Fine-tuning the expression of target genes using a DDI2 promoter gene switch in budding yeast

Fine-tuning the expression of target genes using a DDI2 promoter gene switch in budding yeast
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使用芽殖酵母中的 DDI2 启动子基因开关微调目标基因的表达

DOI:
10.1038/s41598-019-49000-8
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发表时间:
2019-08
期刊:
影响因子:
4.6
通讯作者:
Fu Yu V
Fu Yu V
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Wang Yong;Zhang Kaining;Li Hanfei;Xu Xin;Xue Huijun;Wang Pingping;Fu Yu V

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调节基因表达对于生物体的正常生长和对环境变化的反应是至关重要的。无论是在科学研究还是在工业应用中,都希望能够按照设计实现可调的基因表达。在这里,我们介绍了一种新的基于DDI2启动子的启动子切换方法(PDDI2),它可以微调目的基因的表达。我们构建了一个可回收的盒(PDDI2-URA3-PDDI2),并将其整合到酵母目的基因的上游,以DDI2启动子取代天然启动子,而不引入任何垃圾序列。我们发现,氰胺作为诱导剂的存在或不存在都可以开启或关闭目标基因的表达。此外,我们还发现PDDI2在体内可以作为一个基因开关来线性调节靶基因的表达水平。我们用PDDI2替换了RAD18、Tup1和CDC6的原始启动子作为概念验证。
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 (PDDI2) that can fine tune the expression of target genes. We constructed a recyclable cassette (PDDI2-URA3-PDDI2) 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 PDDI2 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 PDDI2 as a proof-of-concept.
利用强诱导型 DDI2 启动子控制酿酒酵母中的基因表达。
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