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 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 microg-1 of plasmid DNA. We propose a possible role for cellular membrane structures in affecting the transformation yield.
Tyurin
M
M
Department of Biochemistry and Cell Biology, and Department of Ecology and Evolutionary Biology, Rice University, Houston, TX 77251-1892, USA.
Padda
R
R
Huang
K X
KX
Wardwell
S
S
Caprette
D
D
Bennett
G N
GN
D003013:Clostridium / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D003470:Culture Media; D004558:Electric Stimulation / Q000295:instrumentation / Q000379:methods; D018274:Electroporation / Q000379:methods*; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics; D014169:Transformation, Bacterial
D003470:Culture Media