Pulsed-field gel electrophoresis of SmaI digests of lactococcal genomic DNA, a novel method of strain identification

Abstract

The pulsed-field gel electrophoresis (PFGE) pattern of SmaI digests of 29 strains of Lactococcus lactis subsp. lactis and subsp. cremoris were determined. Unrelated strains yielded markedly different patterns of digestion products. Bacteriophage-resistant derivatives of four strains, generated by a method analogous to that used regularly in some cheese factories, yielded patterns that were identical or almost identical to that of the parent strain. It is proposed that a 16-h PFGE run with a pulse time increasing linearly from 1 to 20 s, which separates fragments between 50 and 240 kilobase pairs (kbp) and produces a pattern containing around 15 bands, can be used as a reliable prrocedure for strain identification in the lactococci. SmaI digests of 24 of the strains were analyzed by PFGE at three different pulse times to determine accurately the sizes of fragments bigger than 8 kbp. The sum of the sizes of all of the fragments in the digest of a strain provided an estimate of the genome size of the strain. For all the strains analyzed, this estimate was within the range of 2.0 to 2.7 Mbp, with no apparent difference between L. lactis subsp. lactis, L. lactis subsp. lactis biovar diacetylactis and L. lactis subsp. cremoris strains.