Ito-Harashima Sayoko, Yagi Takashi
Department of Biological Science, Graduate School of Science, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai city, Osaka 599-8531, Japan.
J Pestic Sci. 2021 Feb 20;46(1):29-42. doi: 10.1584/jpestics.D20-079.
A reporter gene assay (RGA) is used to investigate the activity of synthetic chemicals mimicking the molting hormones (MHs) and juvenile hormones (JHs) of insects, so-called insect growth regulators (IGRs). The MH receptor, a heterodimer of the ecdysone receptor (EcR) and ultraspiracle (USP), and the JH receptor Methoprene-tolerant (Met) are ligand-dependent transcription factors. Ligand-bound EcR-USP and Met bind to specific -acting DNA elements, referred to as the ecdysone-responsive element (EcRE) and the JH-responsive element (JHRE), respectively, in order to transactivate target genes. Insect hormone-induced transactivation systems have been reconstituted by the introduction of reporter genes under the control of EcRE and JHRE, or two-hybrid reporter genes, into insect, mammalian, and yeast cells expressing receptor proteins. RGA is easy to use and convenient for examining the MH- and JH-like activities of synthetic chemicals and is suitable for the high-throughput screening of novel structural classes of chemicals targeting EcR-USP and Met.
报告基因检测(RGA)用于研究模拟昆虫蜕皮激素(MHs)和保幼激素(JHs)的合成化学物质的活性,即所谓的昆虫生长调节剂(IGRs)。MH受体是蜕皮激素受体(EcR)和超气门蛋白(USP)的异二聚体,JH受体耐甲氧普烯(Met)是配体依赖性转录因子。配体结合的EcR-USP和Met分别与特定的顺式作用DNA元件结合,这些元件分别称为蜕皮激素反应元件(EcRE)和JH反应元件(JHRE),以反式激活靶基因。通过将受EcRE和JHRE控制的报告基因或双杂交报告基因引入表达受体蛋白的昆虫、哺乳动物和酵母细胞中,重建了昆虫激素诱导的反式激活系统。RGA易于使用,便于检测合成化学物质的类MH和类JH活性,适用于针对EcR-USP和Met的新型结构化学物质的高通量筛选。