应用电激法在大肠杆菌中导入外源性DNA  

Electroporation of Foreigh DNA into Escherichia coli

作　　者：宋诗铎[1] 张同海[1] 祁伟[1] 赵为诚[1] 徐宝强[2] 刘建民[2] 

机构地区：[1]天津医学院第二附属医院,天津 300211 [2]天津理工学院,天津 300191

出　　处：《生物工程学报》1993年第3期237-240,共4页Chinese Journal of Biotechnology

摘　　要：本文报道用国产基因导入脉冲仪(LN-101型),采用高压脉冲电场,将质粒DNA和噬菌体DNA成功地转化或转导入大肠杆菌。该电场单次电压脉冲范围1.0kV—2.5kV(初电场强度2.8kV/cm—16kV/cm)、电容范围5—20μF,用质粒pUC18和受体菌株DH5α,获得10~9—10^(10)转化子/μgDNA。电场强度、电容及脉冲时间等不同的因素影响转化效率。转化率与DNA的强度呈线性关系,与受体细胞密度成正比。DNA和受体菌混合物在电激前、后的孵育时间,对转化效率影响不明显。用噬菌体M13mp 19 RF及受体菌株JM109,同样可获得较高的转化效率。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 generated by a Gene Pulser LN-101. We have obtained 10~9—10^(10) transformants/μg DNA with strain DH 5 a, and plasmid pUC 18, by a single voltage pulse at 1.0—2.5 kV with 5—20μF capacitor. The efficiency of electroporation depends on various parameters: the electric field strength, capacitance, the pulse length etc. The frequency of transformation was a linear function of the DNA concentration, same as the density of recipient cell. The effect of time of preand post-shoke incubation of cell with DNA was insignificient. The high efficiency of transfection also was achieved with strain JM 109 and M 13 mp 19 RF by electroporation.

关 键 词：DNA 电激法 转导 大肠杆菌 

分 类 号：Q523.03[生物学—生物化学]