Periasamy A, Day R N
J Biomed Opt. 1998 Apr;3(2):154-60. doi: 10.1117/1.429872.
The combined use of fluorescence resonance energy transfer (FRET) microscopy and expression of genetic vectors encoding protein fusions with green fluorescent protein (GFP) and blue fluorescent protein (BFP) provides an exceptionally sensitive method for detecting the interaction of protein partners in living cells. The acquisition of FRET signals from GFP- and BFP-fusion proteins expressed in living cells was demonstrated using an optimized imaging system and high sensitivity charge coupled device camera. This imaging system was used to detect energy transfer signals from a fusion protein containing GFP physically linked to BFP expressed in living HeLa cells. In contrast, the co-localization of noninteracting GFP- and BFP-fusion proteins was not sufficient for energy transfer. The FRET imaging system was then used to demonstrate dimerization of the pituitary-specific transcription factor Pit-1 within the living cell nucleus. © 1998 Society of Photo-Optical Instrumentation Engineers.
荧光共振能量转移（FRET）显微镜与编码绿色荧光蛋白（GFP）和蓝色荧光蛋白（BFP）的蛋白质融合体的基因载体表达相结合，为检测活细胞中蛋白质伴侣间的相互作用提供了一种极其灵敏的方法。利用优化的成像系统和高灵敏度电荷耦合器件相机，证明了从活细胞中表达的GFP和BFP融合蛋白获取FRET信号的可行性。该成像系统用于检测来自在活的HeLa细胞中表达的、物理连接有BFP的GFP融合蛋白的能量转移信号。相比之下，不相互作用的GFP和BFP融合蛋白的共定位不足以实现能量转移。然后，利用FRET成像系统证明了垂体特异性转录因子Pit-1在活细胞核内的二聚化。© 1998 美国光电仪器工程师协会。
