| 论文题目: | Bacterial Genome Editing via a Designed Toxin-Antitoxin Cassette |
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| 作者: | Wu Jie, Deng Aihua, Sun Qinyun, Bai Hua, Sun Zhaopeng, Shang Xiuling, Zhang Yun, Liu Qian, Liang Yong, Liu Shuwen, Che Yongsheng, and Wen Tingyi* |
| 联系作者: | |
| 刊物名称: | ACS Synth Biol |
| 期: | 7 |
| 卷: | 3 |
| 页: | 822-831 |
| 年份: | 2018 |
| 影响因子: | 5.253 |
| 论文下载: | 下载地址 |
| 摘要: | Manipulating the bacterial genomes in an efficient manner is essential to biological and biotechnological research. Here, we reprogrammed the bacterial TA systems as the toxin counter-selectable cassette regulated by an antitoxin switch (TCCRAS) for genetic modifications in the extensively studied and utilized Gram-positive bacteria, B. subtilis and Corynebacterium glutamicum. In the five characterized type II TA systems, the RelBE complex can specifically and efficiently regulate cell growth and death by the conditionally controlled antitoxin RelB switch, thereby serving as a novel counter-selectable cassette to establish the TCCRAS system. Using a single vector, such a system has been employed to perform in-frame deletion, functional knock-in, gene replacement, precise point mutation, large-scale insertion, and especially, deletion of the fragments up to 194.9 kb in B. subtilis. In addition, the biosynthesis of lycopene was first achieved in B. subtilis using TCCRAS to integrate a 5.4-kb fusion cluster ( P spac- crtI- crtE- crtB). The system was further adapted for gene knockdown and replacement, and large-scale deletion of the fragments up to 179.8 kb in C. glutamicum, with the mutation efficiencies increased by 0.8-1.0-fold compared to the conventional SacB method. TCCRAS thus holds promise as an effective and versatile genome-scale engineering technology for metabolic engineering and synthetic genomics research in a broad range of the Gram-positive bacteria. |
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