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An origin of DNA replication from Lactococcus lactis bacteriophage c2
N R Waterfield1, M W Lubbers, K M Polzin
1Department of Pathology, University of Cambridge, England.
Applied and Environmental Microbiology
|April 1, 1996
Summary
Researchers identified a DNA replication origin (ori) in lactococcal phage c2, functional in Lactococcus lactis but not E. coli. This discovery enables new lactococcal and shuttle vector development.
Area of Science:
- Microbiology
- Molecular Biology
- Virology
Background:
- Bacteriophages are viruses that infect bacteria.
- Understanding phage DNA replication is crucial for genetic engineering.
- Lactococcus lactis is a key bacterium in dairy fermentation.
Purpose of the Study:
- To identify and characterize the origin of DNA replication in prolate lactococcal phage c2.
- To investigate the host specificity of this replication origin.
- To develop novel phage-based vectors for Lactococcus lactis.
Main Methods:
- DNA fragment cloning and sequencing.
- Replication assays in Lactococcus lactis and Escherichia coli.
- Sequence comparison with related prolate phages.
- Bioinformatic analysis for open reading frames.
Main Results:
- An origin of DNA replication (ori) was identified in the intergenic region of phage c2.
- The ori fragment directed DNA replication specifically in Lactococcus lactis.
- A conserved nucleotide sequence was found in the ori region across related phages.
- No open reading frames were detected within the ori fragment, suggesting reliance on host factors.
Conclusions:
- A novel origin of DNA replication for lactococcal phage c2 has been characterized.
- The ori exhibits host specificity for Lactococcus lactis.
- The findings facilitate the construction of new lactococcal and shuttle vectors.