Oakenfull Rachael J, Ronald James, Davis Seth J
Department of Biology, University of York, York, UK.
Methods Mol Biol. 2019;2026:179-192. doi: 10.1007/978-1-4939-9612-4_15.
The circadian clock allows plants to synchronize their internal processes with the external environment. This synchronization occurs through daily cues, one of which is light. Phytochromes are well established as light-sensing proteins and have been identified in forming multiple signaling networks with the central circadian oscillator. However, the precise details of how these networks are formed are yet to be established. Using established promoter-luciferase lines for clock genes crossed into mutant lines, it is possible to use luciferase-based imaging technologies to determine whether specific proteins are involved in phytochrome signaling to the circadian oscillator. The methods presented here use two automated methods of luciferase imaging in Arabidopsis to allow for high-throughput measurement of circadian clock components under a range of different light conditions.
生物钟使植物能够将其内部过程与外部环境同步。这种同步通过每日的信号线索来实现，其中之一就是光。光敏色素作为光感应蛋白已被充分确立，并且已被确定在与中央生物钟振荡器形成多个信号网络中发挥作用。然而，这些网络如何形成的精确细节尚未确定。利用已建立的用于时钟基因的启动子-荧光素酶系与突变体系杂交，可以使用基于荧光素酶的成像技术来确定特定蛋白质是否参与了向生物钟振荡器的光敏色素信号传导。这里介绍的方法使用了拟南芥中两种荧光素酶成像的自动化方法，以便在一系列不同光照条件下对生物钟组件进行高通量测量。
