Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae

Abstract

Regulating target gene expression is a common method in yeast research. In <i>Saccharomyces cerevisiae</i>, there are several widely used regulated expression systems, such as the <i>GAL</i> and Tet-off systems. However, all current expression systems possess some intrinsic deficiencies. We have previously reported that the <i>DDI2</i> 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 <i>DDI2</i> promoter in both single- and multi-copy plasmids. Using <i>GFP</i> 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 <i>DDI2</i>-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.