Fast kinetic studies of plasmid DNA transfer in intact yeast cells mediated by electropulsation.
Intact yeast cell Electrotransformation process was investigated. It is a two step process. The plasmid must be pre-mixed and present in contact with the cells during the pulse. During the millisecond field pulse, plasmid DNA is associated to the envelope. It therefore crosses the membrane by a process which lasts several seconds as shown by its sensitivity to a post pulse addition of DNase. Electrotransformation is not supported by an electrophoretic transfer due to the external field nor by a free diffusion across the electropermeabilized envelope. DNA is first bound during the field pulse and then is transferred by a still unknown active process due to cell metabolism.
Ganeva
V
V
Dept 3, Glycoconjugués et Biomembranes, LPTF CNRS, Toulouse, France.
Galutzov
B
B
Teissie
J
J
D001692:Biological Transport; D003851:Deoxyribonucleases; D004558:Electric Stimulation / Q000379:methods; D018274:Electroporation / Q000379:methods; D007700:Kinetics; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014162:Transfection / Q000379:methods*
D003851:Deoxyribonucleases