32. Isolation and Characterization of the Novel Bacteriophage LilHazelnut

Abstract

Bacteriophages (phage) make up a diverse and abundant population of viruses that replicate by hijacking the cellular machinery of a bacterial host. Only a minute fraction of the phage population is characterized phenotypically or genetically, and to date, this small percentage represents a large diversity categorized into 88 clusters and 73 subclusters. Given the potential of these phages, in our understanding of evolution, biological diversity, and especially medical applications, such as vaccine and diagnostic development for the growing prevalence of antibiotic resistant bacterial pathogens, we predict that by isolating and genetically characterizing a novel phage we will be able to contribute to this growing scientific field. We commenced our research by using Mycobacterium smegmatis, a non-pathogenic bacterial host, in order to isolate an individual phage, LilHazelnut, from a soil sample through a series of isolation, amplification, and purification protocols. This was done in order to yield an adequate pure titer of a single phage. TEM microscopy determined that this particular phage is a short-tailed Siphoviridae. The DNA of this phage was then extracted, and its genome was sequenced, which identified 87 predicted genes and clearly demonstrated it as a novel member of the highly conserved Q cluster of phages. Through the ongoing annotation of LilHazelnut’s genome, we hope to discover novel genes and expand on what we have already uncovered about the unique Q cluster while illustrating the importance of genomically characterizing and cataloguing phage.

Keywords: phage, isolation, genome, annotation, Mycobacterium smegmatis