Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider P H
EMBO J. 1982;1(7):841-5. doi: 10.1002/j.1460-2075.1982.tb01257.x.
Electric impulses (8 kV/cm, 5 microseconds) were found to increase greatly the uptake of DNA into cells. When linear or circular plasmid DNA containing the herpes simplex thymidine kinase (TK) gene is added to a suspension of mouse L cells deficient in the TK gene and the cells are then exposed to electric fields, stable transformants are formed that survive in the HAT selection medium. At 20 degrees C after the application of three successive electric impulses followed by 10 min to allow DNA entry there result 95 (+/- 3) transformants per 10(6) cells and per 1.2 micrograms DNA. Compared with biochemical techniques, the electric field method of gene transfer is very simple, easily applicable, and very efficient. Because the mechanism of DNA transport through cell membranes is not known, a simple physical model for the enhanced DNA penetration into cells in high electric fields is proposed. According to this ' electroporation model' the interaction of the external electric field with the lipid dipoles of a pore configuration induces and stabilizes the permeation sites and thus enhances cross membrane transport.
研究发现，电脉冲（8千伏/厘米，5微秒）能极大地增加细胞对DNA的摄取。当将含有单纯疱疹胸苷激酶（TK）基因的线性或环状质粒DNA添加到缺乏TK基因的小鼠L细胞悬液中，然后使细胞暴露于电场时，会形成稳定的转化体，这些转化体能够在HAT选择培养基中存活。在20摄氏度下，施加三个连续的电脉冲，随后经过10分钟以使DNA进入细胞，每10⁶个细胞和每1.2微克DNA会产生95（±3）个转化体。与生化技术相比，电场基因转移方法非常简单、易于应用且效率极高。由于DNA通过细胞膜的运输机制尚不清楚，因此提出了一个简单的物理模型来解释在高电场中DNA增强进入细胞的现象。根据这个“电穿孔模型”，外部电场与孔构型的脂质偶极相互作用会诱导并稳定渗透位点，从而增强跨膜运输。
