During the 2016-2017 academic year, students isolated, purified, and characterized 11 bacteriophages found in the soil in and around Winthrop University, Rock Hill, South Carolina. See the Actinobacteriophage Database for individual phage information.
Each phage was amplified in the bacterial host Mycobacterium smegmatis mc2155 and then characterized following DNA extraction using restriction enzyme digests and gel electrophoresis. Electron microscopy demonstrated that all eleven phages had long, flexible tails and belong to the Siphoviridae group of mycobacteriophages.
After studying the electron micrographs, the digestion patterns, and quality of the DNA, two of the most potentially unusual phages, Asriel and HaiMas, were chosen to be sequenced at Pittsburg State University (PSU). In the following semester, research continued and both genomes were annotated using DNA Master Software and several homology search programs, including BLASTp and HHPred, to predict gene locations and determine gene function.
Asriel is 74,594 bp in length, has approximately 142 predicted genes, and is a member of Cluster E. Similar to other Cluster E phages, the Asriel genome codes for two tRNAs, and a putative immunity suppressor protein. HaiMas is 68,296 bp in length, has approximately predicted 99 genes, and is a member of Cluster B1. Both genomes encode for integrase proteins which suggest a temperate lifestyle. These genomes are ready to be submitted for peer review and ultimately deposited at the National Library of Medicine's database, GenBank. We are also preparing a manuscript for Genome Announcements.
Students presented their work at the Association for Microbiology Society's regional meeting at Francis Marion University, Florence, South Carolina. They also participated in Winthrop University's annual undergraduate research conference (SOURCE) and at the Department of Biology's annual spring research showcase. In June 2017, two students will present the work at HHMI's national SEA-PHAGES conference in Washington, DC.
