Effects of DNA Topology on Transformation Efficiency of<i>Bacillus subtilis</i>ISW1214 by Electroporation

Ohse, Morimasa; Kawade, Koji; Kusaoke, Hideo

doi:10.1271/bbb.61.1019

Cited by 17 publications

(8 citation statements)

References 6 publications

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“…Electroporation is usually used for transformation of B. subtilis (Kusaoke et al 1989;Ohse et al 1997). The transformation efficiency of B. subtilis varies widely among strains.…”

Section: Discussionmentioning

confidence: 99%

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Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis

Zhang

et al. 2012

Letters in Applied Microbiology

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Aims: To optimize the transformation conditions and improve the transformation efficiency of Bacillus subtilis WB800 and DB104. Methods and Results: Trehalose, which could decrease the damage of electric shock to the cells, was added to the electroporation medium containing sorbitol and mannitol. The factors affecting the transformation efficiency, such as the growth phase of bacteria, cell concentration, electric field strength and plasmid variety, were examined and improved. The new method increased the transformation efficiency of B. subtilis by nearly 100‐fold compared with the conventional one. Conclusions: With the optimized method, the transformation efficiency came up to 3·64 × 105 transformants μg−1 DNA for WB800, and 2·10 × 105 transformants μg−1 DNA for DB104. Significance and Impact of the Study: This improvement in transformation efficiency will be largely attributed to the research of expression of exogenous genes in B. subtilis, gene library construction for directed evolution and transformation of wild‐type B. subtilis strains.

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“…Electroporation is usually used for transformation of B. subtilis (Kusaoke et al 1989;Ohse et al 1997). The transformation efficiency of B. subtilis varies widely among strains.…”

Section: Discussionmentioning

confidence: 99%

“…Electroporation is usually used for transformation of B. subtilis (Kusaoke et al. 1989; Ohse et al. 1997).…”

Section: Discussionmentioning

confidence: 99%

Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis

Zhang

et al. 2012

Letters in Applied Microbiology

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“…Size, topology, and sequence of DNA have been mentioned in previous work investigating properties of DNA that affect natural competence, but no quantitative results are available to correlate the size of DNA with transformation efficiency . Given that most commonly used double‐crossover integration vectors are about 6–10 kb, we added non‐essential sequences to our assembled linear DNA to make it longer.…”

Section: Resultsmentioning

confidence: 99%

“…With the elucidation of the regulatory network of natural competence , the gene encoding the master regulator ComK was placed under the control of inducible promoters to transiently switch on the natural competence and significantly increased the transformation efficiency . DNA uptake specificity was investigated to uncover the relationship among DNA topology, sequence and transformation efficiency . Electroporation and protoplast fusion were also used for genetic transformation in B. subtilis , particularly for strains without natural competence .…”

Section: Introductionmentioning

confidence: 99%

Fast genome editing in Bacillus subtilis

Drufva

2019

Engineering in Life Sciences

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Bacillus subtilis is a model organism for Gram‐positive bacteria and widely used in the study of cellular functions and processes including protein secretion, sporulation, and signal transduction. It is also an important industrial host for the production of proteins and chemicals. Generally, genome editing of B. subtilis often needs the construction of integration vectors in Escherichia coli, linearizing the constructed plasmids, and subsequent transformation of the linear deoxyribonucleic acid via natural competence or electroporation. In this work, we examined the feasibility to directly transform and integrate B. subtilis using linear deoxyribonucleic acid from Gibson assembly without the need for cloning in E. coli. Linear deoxyribonucleic acid of 8–10 kb showed the highest transformation efficiency which was similar to that of using linearized plasmids constructed in E. coli. This method shortens the overall process from 1 week to 1 day and allows the integration of multiple genes in one step, providing a simple and fast method for genome editing in B. subtilis.

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“…Cell electrotransformation with circular supercoiled DNA gives the maximum efficiency, higher than for the circular relaxed DNA (Ohse, et al, 1997). The transformation efficiency for the linearized DNA is extremely low or non-existant , Kimoto and Taketo, 1996, Ohse, et al, 1997.…”

Section: Effects Of Size and Molecular Form Of Dna And Plasmid Concenmentioning

confidence: 97%

General Principles of Bacteria Electrotransformation: Key Steps

Eynard

Teissié

2000

Electrotransformation of Bacteria

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Effects of DNA Topology on Transformation Efficiency ofBacillus subtilisISW1214 by Electroporation

Cited by 17 publications

References 6 publications

Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis

Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis

Fast genome editing in Bacillus subtilis

General Principles of Bacteria Electrotransformation: Key Steps

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