Simple method for demonstrating small plasmid deoxyribonucleic acid molecules in oral streptococci

Area of Science:

• Microbiology
• Molecular Biology
• Genetics

Background:

• Oral streptococci are significant in dental caries and oral health.
• Plasmids play a vital role in bacterial adaptation and virulence.
• Efficient methods for plasmid detection are crucial for studying oral bacteria.

Purpose of the Study:

• To develop a rapid and simple procedure for small plasmid demonstration in oral streptococci.
• To enable visualization of plasmids less than 10 megadaltons.

Main Methods:

• Utilized logarithmic-phase, glycine-treated oral streptococci from small broth cultures (1.5 ml).
• Induced osmotic fragility and cell lysis using sodium dodecyl sulfate.
• Employed hydrodynamic shearing to fragment host chromosomal deoxyribonucleic acid.
• Analyzed cell lysates via low-voltage agarose gel electrophoresis.

Main Results:

• Successfully demonstrated small plasmids (<10 megadaltons) in oral streptococci.
• Observed plasmids migrating significantly faster than chromosomal deoxyribonucleic acid on gels.
• Ethidium bromide staining allowed for clear visualization of the detected plasmids.

Conclusions:

• The described method offers a rapid and straightforward approach for small plasmid detection in oral streptococci.
• This technique facilitates the study of plasmid-borne traits in these bacteria.
• The procedure is suitable for routine laboratory use in microbiology and molecular biology research.