Transposon insertion in Rothia dentocariosa

Area of Science:

• Microbiology
• Genetics
• Oral Health

Background:

• Rothia spp. are significant oral bacteria, with Rothia dentocariosa being highly prevalent.
• Limited genetic-level studies exist for Rothia spp., hindering a deeper understanding of their oral health impact.

Purpose of the Study:

• Establish a genetic modification platform for Rothia dentocariosa.
• Enable future genetic investigations into the role of R. dentocariosa in the oral microbiome.

Main Methods:

• Isolation and identification of Rothia spp. from saliva samples using PCR and 16S rRNA sequencing.
• Transformation of R. dentocariosa strains with plasmid pJRD215 to identify a highly transformable strain (LX16).
• Application of transposon insertion mutagenesis and screening for resistant mutants to confirm genetic modification feasibility.

Main Results:

• Identification of Rothia dentocariosa LX16, a strain with high transformation efficiency.
• Successful induction of transposon insertion mutagenesis in R. dentocariosa LX16.
• Confirmation of transposon insertion sites using arbitrary primed PCR, gene-specific PCR, and Sanger sequencing.

Conclusions:

• This study reports the first successful genetic modification of Rothia dentocariosa.
• The established platform facilitates genetic analysis of R. dentocariosa.
• Genetic insights into R. dentocariosa will enhance understanding of its function within the oral microbiome.