BRED: A Tool for Making Mutant Bacteriophage Genomes
Author Information
Author(s): Marinelli Laura J., Piuri Mariana, Swigoňová Zuzana, Balachandran Amrita, Oldfield Lauren M., van Kessel Julia C., Hatfull Graham F.
Primary Institution: Pittsburgh Bacteriophage Institute and Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America
Hypothesis
Can Bacteriophage Recombineering of Electroporated DNA (BRED) be used to efficiently construct targeted mutants of bacteriophage genomes?
Conclusion
The BRED method allows for the efficient construction of various mutations in bacteriophage genomes without the need for selection markers.
Supporting Evidence
- BRED allows for the construction of unmarked gene deletions and precise gene replacements.
- The method can be applied to all tested mycobacteriophages.
- Mutants can be readily detected by PCR without the need for selection.
Takeaway
BRED is a new way to change the DNA of viruses that infect bacteria, making it easier to study how these viruses work.
Methodology
The BRED method involves co-electroporating phage DNA and a targeting substrate into Mycobacterium smegmatis cells to create mutants.
Limitations
The efficiency of recombineering can vary, and some mutants may not be viable.
Digital Object Identifier (DOI)
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