Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology.
School of Life Science and Technology, Tokyo Institute of Technology.
J Gen Appl Microbiol. 2021 Apr 16;67(1):42-46. doi: 10.2323/jgam.2020.02.003. Epub 2020 Sep 16.
The firefly luciferase (Luc) reporter assay is a powerful tool used to analyze promoter activities in living cells. In this report, we established a firefly Luc reporter assay system in the unicellular model red alga Cyanidioschyzon merolae. A nitrite reductase (NIR) promoter-Luc fusion gene was integrated into the URA5.3 genomic region to construct the C. merolae NIR-Luc strain. Luc activities in the NIR-Luc strain were increased, correlating with the accumulation of endogenous NIR transcripts in response to nitrogen depletion. Luc activity was also significantly increased by the overexpression of the MYB1 gene, which encodes a transcription factor responsible for NIR promoter activation. Thus, our results demonstrate the utility of the Luc reporter system in C. merolae.
萤火虫荧光素酶（Luc）报告基因检测是一种强大的工具，用于分析活细胞中的启动子活性。在本报告中，我们在单细胞模型红藻衣藻中建立了萤火虫 Luc 报告基因检测系统。将亚硝酸盐还原酶（NIR）启动子-Luc 融合基因整合到URA5.3 基因组区域，构建了衣藻 NIR-Luc 株。NIR-Luc 株的 Luc 活性随着内源 NIR 转录本的积累而增加，与氮饥饿反应相关。MYB1 基因的过表达也显著增加了 Luc 活性，MYB1 基因编码一个负责 NIR 启动子激活的转录因子。因此，我们的结果表明 Luc 报告基因检测系统在衣藻中的实用性。
