Presenters: Parker Folsom ; Kirsten Abbott ; Sam Aldous
Authors: Parker Folsom, Kirsten Abbott, Sam Aldous, Stephen Adams
Faculty Advisor: Julianne Grose
Institution: Brigham Young University
> > Due to the growing threat of antibiotic resistance and recent experimental bacteriophage therapies, a great deal of research has recently been performed on bacteriophages that target various ESKAPE pathogens and their close relatives. Before a bacteriophage can be used in clinical settings, it is important to identify the function of its proteins to ensure its safety. Additionally, it is important to determine which clinical isolates it will be effective against. Herein we describe the isolation and characterization of a bacteriophage that infects Klebsiella aerogenes, the K in ESKAPE. The bacteriophage was isolated from sewage samples, brought to titer, sequenced, and imaged using an electron microscope. Genomic annotation was completed using the DNAmaster and Genemark software. Unfortunately, genomic analysis relying solely on computer programs may miss unique/novel proteins, and doesn’t provide information on protein expression levels. We hypothesized that mass spectrometry can be used to identify previously uncalled proteins and will show expression levels for some of the most common proteins. After performing mass spectrometry, it was determined that computer-based annotation had failed to identify, or had not correctly called the start of several proteins. It was also determined that mass spectrometry may be used as a simple way of determining protein expression levels. These results suggest that traditional methods for genome annotation of bacteriophages are a good start, but could miss some proteins, especially if those proteins are novel.