Title of Abstract

Discovery and Characterization of Novel Bacteriophages with Host Bacteria Mycobacterium smegmatis.

Session Title

Poster Session 2

Faculty Mentor

Victoria J. Frost, Ph.D.| Kristi M. Westover Ph.D.

College

College of Arts and Sciences

Department

Biology

Abstract

The Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science program (SEA-PHAGES) is an undergraduate program designed to conduct research on bacteriophages for the purpose of increasing overall bacteriophage knowledge. Antibiotic resistance is a rapidly developing issue due to misuse of antibiotics worldwide. Bacteriophages may provide an alternative treatment solution, given they are highly host-specific. Discovering and characterizing new phages will provide a broader database of candidates for phage therapy. During the fall semester, each student isolated and characterized a soil-found bacteriophage using host bacteriaMycobacterium smegmatis mc2155. Bacteriophages were purified and amplified to provide high titer lysates, or higher phage particle amounts. Genomic DNA from each phage was extracted for restriction enzyme digests followed by gel electrophoresis and possible sequencing. Bacteriophage morphology was determined using transmission electron microscopy (TEM). Of the 13 phages viewed, 12 were of the Siphoviridae morphology (Ashballer, Benson, Blessing, Bombitas, Franica, Gabby, Glaceon, Kallen, Neptr, Pirpich, Scrappy, Thug), and one had a Myoviridae morphology (Jinnie). Unique restriction fragment banding patterns were used to determine which phage DNA samples were sent to Pittsburgh Bacteriophage Institute for sequencing. This sequencing allows the undergraduate students to obtain genomic DNA data for further analysis downstream. Bacteriophages Ashballer (52,231 bp) and Bombitas (110,129 bp) were sequenced and determined to be in sub-clusters A1 and cluster J, respectively. Future work on annotating these genomes, determining location and function of putative genes, is the work of the second component of the SEA program: Bioinformatics.

Course Assignment

BIOL 270 – Frost and Westover

Previously Presented/Performed?

Winthrop University Showcase of Undergraduate Research and Creative Endeavors, Rock Hill, SC, April 2023.

Type of Presentation

Poster presentation

Grant Support?

Supported by HHMI SEA-PHAGES Discovery, Howard Hughes Medical Institute, 2016

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Discovery and Characterization of Novel Bacteriophages with Host Bacteria Mycobacterium smegmatis.

The Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science program (SEA-PHAGES) is an undergraduate program designed to conduct research on bacteriophages for the purpose of increasing overall bacteriophage knowledge. Antibiotic resistance is a rapidly developing issue due to misuse of antibiotics worldwide. Bacteriophages may provide an alternative treatment solution, given they are highly host-specific. Discovering and characterizing new phages will provide a broader database of candidates for phage therapy. During the fall semester, each student isolated and characterized a soil-found bacteriophage using host bacteriaMycobacterium smegmatis mc2155. Bacteriophages were purified and amplified to provide high titer lysates, or higher phage particle amounts. Genomic DNA from each phage was extracted for restriction enzyme digests followed by gel electrophoresis and possible sequencing. Bacteriophage morphology was determined using transmission electron microscopy (TEM). Of the 13 phages viewed, 12 were of the Siphoviridae morphology (Ashballer, Benson, Blessing, Bombitas, Franica, Gabby, Glaceon, Kallen, Neptr, Pirpich, Scrappy, Thug), and one had a Myoviridae morphology (Jinnie). Unique restriction fragment banding patterns were used to determine which phage DNA samples were sent to Pittsburgh Bacteriophage Institute for sequencing. This sequencing allows the undergraduate students to obtain genomic DNA data for further analysis downstream. Bacteriophages Ashballer (52,231 bp) and Bombitas (110,129 bp) were sequenced and determined to be in sub-clusters A1 and cluster J, respectively. Future work on annotating these genomes, determining location and function of putative genes, is the work of the second component of the SEA program: Bioinformatics.