| 一种由寡核苷酸介导的大肠杆菌ftsZ基因无痕敲除的方法 |
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| 引用本文： | 张飞飞,杨运文,董慧青,尚广东. 一种由寡核苷酸介导的大肠杆菌ftsZ基因无痕敲除的方法[J]. 生物技术进展, 2018, 8(2): 174-179. DOI: 10.19586/j.2095-2341.2017.0130 |
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| 作者姓名： | 张飞飞 杨运文 董慧青 尚广东 |
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| 作者单位： | 1.徐州医科大学麻醉学院, 江苏省麻醉学重点实验室, 江苏 徐州 221004;2.南京师范大学生命科学学院, 江苏省微生物与功能基因组学重点实验室, 南京 210023 |
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| 基金项目： | 国家自然科学基金项目(NSFC81273412)；徐州医科大学校级课题（2017KJ08）资助。 |
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| 摘 要： | 现阶段，适用于大肠杆菌的无痕敲除方法普遍存在周期较长、操作步骤复杂等问题。为了进一步改进和优化无痕敲除技术，采用单链寡核苷酸介导的Red同源重组系统（single strand oligonucleotide-mediated recombineering，SSOR），通过两步连续的同源重组，敲除了一种编码类似微管蛋白的GTP酶的ftsZ基因。该方法可快速高效无痕的敲除目的基因，为大肠杆菌基因组改造提供了有效方法，另外，ftsZ基因缺失株的获得也为研究ftsZ基因功能创造了条件。
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| 关 键 词： | ftsZ基因 重组工程 单链寡核苷酸 大肠杆菌 |
| 收稿时间： | 2017-09-22 |
A Scarless Knockout Method of ftsZ Gene in Escherichia coli Mediated by Oligonucleotide |
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| ZHANG Feifei,YANG Yunwen,DONG Huiqing,SHANG Guangdong. A Scarless Knockout Method of ftsZ Gene in Escherichia coli Mediated by Oligonucleotide[J]. CURRENT BIOTECHNOLOGY, 2018, 8(2): 174-179. DOI: 10.19586/j.2095-2341.2017.0130 |
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| Authors: | ZHANG Feifei YANG Yunwen DONG Huiqing SHANG Guangdong |
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| Affiliation: | 1.Jiangsu Province Key Laboratory of Anesthesiology, College of Anesthesiology, Xuzhou Medical University, Jiangsu Xuzhou 221004, China; 2.Jiangsu Key Laboratory for Microbes and Functional Genomics, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China |
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| Abstract: | At present, the scarless knockout technology for Escherichia coli have many shortcomings, such as long period, complex operation steps, etc. In order to further improve and optimize the scarless knockout technology, E. coli tubulin-like GTPase gene ftsZ was knocked out via single strand oligonucleotide-mediated recombineering(SSOR) including two-step successive homologous recombination. The rapid and efficient scarless gene knockout method has provided an effective method for the transformation of E. coli genome. In addition, the gene deletion strain of ftsZ has laid the foundation for the study of ftsZ gene function. |
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| Keywords: | ftsZ gene recombineering single strand oligonucleotide(SSO) Escherichia coli |
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