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| 克雷伯氏肺炎杆菌电击转化条件的优化 | |
| 引用本文： | 魏东,王敏,庄文超,史吉平,郝健坤.克雷伯氏肺炎杆菌电击转化条件的优化[J].中国酿造,2012,31(4):18-21. |
| 作者姓名： | 魏东 王敏 庄文超 史吉平 郝健坤 |
| 作者单位： | 1. 工业发酵微生物教育部重点实验室(天津科技大学)生物工程学院天津科技大学天津300457;中国科学院上海高等研究院,上海201210 2. 工业发酵微生物教育部重点实验室(天津科技大学)生物工程学院天津科技大学天津300457 3. 中国科学院上海高等研究院,上海,201210 |
| 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。 |
| 关 键 词： | 克雷伯氏肺炎杆菌 电击转化 效率 |
Optimization the electroporation parameters of Klebsiella pneumoniae |
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| WEI Dong , WANG Min , ZHUANG Wenchao , SHI Jiping , HAO Jian.Optimization the electroporation parameters of Klebsiella pneumoniae[J].China Brewing,2012,31(4):18-21. | |
| Authors: | WEI Dong WANG Min ZHUANG Wenchao SHI Jiping HAO Jian |
| Affiliation: | 1.Key Laboratory of Industrial Fermentation Microbiology(Tianjin University of Science ＆ Technology),Ministry of Education, College of Biotechnology,Tianjin University of Science and Technology,Tianjin 300457,China; 2.Shanghai Advanced Research Institute,Chinese Academy of Sciences,Shanghai 201210,China) |
| Abstract: | 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. |
| Keywords: | Klebsiella pneumonia electroporation efficiency |
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