High osmotic stress improves electro-transformation efficiency of fission yeast.
A preincubation of fission yeast cells with hyperosmotic solution improved the electro-transformation efficiency. The efficiency increased approximately five-fold when the cells were preincubated with 2.0 M sorbitol and 1.5 M NaCl at 30 degrees C for 60 min before an applied high electric pulse. Losses in the efficiency of the cells after hyperosmotic stress above 2.5 M sorbitol and 2.0 M NaCl were directly related to the marked reduction of viability. The efficiency at 2.0 M sorbitol gradually increased until 60 min of the preincubation period, but longer exposure resulted in a gradual decrease. On the other hand, when the cells of the osmotic-sensitive mutant were preincubated with isosmotic solution of 0.5 M sorbitol, the efficiency was also dramatically increased by approximately 15-fold. These improvements in efficiency were observed in sublethal conditions of osmotic stress regardless of osmoticums and strains.
Suga
Minoru
M
Department of Materials and Biosystem Engineering, Toyama University, 3190 Gofuku Toyama-City, Toyama 930-8555, Japan. suga@eng.toyama-u.ac.jp
Kusanagi
Isamu
I
Hatakeyama
Toyomasa
T
D015169:Colony Count, Microbial; D018274:Electroporation / Q000379:methods*; D009994:Osmolar Concentration; D009997:Osmotic Pressure; D012568:Schizosaccharomyces / Q000235:genetics / Q000502:physiology*; D012965:Sodium Chloride / Q000494:pharmacology; D013012:Sorbitol / Q000494:pharmacology; D014170:Transformation, Genetic
D012965:Sodium Chloride; D013012:Sorbitol