--- id: "edwards-2023-mraaronian" title: "Complete genome sequence of bacteriophage MrAaronian isolated from an Arthrobacter globiformis culture" authors: - "Ian Haines" - "Jessica Blakely" - "Ashley Branson" - "Diana Estrada" - "Rebeca Fernandez Robles" - "Katelyn Fitzgerald" - "Shelby Jeffers" - "Timyee Leung" - "Jasmine Munoz" - "Ashley Olivos" - "Anayeli Ramirez" - "Caressa Smith" - "Justin Spere" - "Idaleth Tavarez" - "Isabella Wood" - "Ethan Zavala" - "Madison Arrighi" - "Angelica Coronel-Galindo" - "Megan K. Dennis" - "Marlee Goppert" - "Dustin Edwards" venue: "Microbiology Resource Announcements" year: 2023 date: "2023-12-14" doi: "10.1128/MRA.00778-23" url: "/research/publications/10-1128-mra-00778-23/" pdf: "/research/publications/10-1128-mra-00778-23/dustin-edwards-10-1128-mra-00778-23.pdf" pmc: "https://pmc.ncbi.nlm.nih.gov/articles/PMC10720457/" openAccess: true license: "cc-by" accessions: - "genbank:OR159662" - "sra:SRX21368075" citedBy: 0 citedBySource: "OpenAlex, read 2026-09-12" --- # Complete genome sequence of bacteriophage MrAaronian isolated from an Arthrobacter globiformis culture Genome of phage MrAaronian, isolated from flowerbed soil in Poughkeepsie; 54,509 bp, 87 genes, cluster AW. ## Abstract Arthrobacteriophage MrAaronian contains a 54,509 bp DNA genome with 87 predicted protein-coding genes. MrAaronian has siphovirus morphology and was collected from a flowerbed soil sample in Poughkeepsie, NY, and isolated on an Arthrobacter globiformis B-2979 culture. MrAaronian has > 99% nucleotide identity with cluster AW arthrobacteriophages Michelle, Stayer, Sloopyjoe, and StarLord. ## Full text Machine-extracted from the PDF linked above. It carries the artifacts that come with reading a typeset two-column page: running heads, figure captions in the flow of the prose, and words broken across line ends. The abstract above is the registry's deposit and is the authoritative text. | Bacteriophages | Announcement Complete genome sequence of bacteriophage MrAaronian isolated from an Arthrobacter globiformis culture Ian Haines,1 Jessica Blakely,1 Ashley Branson,1 Diana Estrada,1 Rebeca Fernandez Robles,1 Katelyn Fitzgerald,1 Shelby Jeffers,1 Timyee Leung,1 Jasmine Munoz,1 Ashley Olivos,1 Anayeli Ramirez,1 Caressa Smith,1 Justin Spere,1 Idaleth Tavarez,1 Isabella Wood,1 Ethan Zavala,1 Madison Arrighi,2 Angelica Coronel-Galindo,2 Megan K. Dennis,2 Marlee Goppert,1 Dustin Edwards1 AUTHOR AFFILIATIONS See affiliation list on p. 3. ABSTRACT Arthrobacteriophage MrAaronian contains a 54,509 bp DNA genome with 87 predicted protein-coding genes. MrAaronian has siphovirus morphology and was collected from a flowerbed soil sample in Poughkeepsie, NY, and isolated on an Arthrobacter globiformis B-2979 culture. MrAaronian has > 99% nucleotide identity with cluster AW arthrobacteriophages Michelle, Stayer, Sloopyjoe, and StarLord. KEYWORDS bacteriophages, Arthrobacter The Arthrobacter group of bacteria is unusual since they appear as Gram-negative rods in young cultures and as Gram-positive cocci in older cultures (1, 2). Arthro bacter globiformis, a soil-dwelling bacterium with potential commercial importance, is able to degrade several pollutants, including hexavalent chromium (3). Further characterization of bacteriophages and viruses that infect specific host bacteria, targeting Arthrobacter could have applications in selective bioremediation. Arthrobacteriophage MrAaronian was isolated from a moist soil sample in Pough keepsie, NY [global positioning system (GPS) coordinates: 41.72201 N, 73.930158 W]. The sample was washed with peptone-yeast calcium (PYCa) liquid media, and the superna tant was passed through a 0.22 μm syringe filter. The