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Prophage Curing and Partial Characterization of Temperate Bacteriophages from Thermolytic Strains of Lactococcus lactis ssp. cremoris

Department of Food Science, University of Wisconsin, Madison 53706

The thermolytic response observed in two strains (SK11 and US3) of Lactococcus lactis ssp. cremoris was investigated. Lysis was induced 2 to 2.5 h after either strain was transferred from 30 to 37°C or exposed to mitomycin C. The temperature shift resulted in slow, incomplete lysis. Temperate phages were isolated following mitomycin C induction from both strains SK11 (SK11-T1) and US3 (US3-T1). Restriction patterns of DNA isolated from these phages were indistinguishable upon agarose gel electrophoresis. In total genomic DNA preparations, phage DNA bands developed 1.5 to 2 h after mitomycin C treatment. No phage DNA bands were observed during lysis induced by the temperature shift. Analysis of concentrated phage samples by transmission electron microscopy showed a multitude of intact US3-T1 phage from mitomycin C induction of US3. In samples from temperature inductions of US3, only a few intact bacteriophage particles with a different appearance were observed. A prophage-cured derivative of strain US3, designated US3-12, was obtained after exposure to UV irradiation. US3-12 continued to lyse in the presence of both mitomycin C and increased temperature; however, its mitomycin C lysis curve then resembled its lysis curve induced by the temperature shift.

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