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

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 o...

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Veröffentlicht in:Journal of applied microbiology 2000-02, Vol.88 (2), p.220-227
Hauptverfasser: Tyurin, M., Padda, R., Huang, K. ‐X., Wardwell, S., Caprette, D., Bennett, G. N.
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Sprache:eng
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Zusammenfassung: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.
ISSN:1364-5072
1365-2672
DOI:10.1046/j.1365-2672.2000.00032.x