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| 连续热裂解法大规模提取质粒DNA | |
| 引用本文： | 朱开春,张富春,肖冲,何忠淮,金华利,王宾. 连续热裂解法大规模提取质粒DNA[J]. 农业生物技术学报, 2006, 14(1): 51-54 |
| 作者姓名： | 朱开春 张富春 肖冲 何忠淮 金华利 王宾 |
| 作者单位： | 1. 新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐,830046 2. 中国农业大学生物学院农业生物技术国家重点实验室,北京,100094 3. 新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐,830046;中国农业大学生物学院农业生物技术国家重点实验室,北京,100094 |
| 基金项目： | 国家高技术研究与发展计划(863)项目(No.2003AA241110和No.2004AA213102)，国家自然科学基金项目(No.30271220)资助 |
| 摘 要： | 选择质粒保有率高及最适合发酵的TOP10菌株,在传统的煮沸裂解法提取质粒的基础上进行了提取方法的改进,使用恒流泵和热交换螺旋管,使菌体在流动过程中完成热裂解,释放质粒DNA。改进后的方法裂解温度可降到70",提取工艺流程简单,缩短了提取时间,提高了质粒DNA的提取效率。利用本方法提取的质粒大小正确、纯度合格,每升发酵液得率平均为100mg质粒DNA。 |
| 关 键 词： | 连续 热裂解 质粒DNA |
| 文章编号： | 1006-1304（2006）01-0051-04 |
| 收稿时间： | 2005-01-10 |
| 修稿时间： | 2005-02-24 |
Large-Scale Plasmid DNA Preparation by Flow-Through Thermal Lysis |
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| ZHU Kai-Chun,ZHANG Fu-Chun,XIAO Chong,HE Zhong-Huai,JIN Hua-Li,WANG Bin. Large-Scale Plasmid DNA Preparation by Flow-Through Thermal Lysis[J]. Journal of Agricultural Biotechnology, 2006, 14(1): 51-54 | |
| Authors: | ZHU Kai-Chun ZHANG Fu-Chun XIAO Chong HE Zhong-Huai JIN Hua-Li WANG Bin |
| Affiliation: | 1.Key Laboratory for Bioresources and Bioengineering, College of Life Science and Technology, Xinjiang University, Urmuqi 830046, China; 2.State Key Laboratory for Agro-biotechnology, College of Biological Science, China Agricultural University, Beijing 100094, China |
| Abstract: | A streamline process of plasmid extraction were developed based on the modified boiling lysis in order to simplify the operation and process for a large volume of Escherichia coli culture. E. coli strain Top10 was selected and used in fermentation because of its stability for hosting plasmids and consistency in the extraction process. The employment of a constant flow pump and a thermal exchange coil enables the lysis step controllable. The size and the purity of plasmids obtained from the process were confirmed and acceptable. Using this method, an average at 100 mg of plasmid could be achieved from one liter fermented culture. |
| Keywords: | flow-through thermal lysis plasmid DNA |
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