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| 制备高效大肠杆菌电转化感受态细胞和电转化条件的研究 | |
| 引用本文： | 朱森康,黄磊,李燕飞,钟卫鸿,徐志南.制备高效大肠杆菌电转化感受态细胞和电转化条件的研究[J].生物技术通报,2011(10). |
| 作者姓名： | 朱森康 黄磊 李燕飞 钟卫鸿 徐志南 |
| 作者单位： | 1. 浙江工业大学生物与环境工程学院,杭州 310032;浙江大学化学工程与生物工程学系,杭州310027 2. 浙江大学化学工程与生物工程学系,杭州,310027 3. 浙江大学化学工程与生物工程学系,杭州310027;浙江工业大学药学院,杭州310032 4. 浙江工业大学生物与环境工程学院,杭州,310032 |
| 基金项目： | 国家自然科学基金项目(21006088); 浙江省自然科学基金项目(Y4100344) |
| 摘 要： | 旨在建立一种高效大肠杆菌电转化感受态细胞的制备方法,研究了摇瓶装液量,菌体生长阶段,转化电场强度,转化后复苏时间以及感受态细胞存放时间等对转化效率的影响.结果表明,基液量为400 mL/2L,菌体在OD600值为0.452时收集所制备的感受态细胞,在电场强度12.5 kV/cm条件下电击5 ms,转化后复苏时间2h,转化效率可达到1010CFU/μg DNA.此条件下制备的感受态细胞转化效率高,质量稳定,重复性好. |
| 关 键 词： | 大肠杆菌 电转化 感受态细胞 转化效率 |
Preparation of Efficient Electrotransformation Competent Cells of Escherichia coli and Condition of Electrotransformation |
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| Zhu Senkang,Huang Lei,Li Yanfei,Zhong Weihong,Xu Zhinan.Preparation of Efficient Electrotransformation Competent Cells of Escherichia coli and Condition of Electrotransformation[J].Biotechnology Bulletin,2011(10). | |
| Authors: | Zhu Senkang Huang Lei Li Yanfei Zhong Weihong Xu Zhinan |
| Institution: | Zhu Senkang1,2 Huang Lei2 Li Yanfei2,3 Zhong Weihong1 Xu Zhinan2(1College of Biological and Environmental Engineering,Zhejiang University of Technology,Hangzhou 310032,2 Department Chemical and Biological Engineering,Zhejiang University,Hangzhou 310027,3College of Pharmaceutical Science,Hangzhou 310032) |
| Abstract: | The aim of this study was to optimize the preparation of E.coli electrotransformation competent cells.In the present work,the effects of different prepare conditions were systematically investigated to improve the transformation efficiency of the competent cells.The optimal conditions were determined as follows: cultivation in 400 ml/2 L LB medium,collection of the cells at OD600 of 0.452,electrotransformation at 12.5 kV/cm for 5 ms,posttransformation culture for 2 h.Under these conditions,the transformatio... |
| Keywords: | Escherichia coli Electrotransformation Competent cells Transformation efficiency |
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