ArticleFrontiers in microbiology2015
Construction of a Super-Competent Bacillus subtilis 168 Using the P mtlA -comKS Inducible Cassette.
Article in Frontiers in microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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40 citing papers in PubMed.
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- Metabolic Profile of the Genome-ReducedACS synthetic biology · 2024Article
- Biocomposite thermoplastic polyurethanes containing evolved bacterial spores as living fillers to facilitate polymer disintegration.Nature communications · 2024Article
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- Let There Be Light: Genome Reduction EnablesACS synthetic biology · 2023Article
- Development of a Tetracycline-Inducible System for Conditional Gene Expression in Lactococcus lactis and Streptococcus thermophilus.Microbiology spectrum · 2023Article
- Adaptive laboratory evolution ofMetabolic engineering communications · 2023Article
- Deletion of Rap-phosphatases for quorum sensing control in Bacillus and its effect on surfactin production.AMB Express · 2023Article
- Programming bacteria for multiplexed DNA detection.Nature communications · 2023Article
- Bacillus subtilis KM0 Impacts gut Microbiota Profile and Transcription of Genes Related to Transcellular Transport in Zebrafish (Danio rerio).Current microbiology · 2023Article
- CRISPR-dCas12a-mediated genetic circuit cascades for multiplexed pathway optimization.Nature chemical biology · 2023Article
- Membrane composition and organization of Bacillus subtilis 168 and its genome-reduced derivative miniBacillus PG10.Microbial biotechnology · 2022Article
- Surfactin Shows Relatively Low Antimicrobial Activity againstMicroorganisms · 2022Article
- Cross-Genus "Boot-Up" of Synthetic Bacteriophage in Staphylococcus aureus by Using a New and Efficient DNA Transformation Method.Applied and environmental microbiology · 2022Article
- Redirected Stress Responses in a Genome-Minimized 'midimSystems · 2021Article
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3 authors.
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Abstract
Competence is a physiological state that enables Bacillus subtilis 168 to take up and internalize extracellular DNA. In practice, only a small subpopulation of B. subtilis 168 cells becomes competent when they enter stationary phase. In this study, we developed a new transformation method to improve the transformation efficiency of B. subtilis 168, specially in rich media. At first, different competence genes, namely comK, comS, and dprA, were alone or together integrated into the chromosome of B. subtilis 168 under control of mannitol-inducible P mtlA promoter. Overexpression of both comK and comS increased the transformation efficiency of B. subtilis REG19 with plasmid DNA by 6.7-fold compared to the wild type strain 168. This transformation efficiency reached its maximal level after 1.5 h of induction by mannitol. Besides, transformability of the REG19 cells was saturated in the presence of 100 ng dimeric plasmid or 3000 ng chromosomal DNA. Studying the influence of global regulators on the development of competence pointed out that important competence development factors, such as Spo0A, ComQXPA, and DegU, could be removed in REG19. On the other hand, efficient REG19 transformation remained highly dependent on the original copies of comK and comS regardless of the presence of P mtlA -comKS. Finally, novel plasmid-free strategies were used for transformation of REG19 based on Gibson assembly.
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