filtrate was incubated with host A. globiformis B-2979 in a soft agar overlay on PYCa agar plates at 30°C. Bacteriophages forming small, lytic plaques were isolated by two rounds of picking a single, well-sep arated plaque, followed by diluting the bacteriophage sample in a 10-fold dilution series and plating again with A. globiformis (Fig. 1A). High-titer lysates were prepared by flooding “webbed” plates with phage buffer [10 mM Tris (pH 7.5), 10 mM MgSO4, 68 mM NaCl, 1 mM CaCl2, 10% glycerol], as described in the Phage Discovery Guide (4). Negative-staining transmission electron microscopy showed a siphovirus morphology, and ImageJ v1.53m (5) measured an approximate tail length of 285 nm and capsid diameter of 67.5 nm (Fig. 1B). Bacteriophage genomic DNA was extracted using a Promega Wizard DNA Clean-Up System (Promega, Madison, WI), and sequencing libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit (New England Biolabs, Ipswich, MA). Libraries were sequenced by Illumina MiSeq at the Pittsburgh Bacteriophage Institute to generate 607,351 total single-end reads of 150-base read length. A single bacteriophage contig with 6,670× coverage was assembled using Newbler v2.9 and checked for completeness and genomic termini using Consed v29 (6, 7). The 54,509 bp double-stranded DNA genome contains a 9-nucleotide 3′ single-stranded overhang (5′-CGCCGACCT- 3’) and 51.7% G + C content. BLASTn (8) query of the sequence with the nonredundant/nucleotide (nr/nt) database returned >99% nucleotide sequence identity to cluster AW bacteriophages Michelle, Stayer, Sloopyjoe, and StarLoad (Table 1). Auto-annotations by GLIMMER v3.02 (9) December 2023 Volume 12 Issue 12 10.1128/MRA.00778-23 1 Editor John J. Dennehy, Department of Biology, Queens College, Queens, New York, USA Address correspondence to Dustin Edwards, dcedwards@tarleton.edu. The authors declare no conflict of interest. See the funding table on p. 3. Received 23 August 2023 Accepted 29 September 2023 Published 7 November 2023 Copyright © 2023 Haines et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Downloaded from https://journals.asm.org/journal/mra on 26 July 2026 by 156.146.253.207. and GeneMark v2.5p (10) were manually refined using Phamerator (11), PECAAN (https://discover.kbrinsgd.org), and DNA Master v5.23.6 (http://phagesdb.org/DNAMas ter/). Consistent with other cluster AW bacteriophages, no tRNA genes were identi fied using Aragorn v1.1 (12) and tRNAscan-SE v2.0 (13). TmHmm and SOSUI v1.11 (14) assessed potential trans-membrane helices for each gene product. The genome contained rightward transcribing genes and putative functions was assigned to 25 of 87 protein-coding genes using BLASTp v2.13 (8) and HHPred v3.3 (15), which include structural proteins, endolysin, DNA primase/polymerase, DNA helicase, ssDNA and dsDNA binding proteins, four membrane proteins, two HNH endonucleases, VRR-Nuc domain protein, and Cas4 exonuclease. Gene position and length of MrAaronian were FIG 1 Characterization of arthrobacteriophage MrAaronian. (A) Petri dish (90 mm) containing PYCa solid culture medium and bacterial host Arthrobacter globiformis B-2979 infected with MrAaronian. After incubation for 24 h at 30°C, plaques appeared to be <1 mm in diameter and clear, with no visible halo (scale bar = 10 mm). (B) Transmission electron micrograph of MrAaronian high-titer lysate negatively stained with 1% uranyl acetate on a 300-mesh copper grid and imaged with a Hitachi HT7800 120 kV transmission electron microscope. MrAaronian has a siphovirus morphology with an approximate tail length of 285 nm and a capsid diameter of 67.5 nm (scale bar = 200 nm). (C) A density plot showing the relationship