Song S, Zhang T, Qi W, Zhao W, Xu B, Liu J
Department of Internal Medicine, Second Teaching Hospital, Tianjin Medical College, China.
Chin J Biotechnol. 1993;9(3):197-201.
Electroporation has been widely applied in molecular biology in recent years. In this study, successful transformation of plasmid DNA and transfection of phage DNA into E. coli were described by using intense electrical field of exponential decay waveform (electroporation) generated by a Gene Pulsar LN-101 (made in Tianjin, Tianjin Institute of Technology and Nankai University). We have obtained 10(9)-10(10) transformants/micrograms with strain DH5 alpha and plasmid pUC18, by a single voltage pulse at 1.0-2.5 kV (initial electric field strength = 2.8-16 kV/cm) with 5-20 microF capacitor. The efficiency of electroporation depends on various parameters: the electric field strength, capacitance, the pulse length (RC time constant), etc. The frequency of transformation was a linear function of the DNA concentration, as well as the density of recipient cell. The incubation time of before or after electroporation has no significant effect on transformation. The high transfection efficiency was also achieved with strain JM109 and M13mp19RF by electroporation. The method was easier, more time-saving and more efficient than Ca(2+)-dependent transformation (transfection) method. The results obtained by Gene-Pulsar LN-101 was similar to that by the Bio-Rad Gene-Pulsar apparatus.
近年来，电穿孔技术在分子生物学中得到了广泛应用。在本研究中，描述了使用由天津理工大学和南开大学制造的Gene Pulsar LN - 101产生的指数衰减波形强电场（电穿孔），将质粒DNA成功转化以及将噬菌体DNA转染到大肠杆菌中的过程。我们使用5 - 20微法的电容，在1.0 - 2.5 kV（初始电场强度 = 2.8 - 16 kV/cm）下施加单个电压脉冲，用菌株DH5α和质粒pUC18获得了10⁹ - 10¹⁰个转化子/微克。电穿孔效率取决于各种参数：电场强度、电容、脉冲长度（RC时间常数）等。转化频率是DNA浓度以及受体细胞密度的线性函数。电穿孔前后的孵育时间对转化没有显著影响。通过电穿孔，菌株JM109和M13mp19RF也实现了高转染效率。该方法比依赖Ca²⁺的转化（转染）方法更简便、更省时且更高效。Gene - Pulsar LN - 101获得的结果与Bio - Rad Gene - Pulsar仪器的结果相似。
