关键词: 大肠埃希菌TG1, 电穿孔转化, 转化效率

Abstract: To build up a stable protocol of electroporation transformation method with high efficiency and improve storage capacity of genomie library,  we investigated the factors affecting the transformation efficiency, including culture temperature, growth state, voltage, the density and volume of competent cells, the amount of the exogenous gene and the volume of glycerol/mannitol buffer solution, which may contribute to construct highly diverse library. Results showed that when cells growing under 16℃ and harvested in mid-exponential phase (OD600 of 0.5), using glycerol/mannitol density gradient centrifugation techniques and adjusting the cell density up to 1×1011/mL, the electroporation voltage of 14.25 kV/cm, the highest transformation efficiency could reach to 9×109 CFU/μg DNA. The results showed that the optimal conditions for electroporation transformation would provide an important access for high-complexity library construction.

Key words: Escherichia coli TG1, electroporation, transformation efficiency