between length and position of genes in cluster AW bacteriophages and MrAaronian (white circles). TABLE 1 Characteristics of similara arthrobacteriophages in cluster AW Phage name Length of genome (bp) G+C content (%) GenBank accession number Similarity to MrAaronian (%) MrAaronian 54,509 51.7 OR159662 Michelle 54,509 51.7 MN234210 99.71 Stayer 54,507 51.7 MN234175 99.37 Sloopyjoe 54,511 51.7 OP910131 99.24 StarLord 54,504 51.7 MN234229 99.15 aSimilar being defined as having a nucleotide similarity over 95%. Announcement Microbiology Resource Announcements December 2023 Volume 12 Issue 12 10.1128/MRA.00778-23 2 Downloaded from https://journals.asm.org/journal/mra on 26 July 2026 by 156.146.253.207. compared against other cluster AW members in a density plot using observable (16) showing three gene groupings (Fig. 1C). All tools were run with default parameters. ACKNOWLEDGMENTS We thank Graham Hatfull, Deborah Jacobs-Sera, Daniel Russell, Rebecca Garlena, and Richard Pollenz for their technical support during the sequencing and annotation of this genome. Support for this research was provided by the Tarleton State University College of Science and Mathematics, Marist College School of Science, and the Howard Hughes Medical Institute Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) research and education program. AUTHOR AFFILIATIONS 1Department of Biological Sciences, Tarleton State University, Stephenville, Texas, USA 2Department of Biology, Marist College, Poughkeepsie, New York, USA AUTHOR ORCIDs Dustin Edwards http://orcid.org/0000-0001-6409-8041 FUNDING Funder Grant(s) Author(s) Tarleton State University College of Science and Mathematics Dustin Edwards AUTHOR CONTRIBUTIONS Ian Haines, Formal analysis, Visualization, Writing – original draft | Jessica Blakely, Formal analysis, Writing – original draft | Ashley Branson, Formal analysis, Writing – original draft | Diana Estrada, Formal analysis, Writing – original draft | Rebeca Fernandez Robles, Formal analysis, Writing – original draft | Katelyn Fitzgerald, Formal analysis, Writing – original draft | Shelby Jeffers, Formal analysis, Writing – original draft | Timyee Leung, Formal analysis, Writing – original draft | Jasmine Munoz, Formal analysis, Writing – original draft | Ashley Olivos, Formal analysis, Writing – original draft | Anayeli Ramirez, Formal analysis, Writing – original draft | Caressa Smith, Formal analysis, Writing – original draft | Justin Spere, Formal analysis, Writing – original draft | Idaleth Tavarez, Formal analysis, Writing – original draft | Isabella Wood, Formal analysis, Writing – original draft | Ethan Zavala, Formal analysis, Writing – original draft | Madison Arrighi, Formal analysis, Investigation | Angelica Coronel-Galindo, Formal analysis, Investigation | Megan K. Dennis, Data curation, Formal analysis, Funding acquisition, Investigation, Methodol ogy, Project administration, Supervision, Visualization | Marlee Goppert, Formal analysis, Supervision, Writing – original draft | Dustin Edwards, Data curation, Formal analy sis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Visualization, Writing – original draft DATA AVAILABILITY Actinobacteriophage MrAaronian genome is available in GenBank as accession number OR159662. Raw reads are available in the SRA under accession number SRX21368075. REFERENCES 1. Conn HJ. 1928. A type of bacteria abundant in productive soils, but apparently lacking in certain soils of low productivity. Cornell Univ. 2. Jones D, Keddie RM. 2006. The genus arthrobacter. Prokaryotes 3:945– 960. https://doi.org/10.1007/0-387-30743-5 3. Megharaj M, Avudainayagam S, Naidu R. 2003. 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