摘要
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.
摘要译文
谷氨酸棒状杆菌（Corynebacterium glutamicum）的工程化对于提高生物化学物质的产量是重要的。为了构建改良的谷氨酸棒状杆菌基因组，我们开发了基于Cre / loxP重组系统的精确的基因组切除方法，并成功地删除了通过谷氨酸棒状杆菌基因组的比较分析鉴定的11个不同的基因组区域。尽管几个预测的开放阅读框丢失，但突变细胞在标准的实验室条件下显示出正常的生长。总共250kb（基因组的7.5％），11个基因组区域装载有隐藏的预变体，转座子和功能未知的基因，这些基因组对于细胞生长是不必要的，表明最近对基因组的水平获取。这为功能基因组研究提供了一个有趣的背景，可用于改善细胞性状。
Nobuaki Suzuki1; Satoshi Okayama1;2; Hiroshi Nonaka1; Yota Tsuge1;2; Masayuki Inui1 ; Hideaki Yukawa1;2;*. Large-Scale Engineering of the Corynebacterium glutamicum Genome[J]. Applied and Environmental Microbiology, 2005,71(6): 3369-3372