Singers and Soda: The discovery and characterization of A. globiformis phages Pitbull, MissAmericana, RootBeer, DoctorPepper, and MellowYellow

Abby Antonishek, Isabella Belperio, Taylor Buchowski, Adelaide Bullard, Alexis Carroll, Umbert Caseres, Faith Chapman, Sarah Choi, Ashley Cumbee, Priyanshu Datta Roy, Luke DeLion, Katherine DeMarco, Armand Destin, Aidan Dibble, Anabela Djurovic-Topalovic, Lucy Ehmann, Elizaveta Filippova, Anagha Gaitonde, Junezuh Ghang, Alexandra Gilman, Kayla Grennes, Zion Hackett, Salem Hallab, Natalie Haynes, Carl Henry, Zachery Heppner, Alex Higgins, Ryan Houser, Alyssa Hudgins, Emma Jefferies, Jingyi Ji, Seth Jimenez, Emma Kessinger, Carolina Kim, Morgan Laskowski, Ryan Lee, Manasvi Lingampally, Alex Martin, Stan Melkumian, Aishwarya Menon, Gio Minghetti, Amy Murdock, Henrietta Ndubuokwu, Thanh Nguyen, Alyson Owen, Megan Painter, Amanda Pawlecki, Michael Perrin, Dane Phillips, Alex Pray, Amalia Rosenthal, Prakhar Saxena, Hudson Schneider, Laney Shafer, Bradley Shannon, Ethan Shatz, Molly Shean, Katie Smith, Adalene Stangeland, Kate Stanton, Maddie Stevenson, Gianna Stinsa, Dean Stornello, Drake Strait, Derek Temple, Lauren Thompson, Grace Troyer, Ann Uhlmansiek, Lucas Urquiaga, Faith Wagner, Adam Weber, Tristen West, Amelia Winkelman, Amanda Wolf, Lillian Woolley, Danae Youngstedt, Joyce Yu, Conwy Zheng, Irina Zhilinskaya, Nicholas Zolnierczuk

In the Fall of 2023 five bacteriophages Pitbull (FQ), MissAmericana (EM2), Rootbeer (FA), DoctorPepper (AW), and MellowYellow (AP2) were sequenced out of 46 discovered phages isolated by Purdue University undergraduates using A. globiformis B-2979. These bacteriophages showed notable morphological and genetic diversity from each other as well as previously sequenced phages with the same bacterial host. Of the five sequenced phages, all were siphoviridae, except MissAmericana which was podoviridae. However, Pitbull, MissAmericana, and Rootbeer all exhibited lysogenic properties during initial isolation in which they developed bacterial colonies inside of phage plaques. Their genome size and numbers of genes had a range of approximately 39 to 73 kbp and 51 to 125 genes, respectively. Pitbull, MissAmericana, and MellowYellow all had less than five other members in their (sub)clusters, indicating especially novel genomes. Approximately 60% of genes in MissAmericana and MellowYellow were reverse genes. Additionally, 20% of Pitbull’s genes were orphams, indicating a high degree of gene novelty. Select genes and gene regions from the five bacteriophages were further investigated using a combination of AlphaFold, PyMol, Phamerator, HHPred, PhagesDB, custom Python scripts, and DeepTMHMM.