摘要
针对克雷伯氏肺炎杆菌(Klebsiella pneumoniae)TUAC01的电击转化方法进行了研究。主要考察了培养基EDTA的浓度、细胞的生长状态、感受态细胞的密度、电击电压、质粒浓度等参数对转化效率的影响,建立了一种适合K.pneumoniae TUAC01的电击转化方法。结果:K.pneumoniae TUAC01培养基中添加EDTA至终浓度0.7mmol/L;当细胞生长至OD600为0.7时收集菌体,制作感受态细胞;制作的感受态细胞浓度OD600大于30时,使用2mm的电转杯在2kV的电压下转化,转化效率达到最大值,达到8.58×105cfu/μg DNA。
The electroporation parameters of Klebsiella pneumoniae TUAC01 were optimized based on the electroporation efficiency.The effect of operate parameters on the electroporation efficiency was investigated,those operated parameters included pulser voltage,cell growth stage,final electro-competent cell density,EDTA level in medium and plasmid concentration.It was shown that,adding EDTA to a final concentration of 0.7mmol/L in medium,collect cell at the stage of cell density reached OD600 0.7,concentrated the electro-competent cell to a final density of larger than OD600 30 and pulser voltage selected 2kV used 2mm electroporate chamber.Under this condition,the electroporation efficiency reached to 8.58×105cfu/μg DNA transformants.
出处
《中国酿造》
CAS
2012年第4期18-21,共4页
China Brewing
基金
国家自然科学基金(20906076)
