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노지윤,함영태,정미현 中央大學校 遺傳工學硏究所 1994 遺傳工學硏究論集 Vol.7 No.1
Electroportation, the reversible breakdown of cell membrane caused by a high-voltage discharge, is rapid, simple, and efficient method for introducing DNA into the cells. Electroporation was developed as the powerful method for the genetic work with eukaryotic cells. Very few reports have been published where this method has been developed for prokaryotic systems. Electroportation has one major advantage: It works well with cell lines that are refractive to other techniques, such as calcium phosphate-DNA coprecipitation for the particular cell lines under study. For yeast, the cell death rate increases with the increment of thd voltage. The death rate is 52% at 900 V/0.4 ㎝ and goes up to 84% at 1000 V/0.4 ㎝. For the high transformation efficiency the optimum voltage is around 900 V. The charge of the eletric resistance at the fixed voltage(2.0 kv/㎝) and capacitance(25 uF) leads to decrease abruptly cell survial. The optimum condition 25 uF and eledtric resistance 800 ohm. The condition with the pulses 20 msec and field strength 7 kv/㎝ gave the maximum transformation efficiency in E. coli. The post-pulse incubation is also in fluence on the transformation efficience.