TABLE 1.
Properties of four mycobacteriophages isolated from soil samples collected on August 23, 2022 in Longview, Texas, USA
| Data for mycobacterium phage a | ||||
|---|---|---|---|---|
| Characteristic | Diminimus | Dulcita | Glaske16 | Koreni |
| Soil sampling location GPS coordinates | 32.46474 N , 94.7272 W | 32.464444 N , 94.7275 W | 32.465 N , 94.727778 W | 32.463333 N , 94.726389 W |
| Lysate titer (PFU/mL) | 1.8 × 1,010 | 2.0 × 1,010 | 4.9 × 109 | 1.4 × 1,010 |
| Plaque morphology after 48 h at 37°C | Clear with defined edges | Clear with defined edges | Clear with defined edges | Clear with defined edges |
| Avg plaque diameter (mm [n-value]) | 0.8 (15) | 1.0 (10) | 1.7 (3) | 1.0 (13) |
| Approx. shotgun coverage (X) | 338 | 479 | 360 | 1,490 |
| Genome size (bp) | 80,037 | 80,038 | 81,156 | 51,055 |
| GC content (%) | 61.6 | 61.6 | 61.6 | 63.9 |
| Overhang sequence | ACCTCCTGCAA | ACCTCCTGCAA | ACCTCCTGCAA | CGGCCGGTAA |
| Overhang length (bases) | 11 | 11 | 11 | 10 |
| Cluster | M | M | M | A |
| Subcluster | M1 | M1 | M1 | A4 |
| GenBank accession no. | OR521083 | OR553916 | OR553909 | OR553901 |
| SRA accession no. | SRX19690831 | SRX19690832 | SRX19690837 | SRX19690842 |
| Total no. of reads | 185,644 | 258,101 | 201,637 | 528,548 |
| No. of predicted genes | 137 | 137 | 140 | 90 |
| No. of predicted tRNAs | 19 | 19 | 18 | 0 |
| tRNA type(s) | Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, Val | Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, Val | Trp, Asn, Gln, Tyr, Pro, Ser, Phe, Met, Arg, His, Leu, Lys, Gly, Val, Thr, Asp, Glu | – |
| No. of genes with predicted functions | 51 | 52 | 54 | 48 |
| % of genes with predicted functions | 37.2 | 38 | 38.6 | 53.3 |
| Key predicted lysogenic life cycle genes | Serine integrase | Serine integrase | Serine integrase | Serine integrase, immunity repressor |
| No. of orphams | 0 | 0 | 1 | 0 |
| Avg capsid size (nm [n-value]) | 61.53 (10) | 60.75 (10) | 68.21 (3) | 61.56 (10) |
| Avg tail length (nm [n-value]) | 323.51 (10) | 333.00 (11) | 328.30 (3) | 131.60 (10) |
| Isolated by | Faith W Baliraine, Taryn M Pledger, Camila Andrade, Chloe I Wade | Krista L Anderson,Olivia L Donnelly,Hannah L Foree, Kaitlyn E Mathews | Sofia N Mendez, Tadeen Feroz, Amya R Orn, Jerron Hudson | Krista L Anderson,Olivia L Donnelly,Hannah L Foree, Kaitlyn E Mathews |
All bacteriophages were isolated using the enriched isolation method [reference (10)] and purified through three sequential (37°C, 48 h) rounds of plating with Mycobacterium smegmatis mc2155 cells in Middlebrook 7H9 top agar. Genome sequencing was accomplished using the Illumina Shotgun sequencing method with 150-base single-end reads using the NEB Ultra II Library sequencing kit (v3 reagents). All had 3′ single-stranded overhang genome ends. Genomic termini were identified through buildups of read start positions and variations in genome-wide coverage and manually verified using Consed version 29 [references (14) and (15) ]. All bacteriophages had a siphovirus morphotype and were predicted to be lysogenic based on the presence of predicted lysogeny-related genes.