Genetic Analysis of Novel Mycobacteriophages: SheaKeira and Adonis
Abstract
Genetic Analysis of Novel Mycobacteriophages: SheaKeira and Adonis Keira Stacks, Shea Morris and Indiren Pillay Mycobacteriophages are a genetically diverse set of viruses that can infect large amounts of mycobacteria hosts such as Mycobacterium smegmatis and Mycobacterium tuberculosis. These viruses are abundant throughout the biosphere and have been evolving for billions of years, giving rise to highly diverse populations. This study investigates the genetic makeup of mycobacteriophages to grasp a deeper understanding of their molecular biology and the overall evolution of bacteriophages. A non-pathogenic host, Mycobacterium smegmatis, was used to isolate phages from soil samples collected at Lake Laurel and Oconee River Greenway in Milledgeville, Georgia. The purified phage lysates were imaged by transmission electron microscopy. Electron micrographs showed that both phages have icosahedral capsids with non-contractile tails, but appear to be different organisms, subsequently named SheaKeira and Adonis. Phage DNA was then extracted, purified, and characterized by restriction endonuclease digestion with BamHI, Clal, EcoRI, HaeIII and HindIII. Illumina sequencing of Adonis and SheaKeira DNA was conducted and the genome sequence compared to an existing online phage database. Phage Adonis, showed similarity to subcluster K1 phages and SheaKeira appears to belong to phage subcluster A9. Furthermore, a DNA analysis software program was used to annotate the genomic sequence of these two phages to increase our knowledge of the genetic framework of these diverse groups of bacteriophages.
Genetic Analysis of Novel Mycobacteriophages: SheaKeira and Adonis
Genetic Analysis of Novel Mycobacteriophages: SheaKeira and Adonis Keira Stacks, Shea Morris and Indiren Pillay Mycobacteriophages are a genetically diverse set of viruses that can infect large amounts of mycobacteria hosts such as Mycobacterium smegmatis and Mycobacterium tuberculosis. These viruses are abundant throughout the biosphere and have been evolving for billions of years, giving rise to highly diverse populations. This study investigates the genetic makeup of mycobacteriophages to grasp a deeper understanding of their molecular biology and the overall evolution of bacteriophages. A non-pathogenic host, Mycobacterium smegmatis, was used to isolate phages from soil samples collected at Lake Laurel and Oconee River Greenway in Milledgeville, Georgia. The purified phage lysates were imaged by transmission electron microscopy. Electron micrographs showed that both phages have icosahedral capsids with non-contractile tails, but appear to be different organisms, subsequently named SheaKeira and Adonis. Phage DNA was then extracted, purified, and characterized by restriction endonuclease digestion with BamHI, Clal, EcoRI, HaeIII and HindIII. Illumina sequencing of Adonis and SheaKeira DNA was conducted and the genome sequence compared to an existing online phage database. Phage Adonis, showed similarity to subcluster K1 phages and SheaKeira appears to belong to phage subcluster A9. Furthermore, a DNA analysis software program was used to annotate the genomic sequence of these two phages to increase our knowledge of the genetic framework of these diverse groups of bacteriophages.