Subramanian Chitra, Xu Yao, Johnson Carl Hirschie, von Arnim Albrecht G
Department of Botany, The University of Tennessee, Knoxville, USA.
Methods Mol Biol. 2004;284:271-86. doi: 10.1385/1-59259-816-1:271.
The emerging technique of bioluminescence resonance energy transfer (BRET) allows us to detect protein interactions in live cells and in real time, thus providing a new window into cellular signal transduction processes. We present experimental protocols for expressing fusion proteins between luciferase and fluorescent proteins that are the basis for BRET measurement, as well as for measuring and imaging BRET in a variety of cell types. Despite our focus on plant cells, the techniques described here are easily adaptable to other cell systems that have yet to benefit from the BRET technique.
新兴的生物发光共振能量转移(BRET)技术使我们能够在活细胞中实时检测蛋白质相互作用,从而为细胞信号转导过程提供了一个新的窗口。我们展示了用于表达荧光素酶和荧光蛋白之间融合蛋白的实验方案,这些融合蛋白是BRET测量的基础,同时也展示了在多种细胞类型中测量和成像BRET的方案。尽管我们重点关注植物细胞,但这里描述的技术很容易适用于其他尚未从BRET技术中受益的细胞系统。