Dual fluorescent protein-based bioassay system for the detection of genotoxic chemical substances in Saccharomyces cerevisiae

作者：Lu, Yixin; Tian, Yongjie; Wang, Ruikun; Wu, Qianqian; Zhang, Yu; Li, Xiangming^*

来源：Toxicology Mechanisms and Methods, 2015, 25(9): 698-707.

DOI：10.3109/15376516.2015.1070305

• Chemical substances
• DNA damage
• DsRed-Express2 reporter
• genotoxicity
• yeast
• yEGFP reporter

A new reporter system has been developed for quantifying the activity of potentially DNA-damaging substances in the yeast Saccharomyces cerevisiae. The system relies on two different reporter genes, yEGFP and DsRed-Express2, to screen for DNA-damaging chemicals. The yEGFP gene is fused to the test promoter of RNR2, whose measurable signal has a dose-dependent relationship with DNA damage. The gene encoding DsRed-Express2 is fused to a constitutive promoter of GPD, providing an internal control for normalizing cell numbers in the assay. The dual fluorescent protein assay system is performed by sequentially measuring the yEGFP and DsRed-Express2 fluorescent intensity of the same sample, with the results expressed as the ratio of yEGFP to DsRed-Express2 intensity (yEGFP/DsRed-Express2). The yeast fluorescent protein reporter assay was performed in 96-well microtiter plates in the presence of different concentrations of test substances, which were then characterized. The assay was very efficient, high-throughput, and amenable to full automation. Here, we demonstrate that this system can be used as a biosensor to assess the genotoxic potential of drugs and other chemical substances.

• 出版日期2015-11-22
• 单位扬州大学