电转化条件对大肠杆菌TG1转化效率的影响

Impact of Electroporation on Transformation Efficiency of Escherichia(E.) coli TG1

研究探索不同条件对TG1大肠杆菌转化率的影响,以探索最适合电转化条件。研究通过对细菌生长曲线绘制,改变不同生长阶段、感受态细胞浓度、转化后复苏液以及复苏时间、电转化参数、质粒浓度、抗生素浓度等影响电转化率的条件,探索最适合的电转化条件。结果表明,TG1大肠杆菌37℃、170 rpm/min震荡活化培养时间3 h后,以电压1 800 V、电容25μF、电阻200Ω进行电击转化,以LB液体对电击细胞复苏45 min,为最佳转化条件。该结果具有可重复性,为后续建立基因文库奠定良好基础。

This paper explores the optimal condition of electroporation with an analysis on the transformation efficiency of Escherichia( E.) coli TG1 under the different conditions. The optimal condition of electroporation was explored with a study on the conditions,which can impact on the transformation efficiency,through mapping of the growth curve of TG1 by changing the incubation period,competent cell concentration,recovery solution and period,parameters of electric shock,plasmid concentration,and antibiotic concentration. The results show that the optimal condition of electrotransformation for TG1 is as followed,cultured at 37 ℃ with 170 rpm/min for 3 h,shocked in the condition( 1 800 V,25 μF,200 Ω),recovered with LB medium for 45 min,contained 1. 4 × 10~7 of competent cell. The results are repeatable,and lay the foundation for subsequent establishment of the library.