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Electrotransformation of Clostridium acetobutylicum ATCC 824 using high‐voltage radio frequency modulated square pulses
Author(s) -
Tyurin M.,
Padda R.,
Huang K. X.,
Wardwell S.,
Caprette D.,
Bennett G. N.
Publication year - 2000
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2000.00032.x
Subject(s) - electroporation , clostridia , plasmid , clostridium acetobutylicum , pulse (music) , voltage , transformation (genetics) , transformation efficiency , clostridium , biophysics , chemistry , microbiology and biotechnology , biology , dna , bacteria , biochemistry , physics , gene , genetics , butanol , quantum mechanics , ethanol , agrobacterium
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 10 6 transformants μg −1 of plasmid DNA. We propose a possible role for cellular membrane structures in affecting the transformation yield.