Investigating Pepperwood: An In-Depth Analysis of a Novel Bacteriophage
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Presenter(s)
Braeden Rose, Anushka Viswanathan, Shreya Chilukuri, Murali Devanaboyina, Soumya Rao
Faculty Research Mentor
Dr. Kathleen Hafer and Dr. Christopher Shaffer
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No
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Acknowledgments
We would like to thank Professors Dr. Kathleen Hafer and Dr. Christopher Shaffer for their guidance and for overseeing WashU's involvement in the national SEA-PHAGES program. We would also like to thank the Howard Hughes Medical Institute for their funding of this program.
Abstract or Description
The characterization and analysis of a novel phage was performed by undergraduate researchers as part of the SEA-PHAGES program. Each phage discovered and annotated becomes part of a growing database that contributes to scientific understanding of phage biodiversity. Phage Pepperwood is a newly isolated phage of the bacteria Streptomyces lividans. It was isolated from a soil sample collected in Pittsburg, Missouri. Pepperwood forms small, circular plaques with well-defined edges when grown on its bacterial host. Based on EM studies, Pepperwood is a siphoviridae characterized by an icosahedral head and long tail. Pepperwood’s genomic sequence was completed over winter break. Genetic analysis revealed it is a member of the BE1 subcluster. Pepperwood has a GC content of 50.1%, a length of 133,012 base pairs, and contains 229 genes. These statistics indicate that this phage is similar to other phages in the cluster, deviating by an average 0.8%. The phage contains a long terminal repeat consisting of 24 genes and 11,227 base pairs, meaning the phage contains only 205 unique genes. Pepperwood’s closest relatives are the phages Sushi23, Bartholomune, and Peebs, all members of the BE1 subcluster. Of the 205 unique genes, 88 are classified as no known function, 12 genes have been assigned putative functions, and 31 have confirmed protein functions. 74 genes remain uncharacterized. Pepperwood codes for 45 tRNAs while the average for the subcluster is 44.3. Gene 47 codes for Pepperwood’s portal protein. The phage contains a tail assembly chaperon located at gene 56, however no slippery sequence has been found within the genome nor in any closely related phage genomes. In the future, Phage Pepperwood could be tested against other bacterial hosts to determine the extent of its virulence and its applicability to the field of phage therapy.