) showed a similar genetic organization including a common lactococcal theta-type replicon. A second replication module showing features of the pMV158 family of rolling circle replicons was also found on pSRQ700. The theta replication regions of the three plasmids were associated with two additional coding regions, one of which encodes for HsdS, the specificity subunit of the type I restriction/modification system. When introduced into L. lactis IL1403, the HsdS of pSRQ800 and pSRQ900 conferred a weak resistance against phage P008 (936 species). These results indicated that both HsdS subunits can complement the chromosomally encoded type I restriction/modification system in IL1403. The genes involved in the phage resistance systems LlaDCHI, AbiK, and AbiQ were found in close proximity to and downstream of the replication modules. In pSRQ800 and pSRQ900, transfer origins and putative tyrosine recombinases were found upstream of the theta replicons. Genes encoding recombination proteins were also found on pSRQ700. Finally, open reading frames associated with bacteriocin production were found on pSRQ900, but no anti-lactococcal activity was detected. Based on our current knowledge, these three plasmids are safe and suitable for food-grade applications.

      Abbreviation key:

      EOP (efficiency of plaquing), IR (inverted repeats), IRR (right inverted repeat), IS (insertion sequence), ORF (open reading frame), RBS (ribosome binding site), RC (rolling circle), R/M (restriction/modification), SD (Shine-Dalgarno)

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