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A temperate phage with cohesive ends induced by mitomycin C treatment of Lactobacillus casei
Y Nakashima1, H Hasuwa, Y Kakita
1Microbiology Laboratory, Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Abstract:
A temperate phage, named PL-2, was induced from Lactobacillus casei ATCC 27092 by mitomycin C treatment of the cells at exponential growth phase. The phage had an isometric head of 45 nm in diameter and a flexible, non-contractile tail, 150 nm long and 10 nm wide, with a sharp tip. Along the tail axis, about 40 regularly spaced striae were seen. The phage DNA had complementary cohesive ends. The restriction enzyme map of the DNA was constructed by using 13 different restriction endonucleases. The size of the DNA was 35.2 kb, 83% in size of that of phage PL-1 lytic for the same Lb. casei strain.
Insights
Researchers identified a new temperate phage, PL-2, from Lactobacillus casei. This phage has a distinct morphology and its DNA was mapped using restriction enzymes, revealing cohesive ends and a size of 35.2 kb.
Area of Science:
- Microbiology
- Virology
- Molecular Biology
Background:
- Lactobacillus casei is a significant probiotic bacterium.
- Temperate phages can influence bacterial host properties and are relevant in food fermentation and biotechnology.
- Characterization of novel phages is crucial for understanding phage-host interactions and potential applications.
Purpose of the Study:
- To isolate and characterize a novel temperate phage, designated PL-2, from Lactobacillus casei ATCC 27092.
- To determine the morphological and genomic features of phage PL-2.
- To construct a restriction enzyme map of the phage PL-2 genome.
Main Methods:
- Induction of temperate phage from Lactobacillus casei using mitomycin C.
- Transmission electron microscopy for phage morphology analysis.
- DNA extraction, restriction endonuclease digestion, and gel electrophoresis for genome analysis.
Main Results:
- A temperate phage, PL-2, was successfully induced from Lactobacillus casei.
- Phage PL-2 exhibits an isometric head (45 nm) and a flexible, non-contractile tail (150 nm x 10 nm) with visible striae.
- The phage DNA is 35.2 kb in size, possesses complementary cohesive ends, and its restriction map was elucidated using 13 enzymes.
Conclusions:
- Phage PL-2 is a novel temperate bacteriophage of Lactobacillus casei with unique structural and genomic characteristics.
- The detailed restriction map provides a foundation for further genetic studies of phage PL-2.
- The findings contribute to the understanding of phage diversity within Lactobacillus species.