Development of the First Shuttle Vector System and Optimization of Transformation in Selenomonas sputigena

Area of Science:

• Microbiology
• Genetics
• Molecular Biology

Background:

• Selenomonas sputigena is an anaerobic bacterium implicated in periodontal disease and severe asthma.
• Lack of genetic tools hinders understanding of S. sputigena's molecular mechanisms and therapeutic potential.

Purpose of the Study:

• To develop the first genetic modification system for Selenomonas sputigena.
• To establish a standardized genetic tool for the Selenomonas genus.
• To enable investigation into S. sputigena's pathogenic and protective roles.

Main Methods:

• Construction of a shuttle vector using the Rep191 replication protein from Selenomonas ruminantium.
• Systematic optimization of transformation conditions (DNA quantity, growth phase, permeabilization, recovery time).
• Validation through transformation of multiple Selenomonas species and stable GFP expression.

Main Results:

• Achieved a 22.5-fold improvement in transformation efficiency for S. sputigena ATCC 35185.
• Established the first standardized genetic modification system applicable to multiple Selenomonas species.
• Demonstrated stable ectopic protein (GFP) expression in S. sputigena for the first time.

Conclusions:

• The developed genetic system provides essential tools for studying S. sputigena.
• This breakthrough facilitates research into the bacterium's dual role in human health.
• Potential for accelerating the development of microbiome-based treatments for respiratory and oral diseases.