Suzuki Nobuaki, Okayama Satoshi, Nonaka Hiroshi, Tsuge Yota, Inui Masayuki, Yukawa Hideaki
Microbiology Research Group, Research Institute of Innovative Technology for Earth (RITE), 9-2, Kizugawadai, Kizu-Cho, Soraku-Gun, Kyoto 619-0292, Japan.
Appl Environ Microbiol. 2005 Jun;71(6):3369-72. doi: 10.1128/AEM.71.6.3369-3372.2005.
The engineering of Corynebacterium glutamicum is important for enhanced production of biochemicals. To construct an improved C. glutamicum genome, we developed a precise genome excision method based on the Cre/loxP recombination system and successfully deleted 11 distinct genomic regions identified by comparative analysis of C. glutamicum genomes. Despite the loss of several predicted open reading frames, the mutant cells exhibited normal growth under standard laboratory conditions. With a total of 250 kb (7.5% of the genome), the 11 genomic regions were loaded with cryptic prophages, transposons, and genes of unknown function which were dispensable for cell growth, indicating recent horizontal acquisitions to the genome. This provides an interesting background for functional genomic studies and can be used in the improvement of cell traits.
谷氨酸棒杆菌的工程改造对于提高生化物质的产量至关重要。为构建改良的谷氨酸棒杆菌基因组,我们基于Cre/loxP重组系统开发了一种精确的基因组切除方法,并成功删除了通过谷氨酸棒杆菌基因组比较分析鉴定出的11个不同的基因组区域。尽管缺失了几个预测的开放阅读框,但突变细胞在标准实验室条件下仍表现出正常生长。这11个基因组区域总共250 kb(占基因组的7.5%),富含隐秘原噬菌体、转座子和功能未知的基因,这些对于细胞生长是可有可无的,表明它们是最近水平转移到基因组中的。这为功能基因组学研究提供了一个有趣的背景,并且可用于改善细胞特性。