State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100101, China.
Sci Rep. 2019 Aug 29;9(1):12538. doi: 10.1038/s41598-019-49000-8.
Tuned gene expression is crucial to the proper growth and response to the environmental changes of an organism. To enable tunable gene expression as designed is desirable in both scientific research and industrial application. Here, we introduce a novel promoter switching method based on the DDI2 promoter (P) that can fine tune the expression of target genes. We constructed a recyclable cassette (P-URA3-P) and integrated it upstream of yeast target genes to replace the native promoters by DDI2 promoter without introducing any junk sequence. We found that the presence or absence of cyanamide as an inducer could turn on or off the expression of target genes. In addition, we showed that P could act as a gene switch to linearly regulate the expression levels of target genes in vivo. We switched the original promoters of RAD18, TUP1, and CDC6 with P as a proof-of-concept.
基因表达的调控对于生物的正常生长和对环境变化的响应至关重要。在科学研究和工业应用中，都期望能够实现可调控的基因表达。在这里，我们介绍了一种基于 DDI2 启动子（P）的新型启动子切换方法，该方法可以精细调节靶基因的表达。我们构建了一个可回收的盒式结构（P-URA3-P），并将其整合到酵母靶基因的上游，通过 DDI2 启动子取代天然启动子，而不引入任何无用序列。我们发现，氰氨作为诱导剂的存在与否可以开启或关闭靶基因的表达。此外，我们还表明，P 可以作为一种基因开关，在线性调节靶基因在体内的表达水平。我们将 RAD18、TUP1 和 CDC6 的原始启动子替换为 P，作为概念验证。
