Oral

[O2-E-AM2-05] Development of a novel gene editing technologies using Rothia spp.

〇Boang Liu¹, Chiho Mashimo², Takayuki Nambu², Hugo Maruyama², Toshinori Okinaga² (1. Dept Bacteriol, Osaka Dent Univ Grad Sch Dent, 2. Dept Bacteriol, Osaka Dent Univ)

Keywords:Rothia spp.、transposon mutagenesis、Nitrate reduction genes

It is known that Rothia spp. are abundant in a healthy oral cavity and speculated that their nitrate-reducing ability might contribute to maintain oral health. Although detailed researches are needed, there is no method available for analyzing characteristics of Rothia spp. at the genetic level. To overcome the aforementioned issue, we tried to isolate genetically tractable Rothia spp. from oral cavity. The strain with highest transformation efficiency was selected and named Rothia dentocariosa LX16. We performed transposon mutagenesis of R. dentocariosa LX16 using EZ-Tn5^TM Tnp Transposome ^TM (Epicentre; WI, USA) and constructed genome-wild mutant library. The transposon insertion sites were confirmed by arbitrary primed polymerase chain reaction (AP-PCR) and nanopore sequencing. We selected nitrate reduction-deficient clones and confirmed candidate genes, by screening based on Griess reaction. Further developing genetic techniques could elucidate the role of Rothia spp. in the oral cavity.