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A bacteriophage is a virus capable of infecting bacteria like ubiquitous soil-dwelling genus Bacillus. Within the Bacillus genus, there is the “ACT family” made up of B. thuringiensis, B. cereus, and B. anthracis, which are highly related but with different pathogenic characteristics. Because of this, phages isolated using a species in this group may have a broad host range encompassing several species from Bacillus. Since B. cereus and B. anthracis can result in fatal to mild sickness in humans, the non-pahtogenic B. thuringiensis kurstaki was used to discover and characterize novel phages. The phage OTooleKemple52 was isolated from a soil sample collected from Chesapeake, VA using “soil enrichment” to increase phage concentration and thus make detection more likely. A phage plaque was observed from this enrichment infection upon plating. The phage population was then purified until the morphology of the phage plaques was consistent (3 mm diameter with pinpoint clear centers). A large volume of phage stock (high titer lysate, HTL) was collected and was then used to obtain purified DNA for gel electrophoresis and genome sequencing. Additionally, an HTL sample was stained with 1% uranyl acetate and imaged using transmission electron microscopy to determine a myoviridae morphology with a head diameter of 80 nm and a tail length of 200 nm. The phage has a broad host range and was able to able to form plaques on 6 out of 8 Bacillus strains tested. The genome of the phage will be annotated and compared to other phage genomes.
Through studying phages we can work to better understand both phage diversity and the interrelatedness of the Bacillus genus. The benefits of studying bacteriophages have reaches from environmental to medical significance because of the ubiquitous and pathogenic characteristics of the host bacteria.
Bacillus thuringiensis, bacteriophage, phage, bioinformatics, SEA-PHAGES
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