Electrotransformation of Clostridium acetobutylicum ATCC 824 using high‐voltage radio frequency modulated square pulses

• Published in: Journal of applied microbiology Vol. 88; no. 2; pp. 220 - 227
• Main Authors: Tyurin, M., Padda, R., Huang, K. ‐X., Wardwell, S., Caprette, D., Bennett, G. N.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01-02-2000; Blackwell Science
• Subjects: Biological and medical sciences; Biotechnology; Clostridium - genetics; Clostridium - growth & development; Clostridium - ultrastructure; Clostridium acetobutylicum; Culture Media; Electric Stimulation - instrumentation; Electric Stimulation - methods; Electroporation - methods; Fundamental and applied biological sciences. Psychology; Gene transfer; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Microscopy, Electron; Plasmids - genetics; Synthetic digonucleotides and genes. Sequencing; Transformation, Bacterial; Clostridium acetobutylicum; Genetic transformation; Clostridiaceae; Clostridiales; Bacteria; Electroporation; Optimization; Genetic transfer
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1046/j.1365-2672.2000.00032.x

Molecular biological improvement of industrial solventogenic clostridia could be enhanced by a higher efficiency of electrotransformation. In this research, we used a new approach to determine the frequency spontaneously generated by Clostridium acetobutylicum ATCC 824 cells during the application of a square high‐voltage pulse. Once the frequency of 100 kHz was determined we transformed clostridial cells with pSOS84 plasmid DNA using radio‐frequency modulated high‐voltage square pulses (electric field strength 12 kVcm−1; pulse duration 22·5 ms; frequency of pulse modulation 100 kHz) to reach an efficiency exceeding 106 transformants μg−1 of plasmid DNA. We propose a possible role for cellular membrane structures in affecting the transformation yield.