Transformation of Bacillus amyloliquefaciens by electroporation

• Published in: FEMS microbiology letters 1989-10, Vol.61 (1‐2), p.165-170
• Main Author: VEHMAANPERA, J
• Format: Article
• Language: English
• Subjects: Bacillus - genetics; Bacillus - growth & development; Bacillus subtilis; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; DNA; DNA, Bacterial - isolation & purification; Electrochemistry; Electropermeabilization; electroporation; Electro‐transformation; Fundamental and applied biological sciences. Psychology; Microbiology; pC194; pE194; Plasmids; pUB110; transformation; Transformation, Bacterial
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.1989.tb03572.x

A method for transformation of whole Bacillus amyloliquefaciens cells by electroporation was developed. The procedure is as efficient as the protoplast transformation method, resulting in up to 105 transformants/μg plasmid DNA, but requires less effort and time. Cells for electroporation were grown to late exponential phase in a rich medium supplemented with 0.25 M sucrose, washed with and resuspended in 0.25 M sucrose, 1 mM HEPES, 1 mM MgCl2, 10% (v/v) glycerol, pH 7.0, at 3–5 × 1010 cells/ml for storage at ‐80%°C. The highest transformation frequency was obtained at 7.5 kV/cm with a 25 μF capacitor. The transformation efficiency increased linearly with DNA concentration at least over the range 10 ng‐12.5 μg/ml. Transformations with ligated DNA and of industrial strains were also successful. In addition, B. subtilis cells treated as above could be transformed by electroporation, resulting in 104 transformants/μg DNA at 12.5 kV/cm.