Genomic Analyses of the Novel Mycobacteriophage KashFlow

Faculty Mentor(s)

Dr. Aaron Best, Biology, Dr. Joseph Stukey

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Eighteen new mycobacteriophages were isolated from soil samples collected around the state of Michigan and parts of the United States. All phages were capable of infecting Mycobacterium smegmatis and were isolated through either enrichment or direct plating at 32°C or 37°C. A variety of plaque morphologies were produced based on size, shape, and clarity; both lytic and temperate phages appear represented in this collection. The mycobacteriophage, KashFlow, was chosen as one of three phages for complete genome sequencing and comparative genomic analyses. The predominant plaque produced by KashFlow at 37°C was circular and 2 mm in diameter. The complete genome sequence for KashFlow revealed relationships to members of the Cluster J, which contains 30 sequenced members to date. The genome of KashFlow is 111.6 Kb, 60.2% GC, and contains 243 genes, including 2 tRNA(Lys-TTT) genes. Upon initial analysis, KashFlow shares a number of gene products with several other phages belonging to the J cluster. In a majority of these instances, these gene products are nearly identical to one another and are almost always perfectly aligned. This trend is predicted to continue throughout the entirety of the KashFlow genome as a number of cluster J phages share these same attributes. A detailed analysis of the complete genome sequences and comparison with sequenced mycobacteriophages is the subject of the second semester of this yearlong course and is presented.


Supported by the Howard Hughes Medical Institute SEA-PHAGES Program.

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