Isolation and sequencing of a temperate transducing phage for Pasteurella multocida

Area of Science:

• Microbiology
• Virology
• Genetics

Background:

• Pasteurella multocida is a significant bacterial pathogen.
• Bacteriophages are viruses that infect bacteria and can be used as tools in molecular biology.
• Genetic manipulation techniques are crucial for understanding bacterial pathogens.

Purpose of the Study:

• To isolate and characterize a novel temperate bacteriophage from Pasteurella multocida.
• To determine the genome sequence and host range of the isolated bacteriophage.
• To evaluate the potential of the bacteriophage as a tool for Pasteurella multocida genetic manipulation.

Main Methods:

• Isolation of bacteriophage F108 via mitomycin C induction.
• Morphological characterization using electron microscopy.
• Genome sequencing and bioinformatic analysis.
• Host range determination through infection assays.
• Prophage integration site mapping.
• Transduction experiments using chromosomal markers.

Main Results:

• A temperate bacteriophage, F108, belonging to the Myoviridae family was isolated.
• F108 infects all tested Pasteurella multocida serogroup A strains but not other serotypes.
• The F108 genome is a 30,505-bp double-stranded DNA with cohesive ends, showing homology to Haemophilus influenzae phages.
• The F108 prophage integrates into a tRNA(Leu) gene.
• F108 performs generalized transduction in P. multocida.
• The F108 genome lacks pathogenic genes.

Conclusions:

• Bacteriophage F108 is the first sequenced P. multocida phage and is specific for serogroup A.
• Its ability to perform generalized transduction and lack of pathogenic genes make it a promising tool for P. multocida genetic engineering.
• F108 offers a novel avenue for research and potential therapeutic applications against P. multocida infections.