title: Genome sequencing of a genetically tractable derivative of Fusobacterium nucleatum subsp. nucleatum ATCC 25586
design: Fusobacterium nucleatum is an anaerobic Gram-negative bacterium commonly found in the human oral cavity. Although present at low abundance in healthy subgingival plaque, it becomes highly enriched in periodontal pockets during periodontal disease. Beyond oral infections, increasing evidence links F. nucleatum to several systemic conditions, including colorectal cancer, adverse pregnancy outcomes, inflammatory bowel disease, and rheumatoid arthritis. F. nucleatum subsp. nucleatum ATCC 25586, originally isolated from a cervicofacial lesion, has been widely used as a reference strain for studies of fusobacterial physiology and pathogenesis. However, the ATCC 25586 strain obtained directly from ATCC has been reported to be genetically intractable in several laboratories, limiting its use in genetic studies.Here we report the whole-genome sequence of a genetically tractable variant of F. nucleatum ATCC 25586 (we named ATCC25586wu) obtained from Dr. Amanda L. Lewis (University of California San Diego), originally purchased from ATCC. The strain was revived from a minus 80 C glycerol stock by direct inoculation into 8 mL of trypticase soy broth supplemented with 1 percent peptone and incubated anaerobically at 37 C for 14 hours. Cells were harvested by centrifugation, and genomic DNA was extracted using the ZymoBIOMICS DNA Miniprep Kit (Zymo Research, D4300) following the manufacturer's protocol. DNA concentration was determined using a Qubit 4 fluorometer (Thermo Fisher Scientific, Q33327). Sequencing was performed by SeqCenter (Pittsburgh, PA) using both Illumina and Oxford Nanopore Technologies platforms. The genome comprises a 2180128 bp chromosome.