Annotation and Host Range Analysis of Bacteriophage Statler
Faculty Mentor Information
Dr. Micheal A. Thomas (Mentor), Idaho State University; and Dr. Jack Shurley (Mentor), Idaho State University
Abstract
Bacteriophage Statler, infecting Microbacterium foliorum NRRL B-24224, was isolated in 2023 by ISU BIOL 1111 Phage Discovery Lab (part of HHMI’s SEA-PHAGES program) from a thatch & soil sample. Statler belongs to actinobacteriophage cluster EG and has siphovirus morphology. Following DNA sequencing and assembly, I used bioinformatics tools to identify 40 functional proteins out of 110 genes. Notably, Statler contains a new function in Microbacterium phages called cyclic oligonucleotide sequestration protein, Acb2; this protein (gp57) is involved in inhibition of bacterial antiphage CBASS gene. Additionally, there is a probable holin protein (gp66) that degrades host cell walls, allowing newly constructed phages to escape. A host range analysis was performed with phages Statler, Rowlf (cluster EG), SallyK (EG) and KillerTomato (EE), all found on ISU campus by previous BIOL 1111 students. These were tested for their ability to infect hosts other than M. foliorum, using 12 Microbacterium host species and 4 other bacterial species (Staphylococcus aureus, E. coli, Bacillus subtilis & B. cereus) in a series of spot tests and plaque assays. All M. foliorum phages have a narrow host range, infecting only 1 or 2 additional Microbacterium species and none of the other genera, with little overlap among preferred hosts.
Annotation and Host Range Analysis of Bacteriophage Statler
Bacteriophage Statler, infecting Microbacterium foliorum NRRL B-24224, was isolated in 2023 by ISU BIOL 1111 Phage Discovery Lab (part of HHMI’s SEA-PHAGES program) from a thatch & soil sample. Statler belongs to actinobacteriophage cluster EG and has siphovirus morphology. Following DNA sequencing and assembly, I used bioinformatics tools to identify 40 functional proteins out of 110 genes. Notably, Statler contains a new function in Microbacterium phages called cyclic oligonucleotide sequestration protein, Acb2; this protein (gp57) is involved in inhibition of bacterial antiphage CBASS gene. Additionally, there is a probable holin protein (gp66) that degrades host cell walls, allowing newly constructed phages to escape. A host range analysis was performed with phages Statler, Rowlf (cluster EG), SallyK (EG) and KillerTomato (EE), all found on ISU campus by previous BIOL 1111 students. These were tested for their ability to infect hosts other than M. foliorum, using 12 Microbacterium host species and 4 other bacterial species (Staphylococcus aureus, E. coli, Bacillus subtilis & B. cereus) in a series of spot tests and plaque assays. All M. foliorum phages have a narrow host range, infecting only 1 or 2 additional Microbacterium species and none of the other genera, with little overlap among preferred hosts.