Stanewsky Ralf
School of Biological and Chemical Sciences, Queen Mary University of London, UK.
Methods Mol Biol. 2007;362:131-42. doi: 10.1007/978-1-59745-257-1_9.
The study of circadian clock function in Drosophila relies heavily on the analysis of rhythmic gene expression. Typically, individuals or groups of flies collected during a specific time of the circadian day need to be sacrificed, followed by the extraction of clock gene products. This procedure makes it impossible to analyze molecular rhythms in an individual over time. To measure clock gene expression within the living animal, firefly luciferase can be used as real-time reporter gene. This chapter describes how rhythmic expression of clock or clock-controlled genes can be measured in living adult Drosophila. A survey of all existing clock-related luciferase transgenics is given.
对果蝇生物钟功能的研究在很大程度上依赖于对节律性基因表达的分析。通常,需要处死在昼夜节律的特定时间收集的果蝇个体或果蝇群体,然后提取生物钟基因产物。这个过程使得无法随时间分析单个个体的分子节律。为了测量活体动物体内的生物钟基因表达,可以使用萤火虫荧光素酶作为实时报告基因。本章描述了如何在成年活体果蝇中测量生物钟或生物钟控制基因的节律性表达。还对所有现有的与生物钟相关的荧光素酶转基因进行了综述。