Influence of pulsing and postpulsing media tonicity on electrotransformation of intact yeast cells.
The maximal transformation yield of intact yeast cells in hypotonic medium was obtained by application of nine pulses with a duration of 990 microseconds at 2.5 kV cm-1. Pulsation at the same electrical parameters in isotonic solution did not lead to any transformation and the electropermeability decreased by 50%. The transfer of cells, 1 min after pulsation in hypotonic medium, into media with different tonicity led to an increase of the number of transformed cells, depending on the sorbitol concentration of up to 250 mM. Further augmentation of the tonicity of postpulse medium in the range 330-1000 mM provoked strong decrease of transformation. This effect was present even when cells were resuspended in isotonic medium 30 min after pulsation.
Ganeva
V
V
Department of Biophysics and Radiobiology, Sofia University, Bulgaria.
Galutzov
B
B
D004271:DNA, Fungal / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D005069:Evaluation Studies as Topic; D007038:Hypotonic Solutions; D009995:Osmosis; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013012:Sorbitol; D014170:Transformation, Genetic
D004271:DNA, Fungal; D007038:Hypotonic Solutions; D013012:Sorbitol