| 大肠杆菌基因无痕敲除技术及策略 |
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| 引用本文： | 胡逢雪,丁锐,崔震海,于金龙,刘丽霏,敖永华,张立军. 大肠杆菌基因无痕敲除技术及策略[J]. 生物技术通讯, 2013, 0(4): 552-557 |
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| 作者姓名： | 胡逢雪 丁锐 崔震海 于金龙 刘丽霏 敖永华 张立军 |
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| 作者单位： | [1]沈阳农业大学生物科学技术学院,辽宁沈阳110866 [2]沈阳农业大学工程学院,辽宁沈阳110866 [3]辽宁省植物基因工程技术研究中心,辽宁沈阳110866 [4]哈佛医学院实验医学部,美国波士顿02215 |
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| 基金项目： | 国家自然科学基金（30870190）;教育部博士点基金（200801570005）;沈阳市科技攻关项目（1091104-3-04）;辽宁省科技型中小企业技术创新专项（201205635） |
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| 摘 要： | 基因敲除技术是大肠杆菌基因组减小和代谢途径改造的有效手段，其中基于同源重组原理的基因无痕敲除技术显现出其他技术所不具备的应用优势和发展潜力。该技术可以快速敲除大肠杆菌基因组中的目标基因，并且在基因组中不残留任何外源片段，所以不会干扰后续的基因操作。我们分类介绍了无痕敲除技术中所涉及载体的结构、功能及其相应的敲除策略，着重介绍了无痕敲除技术的原理及载体构建方法。
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| 关 键 词： | 大肠杆菌 基因无痕敲除 基因替换 Red重组系统 代谢工程 |
Approaches and Strategies of Gene Scarless Knockout in the Esche-richia coli Genome |
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| HU Feng-Xue,DING Rui,CUI Zhen-Hai,YU Jin-Long,LIU Li-Fei,AO Yong-Hua,ZHANG Li-Jun. Approaches and Strategies of Gene Scarless Knockout in the Esche-richia coli Genome[J]. Letters in Biotechnology, 2013, 0(4): 552-557 |
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| Authors: | HU Feng-Xue DING Rui CUI Zhen-Hai YU Jin-Long LIU Li-Fei AO Yong-Hua ZHANG Li-Jun |
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| Affiliation: | 1. a. College of Biological Sciences and Technology; b. College of Engineering; Shenyang Agricultural University, Shenyang 110866, China; 2. Liaoning Province Research Center of Plant Gene Engineering Technology, Shenyang 110866, China; 3. Division of Experimental Medicine, Beth Israel Deaconess Medical Center, Halward Medical School, Boston, Massachusetts 02215, USA) |
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| Abstract: | Gene knockout has been broadly applied in genome reduction and metabolic engineering of Escherich-ia coli. The scarless deletion technology based on homologous recombination provides clear application advantages over other knockout methods. This technology allows rapid and sequential deletion of targeted genes without intro-duction of any exogenous sequences in the genome. In this article we presented the structures and functions of dif-ferent types of vectors applied in scarless deletion, the principle and the procedure of scarless knockout technolo-gy were also described in detail. |
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| Keywords: | Escherichia coli scarless gene deletion gene replacement λ Red recombination system metabolicengineering |
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