Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae.
Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae.
复制标题
利用强诱导型 DDI2 启动子控制酿酒酵母中的基因表达。
DOI：
10.3389/fmicb.2018.02736
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发表时间：
2018
影响因子：
5.2
通讯作者：
Xiao W
中科院分区：
文献类型：
--
作者：
Lin A;Zeng C;Wang Q;Zhang W;Li M;Hanna M;Xiao W
调控目的基因的表达是酵母研究中常用的方法。在酿酒酵母中，有几种广泛使用的调控表达系统，如GAL和Tet-off系统。然而，所有现有的表达系统都存在一些固有的缺陷。我们以前曾报道过，DDI 2基因可以被诱导到非常高的水平后，氰胺或甲基甲磺酸酯处理。在这里，我们报告的基础上的DDI 2启动子在单拷贝和多拷贝质粒的基因表达系统的建设。使用GFP作为报告基因，证明了通过利用上述系统，靶基因表达可以在转录水平上增加高达2，000倍。此外，创建了基于DDI 2的构建体用于芽殖酵母基因组中的启动子改组以控制内源基因表达。总之，本研究为控制芽殖酵母中目的基因的表达提供了一套方便、高效的实验工具。
Regulating target gene expression is a common method in yeast research. In Saccharomyces cerevisiae, there are several widely used regulated expression systems, such as the GAL and Tet-off systems. However, all current expression systems possess some intrinsic deficiencies. We have previously reported that the DDI2 gene can be induced to very high levels upon cyanamide or methyl methanesulfonate treatment. Here we report the construction of gene expression systems based on the DDI2 promoter in both single- and multi-copy plasmids. Using GFP as a reporter gene, it was demonstrated that the target gene expression could be increased by up to 2,000-fold at the transcriptional level by utilizing the above systems. In addition, a DDI2-based construct was created for promoter shuffling in the budding yeast genome to control endogenous gene expression. Overall, this study offers a set of convenient and highly efficient experimental tools to control target gene expression in budding yeast.