Isolation of 35 Mycobacteriophages and Genomic Analysis of the Novel Mycobacteriophage, Bella96
Dr. Aaron Best and Dr. Joseph Stukey
Thirty-five new mycobacteriophages were isolated from soil samples collected on or nearby Hope College in Holland, Michigan. All were capable of infecting Mycobacterium smegmatis and produced a variety of plaque morphologies based on size, shape, and clarity, consistent with the isolation of an assortment of different phages. Both lytic and temperate phages appear represented in this collection. Purified phage stocks were used to prepare genomic DNA samples for restriction digest analysis. A comparison of those 35 digest results revealed few similarities among the group, further supporting our interpretation that most of the new phage isolates were distinct. One mycobacteriophage, Bella96, was chosen for complete genome sequencing using the Illumina MiSeq platform and comparative genomic analysis. The predominant plaque produced by Bella96 at 32⁰C was turbid and 3 mm in diameter. No plaques grew at 42⁰C. Genome sequence data for Bella96 revealed a relationship to a group of 23 mycobacteriophages in Cluster K1. The genome of Bella96 is 60.7 Kb, 66.1% GC, and contains 97 genes in agreement with the genome characteristics of closely related phage. A detailed analysis of the complete genome sequence and comparison with sequenced members of this small and unique group of mycobacteriophages is the subject of the second semester of this yearlong course and is presented.
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