title: Nanopore raw read
design: Fusobacterium nucleatum is a Gram-negative, spindle-shaped bacterium associated with various oral diseases and systemic infections . A key feature of this organism is its ability to physically interact with numerous oral bacterial species and to adhere to multiple types of mammalian cells. During early studies investigating the molecular basis of F. nucleatummediated cellcell interactions, two strains were commonly used: ATCC 25586 and PK1594. F. nucleatum represents a highly diverse group currently divided into four subspeciesnucleatum (FNN), vincentii (FNV), polymorphum (FNP), and animalis (FNA)which have recently been proposed to constitute distinct species. Based on gene, genomic, and biochemical evidence, ATCC 25586 has been classified as FNN. However, no genome sequence or classification data were available for strain PK1594 until now.Strain PK1594 was originally isolated from a human subgingival site and first reported in 1982 as Fusobacterium nucleatum strain VPI E2S-11A. In 1989, it was renamed PK1594 when deposited in the laboratory of the late Dr. Paul E. Kolenbrander (NIDCR, NIH). We obtained this strain from Dr. Robert J. Palmer (NIH) and have maintained it in our laboratory. The strain was revived from a 80C glycerol stock by direct inoculation into 8 mL of trypticase soy broth supplemented with 1% peptone and incubated anaerobically at 37C for 14 hours. Cells were harvested by centrifugation, and genomic DNA was extracted using the ZymoBIOMICS DNA Miniprep Kit (Zymo Research, D4300) following the manufacturers 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 (ONT) platforms. The genome comprises a 2,357,192-bp chromosome and a plasmid originally assembled as 20,441 bp but representing a circular molecule of approximately 10,221 bp. Gene analysis identified PK1594 as Fusobacterium hwasookii.