Genomic Analyses of the Novel Mycobacteriophage Opia

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, Opia, was chosen as one of three phages for complete genome sequencing and comparative genomic analyses. The predominant plaque produced by Opia at 37°C was circular and approximately 1 mm in diameter. The complete genome sequence for Opia revealed relationships to members of the Cluster B2, which contains 22 sequenced members to date. The genome of Opia is 67.4 Kb, 68.9% GC, and contains 93 genes. Phages in Cluster B2 share very high sequence identity throughout the genomes lengths. Opia maintains this pattern, being nearly identical to phages Glass, Rosebush, and Hedgerow despite isolation of the phages in different years (15 year span) and in different geographical locations. 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