Plieth Christoph
Zentrum für Biochemie und Molekularbiologie, Christian-Albrechts-Universitat, Kiel, Germany.
Methods Mol Biol. 2006;323:307-27. doi: 10.1385/1-59745-003-0:307.
Reporter proteins allow one to monitor cellular parameters that are involved in signal transduction, development, metabolic processes, and transport. There are targeting strategies available to direct the indicator protein exactly to the locale inside the organism from which information is desired. This circumvents experimental reductionism and allows experimentation with whole intact and undisturbed organisms. The outstanding advantages of self-reporting organisms make it worth to shoulder cost- and time-consuming molecular work. Here, the luminescent Ca2+ indicator aequorin is introduced and a rough guideline is given from early planning the molecular work and assembling an experimental setup to experimentation with luminescent Arabidopsis, data processing, and control experiments.
报告蛋白使人们能够监测参与信号转导、发育、代谢过程和运输的细胞参数。有一些靶向策略可用于将指示蛋白精确地引导到生物体内需要获取信息的部位。这避免了实验还原论,并允许对完整无损的生物体进行实验。自我报告生物体的显著优势使得进行耗时且成本高昂的分子工作是值得的。在此,引入了发光钙指示剂水母发光蛋白,并给出了一个大致的指导方针,内容涵盖从早期分子工作规划、组装实验装置到对发光拟南芥进行实验、数据处理以及对照实验等方面。