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Molecular characterization of the lactococcal plasmid pCIS3: natural stacking of specificity subunits of a type I restriction/modification system in a single lactococcal strain

National Food Biotechnology Centre¹,Department of Microbiology², and Department of Food Science and Technology³, University College Cork, Cork, Ireland

Author for correspondence: Gerald F. Fitzgerald. Tel: +353 21 902730. Fax: +353 21 903101. e-mail: g.fitzgerald{at}ucc.ie

A 6·1 kb plasmid from the Lactococcus lactis subsp. cremoris strain UC509.9, named pCIS3, was found to mediate a restriction/modification (R/M) phenotype. Nucleotide sequence analysis of pCIS3 revealed the presence of an hsdS gene, typical of type I R/M systems. The presence of this plasmid resulted in a 10⁴-fold reduction in the efficiency of plating (e.o.p.) of unmodified phage. In addition to the hsdS gene of pCIS3, two more hsdS genes were identified in strain UC509.9, one located on the chromosome downstream of a gene highly homologous to hsdM genes and a third on the smallest (4 kb) plasmid, named pCIS1. The replication region of pCIS3 was highly similar to that of a large family of lactococcal theta replicons. In addition, pCIS3 was found to encode a member of the CorA family of magnesium transporters.

Keywords: Lactococcus lactis, restriction, modification, magnesium transport, cobalt resistance

Abbreviations: abi, abortive infection; R/M, restriction/modification

The GenBank accession numbers for the sequences reported in this paper are AF153410–AF153414.

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