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Lysogenic bacteriophage M1 from Selenomonas ruminantium: isolation, characterization and DNA sequence analysis of the
Judy P E Cheong1, John D Brooker1
1Department of Animal Science, University of AdelaideWaite Campus, Glen Osmond 5064Australia.
Abstract:
Bacteriophage M1 from the ruminal bacterium Selenomonas ruminantium strain ML12 comprises a 30 nm icosahedral capsid, a 25 nm tail and 48 kb of linear dsDNA with cohesive ends. A restriction map of the phage genome has been constructed. The presence of bacteriophage M1 in the rumen has been demonstrated by PCR amplification and Southern blot analysis of DNA from rumen bacterial samples obtained from ten different sheep. Lysogeny was demonstrated by hybridization of M1 DNA to host chromosomal DNA and by identification and cloning of a 2.3 kb region of the phage containing the predicted attP domain which promotes chromosomal integration. DNA sequencing of the attP region demonstrated two major ORFs surrounding the predicted attP site and structural analysis of this region revealed a motif comprising three different inverted repeats surrounding a 12 bp palindrome. Analysis of the translated amino acid sequence upstream of the attP site demonstrated the presence of conserved residues found within integrase proteins of several temperate phages of different bacterial species.
Insights
Researchers characterized bacteriophage M1 from Selenomonas ruminantium, finding it integrates into the host genome. This study details the phage
Area of Science:
- Microbiology
- Virology
- Molecular Biology
Background:
- Bacteriophages are viruses that infect bacteria and play crucial roles in microbial ecosystems.
- Rumen bacteria, such as Selenomonas ruminantium, are key to host digestion.
- Understanding bacteriophage-host interactions is vital for microbial community dynamics.
Purpose of the Study:
- To characterize bacteriophage M1 isolated from Selenomonas ruminantium strain ML12.
- To investigate the integration mechanism and genetic elements involved in lysogeny.
- To confirm the presence of bacteriophage M1 in sheep rumen environments.
Main Methods:
- Isolation and characterization of bacteriophage M1.
- Genome analysis including restriction mapping and DNA sequencing of the attP region.
- Detection of phage presence in sheep rumen using PCR amplification and Southern blot analysis.
- Cloning and analysis of the attP site and surrounding open reading frames (ORFs).
Main Results:
- Bacteriophage M1 possesses an icosahedral capsid, a tail, and 48 kb of linear dsDNA with cohesive ends.
- The presence of bacteriophage M1 was confirmed in the rumen of ten different sheep.
- Lysogeny was demonstrated, with identification and cloning of a 2.3 kb attP region.
- Sequence analysis revealed conserved integrase motifs within the attP region, suggesting a temperate phage nature.
Conclusions:
- Bacteriophage M1 is a temperate phage capable of integrating into the Selenomonas ruminantium genome.
- The characterized attP region and associated genetic elements are crucial for phage integration.
- The findings contribute to understanding phage-host dynamics within the complex rumen ecosystem.