Western Kentucky University

Poster Title

Genomic Characterization of Mycobacteriophage TiroTheta9

Institution

Western Kentucky University

Abstract

Bacteriophages are the most abundant biological entities on earth, and yet very few types have been characterized. It is imperative that we analyze as many types as possible to gain important insights into molecular biology, genetics, evolution, and even pharmaceutics as researchers begin to recognize the potential of phage as a replacement for antibiotics. The objective of this project was to isolate and characterize a novel bacteriophage from the environment. Mycobacterium smegmatis, a harmless bacterium commonly found in soil, served as the host and facilitated the enrichment and recovery of mycobacteriophages. A single phage type was purified to homogeneity and was designated TiroTheta9. The morphology of the purified TiroTheta9 phage particles was visualized by electron microscopy. TiroTheta9 genomic DNA was isolated and analyzed using DNA restriction enzymes and gel electrophoresis before being sequenced. The sequenced genome was annotated and compared to the genomes of other mycobacteriophages. This analysis revealed that TT9 shares significant sequence similarity to mycobacteriophage Peaches. Based on these data, it was concluded that TiroTheta9 is a unique mycobacteriophage with close similarity to Peaches and other mycobacteriophages of the A4 subcluster. The genomic information gathered on TT9 will add to the growing database of annotated phage genomes and serve to augment our understanding of bacteriophages in general.

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Genomic Characterization of Mycobacteriophage TiroTheta9

Bacteriophages are the most abundant biological entities on earth, and yet very few types have been characterized. It is imperative that we analyze as many types as possible to gain important insights into molecular biology, genetics, evolution, and even pharmaceutics as researchers begin to recognize the potential of phage as a replacement for antibiotics. The objective of this project was to isolate and characterize a novel bacteriophage from the environment. Mycobacterium smegmatis, a harmless bacterium commonly found in soil, served as the host and facilitated the enrichment and recovery of mycobacteriophages. A single phage type was purified to homogeneity and was designated TiroTheta9. The morphology of the purified TiroTheta9 phage particles was visualized by electron microscopy. TiroTheta9 genomic DNA was isolated and analyzed using DNA restriction enzymes and gel electrophoresis before being sequenced. The sequenced genome was annotated and compared to the genomes of other mycobacteriophages. This analysis revealed that TT9 shares significant sequence similarity to mycobacteriophage Peaches. Based on these data, it was concluded that TiroTheta9 is a unique mycobacteriophage with close similarity to Peaches and other mycobacteriophages of the A4 subcluster. The genomic information gathered on TT9 will add to the growing database of annotated phage genomes and serve to augment our understanding of bacteriophages in general.