Complete Genome Sequence of Cluster EC Microbacteriophage Piperis
Ryan Kistemaker, Annica Teasley, Lavender Cook, Alyssa Moliis, Anna Josey, Thomas Ray, Farrah Collins, Alisa Whitcomb, Ryan Swarts, Yazmen Roseboro, Shallah Glover, Ian Goodman, Kaitlyn Dishman, Kimberlyn Pickens, Justin Leonard, D. Parks Collins
Microbacteriophage Piperis is a cluster EC bacteriophage which was isolated from soil collected in Mooresville, NC. Piperis has a 53241bp genome with 91 predicted genes.
Microbacteriophages are viruses which are known to infect microbacterial host species. Microbacteriophage Piperis was isolated from soil collected Mooresville, North Carolina, USA, by using Microbacterium foliorumNRRL B-24224 SEA as a host. Piperis was isolated using the enriched isolation procedure at 30ºC in the fall of 2018. Its plaques consisted of numerous small plaques with occasional big plaques and were usually clear. Electron microscopy showed that this bacteriophage has Siphoviridae morphology.
Piperis plaques were selected, purified, and amplified, and the DNA was extracted. The Piperis genome was sequenced with the Illumina Sequencing platform with an approximate shotgun coverage of 43x. The genome was then annotated using the following databases and software: DNA Master, Glimmer, GeneMark, Starterator, Phamerator, PhagesDB.org, NCBI BLAST, and Conserved Domain Database at NCBI, HHPRED, ARAGORN, and PECAAN.
Piperis is a member of Actinobacteriophage cluster EC. The Piperis genome is 53241 base pairs with a circularly permuted genome end and a 68.9% GC content. After annotation, 91 genes were identified, with 32.9% of genes being assigned a function. Piperis was compared to other bacteriophage genomes by using NCBI Blast and was found to be most similar to bacteriophage KaiHaiDragon, with 97% identity over 95% coverage.
Piperis is arranged canonically. The left-hand side has structural genes, while the right-hand side has to do with structural DNA and replication of DNA. This right-hand side begins at gene 48, about halfway through Piperis. The genome ends are circularly permuted. A similar arrangement can be found in the EC cluster, along with KaiHaiDragon, the most similar phage to Piperis. Among the functions in this phage, lysin A can be found upstream from a membrane protein, which could indicate that this is a holin.
When Piperis was compared to other genes in the EC cluster, the phage that is most similar to it is KaiHaiDragon. Most genes in both phages share similar synteny and have the same functions. Neither of these phages has a tRNA gene within their genome. Furthermore, none of the genes in this cluster are read in reverse.
This work was supported by the Howard Hughes Medical Institute (HHMI) Science Education Alliance–Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) program. We thank Graham F. Hatfull, Welkin H. Pope, Deborah Jacobs-Sera, Daniel A. Russell, and Rebecca A. Garlena for their technical support during the sequencing and annotation of this genome.
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