Taton Arnaud, Ma Amy T, Ota Mizuho, Golden Susan S, Golden James W
Division of Biological Sciences, University of California San Diego , 9500 Gilman Drive, La Jolla, California 92093, United States.
ACS Synth Biol. 2017 Dec 15;6(12):2175-2182. doi: 10.1021/acssynbio.7b00203. Epub 2017 Aug 21.
To downregulate gene expression in cyanobacteria, we constructed NOT gate genetic circuits using orthogonal promoters and their cognate repressors regulated translationally by synthetic riboswitches. Four NOT gates were tested and characterized in five cyanobacterial strains using fluorescent reporter-gene assays. In comparison to alternative systems used to downregulate gene expression in cyanobacteria, these NOT gates performed well, reducing YFP reporter expression by 4 to 50-fold. We further evaluated these NOT gates by controlling the expression of the ftsZ gene, which encodes a prokaryotic tubulin homologue that is required for cell division and is essential for Synechococcus elongatus PCC 7942. These NOT gates would facilitate cyanobacterial genetic engineering or the study of essential cellular processes.
为了下调蓝藻中的基因表达，我们构建了非门遗传电路，该电路使用正交启动子及其同源阻遏物，这些阻遏物由合成核糖开关进行翻译调控。使用荧光报告基因检测法在五种蓝藻菌株中测试并表征了四个非门。与用于下调蓝藻中基因表达的其他系统相比，这些非门表现良好，可将黄色荧光蛋白（YFP）报告基因的表达降低4至50倍。我们通过控制ftsZ基因的表达进一步评估了这些非门，ftsZ基因编码一种原核微管蛋白同源物，细胞分裂需要该同源物，并且对聚球藻PCC 7942至关重要。这些非门将有助于蓝藻基因工程或细胞基本过程的研究。
