ArticleInternational journal of molecular sciences2015
Development of an efficient electroporation method for iturin A-producing Bacillus subtilis ZK.
Article in International journal of molecular sciences, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed.
- The Effect ofMicroorganisms · 2026Article
- Integrated metagenomic-metabolomic insights into plant-microbe interactions mediated byApplied and environmental microbiology · 2026Article
- Development of a Tool for High-Efficiency, Markerless and Iterative Genome Editing in Shouchella clausii.Microbial biotechnology · 2026Article
- An efficient electrotransformation method for three Bacillus species.Applied microbiology and biotechnology · 2025Article
- High-Throughput Microfluidic Electroporation (HTME): A Scalable, 384-Well Platform for Multiplexed Cell Engineering.Bioengineering (Basel, Switzerland) · 2025Article
- Construction of an efficient electroporation transformation system promotes the application of Targetron in wild-typeApplied and environmental microbiology · 2025Article
- Development of genetic manipulation tools forFrontiers in microbiology · 2025Article
- Influence of PepF peptidase and sporulation on microcin J25 production inMicrobiology spectrum · 2024Article
- Calculating transmembrane voltage on the electric pulse-affected cancerous cell membrane: using molecular dynamics and finite element simulations.Journal of molecular modeling · 2024Article
- Cell Factory Engineering of UndomesticatedFrontiers in microbiology · 2022Article
- Implantation ofFrontiers in microbiology · 2022Article
- Surfactin secreted byFrontiers in plant science · 2022Article
- Factors and Conditions That Impact Electroporation of Clostridioides difficile Strains.mSphere · 2020Article
- Optimization and Comparison of Different Methods and Factors for Efficient Transformation ofAdvanced biomedical research · 2019Article
- Construction of a plasmid interspecific transfer system in Bacillus species with the counter-selectable marker mazF.Journal of industrial microbiology & biotechnology · 2018Article
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5 authors.
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Abstract
In order to efficiently introduce DNA into B. subtilis ZK, which produces iturin A at a high level, we optimized seven electroporation conditions and explored an efficient electroporation method. Using the optimal conditions, the electroporation efficiency was improved to 1.03 × 10(70 transformants/μg of DNA, an approximately 10,000-fold increase in electroporation efficiency. This efficiency is the highest electroporation efficiency for B. subtilis and enables the construction of a directed evolution library or the knockout of a gene in B. subtilis ZK for molecular genetics studies. In the optimization process, the combined effects of three types of wall-weakening agents were evaluated using a response surface methodology (RSM) design, which led to a two orders of magnitude increase in electroporation efficiency. To the best of our limited knowledge, this study provides the first demonstration of using an RSM design for optimization of the electroporation conditions for B. subtilis. To validate the electroporation efficiency, a case study was performed and a gene (rapC) was inactivated in B. subtilis ZK using a suicide plasmid pMUTIN4. Moreover, we found that the rapC mutants exhibited a marked decrease in iturin A production, suggesting that the rapC gene was closely related to the iturin A production.
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