ArticleFrontiers in microbiology2016
Overcoming the Barrier of Low Efficiency during Genetic Transformation of Streptococcus mitis.
Article in Frontiers in microbiology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 39 citations in OpenAlex.
- RggmBio · 2026Article
- Genetic and functional characterization of the natural transformation system inMicrobiology (Reading, England) · 2026Article
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- The Quorum Sensing-Controlled Competence Regulon Drives HACS infectious diseases · 2025Article
- TrkA ofMicrobiology (Reading, England) · 2025Article
- Deciphering the Regulatory Role and Molecular Interactions That Drive the Competence Regulon Quorum Sensing Circuitry inBiochemistry · 2025Article
- Elucidating the Role of the Competence Regulon Quorum Sensing Circuitry inACS chemical biology · 2025Article
- Inhibition of pneumococcal growth and biofilm formation by human isolates ofApplied and environmental microbiology · 2025Article
- Article
- Biosynthesis of mitis group streptococcal glycolipids and their roles in physiology and antibiotic susceptibility.bioRxiv : the preprint server for biology · 2024Article
- Genetic modification ofmSphere · 2024Article
- Illuminating the oral microbiome and its host interactions: tools and approaches for molecular microbiology studies.FEMS microbiology reviews · 2023Review
- Investigating theMicrobiology (Reading, England) · 2022Article
- Elucidating the Role and Structure-Activity Relationships of theACS chemical biology · 2021Article
- Streptococcus pneumoniae, S. mitis, and S. oralis Produce a Phosphatidylglycerol-Dependent,mSphere · 2021Article
- Cyclic Di-adenosine Monophosphate Regulates Metabolism and Growth in the Oral CommensalMicroorganisms · 2020Article
- Biological Evaluation of Native Streptococcal Competence Stimulating Peptides Reveal Potential Crosstalk BetweenRSC chemical biology · 2020Article
- Phosphatidylcholine Biosynthesis in Mitis Group Streptococci via Host Metabolite Scavenging.Journal of bacteriology · 2019Article
- Characterization of a Signaling System in Streptococcus mitis That Mediates Interspecies Communication with Streptococcus pneumoniae.Applied and environmental microbiology · 2019Article
- Competence inFrontiers in cellular and infection microbiology · 2019Review
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
objectiveStreptococcus mitis is a predominant oral colonizer, but difficulties in genetic manipulation of this species have hampered our understanding of the mechanisms it uses for colonization of oral surfaces. The aim of this study was to reveal optimal conditions for natural genetic transformation in S. mitis and illustrate its application in direct genome editing.
methodsLuciferase reporter assays were used to assess gene expression of the alternative sigma factor (σ(X)) in combination with natural transformation experiments to evaluate the efficiency by which S. mitis activates the competence system and incorporates exogenous DNA. Optimal amounts and sources of donor DNA (chromosomal, amplicon, or replicative plasmid), concentrations of synthetic competence-stimulating peptide, and transformation media were assessed.
resultsA semi-defined medium showed much improved results for response to the competence stimulating peptide when compared to rich media. The use of a donor amplicon with large homology flanking regions also provided higher transformation rates. Overall, an increase of transformation efficiencies from 0.001% or less to over 30% was achieved with the developed protocol. We further describe the construction of a markerless mutant based on this high efficiency strategy.
conclusionWe optimized competence development in S. mitis, by use of semi-defined medium and appropriate concentrations of synthetic competence factor. Combined with the use of a large amplicon of donor DNA, this method allowed easy and direct editing of the S. mitis genome, broadening the spectrum of possible downstream applications of natural transformation in this species.
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