| In this paper, the study of the interaction of protein-protein about cellular proliferation and apoptosis signal pathway by the fluorescence resonance energy transfer (FRET) technology have been given.The first, this paper introduces synoptically that the application of FRET technology in the study of protein-protein interaction and the principle of the laser confocal scan microscope. The paper summurized the evolution of the application of FRET technology in the study of protein-protein interaction.The cells lysis is necessary for detection of PKA activity with traditional means, then the PKA activity is detected in the test tube, which has the limitation. We present a genetically encoded fluorescent reporter for protein kinase A (PKA) consisting of fusions of cyan fluorescent protein, a phosphoamino acid binding domain (14-3-3), a consensus substrate for PKA, and yellow fluorescent protein. cAMP elevations cause 25-50% changes in the ratios of yellow to cyan emissions in live cells caused by phosphorylationinduced changes in fluorescence resonance energy transfer. This technology offsets the limitation of tranditional means.In the recent work, we constructed a vector that 14-3-3 fused with yellow fluorescent protein(YFP). The other vector-pBid-CFP was provided by Professor Kazunari Taira of University of Tokyo. Binding of tBid and 14-3-3 alters the distance between CFP and YFP and generates a FRET change, so the binding of 14-3-3 and tBid will be rightly reflected by FRET change between CFP and YFP. Using our FRET system, we obtained more true information of the interaction of 14-3-3 and tBid as well as in vivo images of Bid-CFP and YFP-14-3-3 respectively that make us clearly in spatio-temporal dynamic process of the interaction of 14-3-3 and tBid during TNF-α-induced cellular apoptosis.At last, the foreground of this study is prospected. We think that FRETtechnology can reflect the interaction of protein-protein in the living cells, and it has very good evolution prospect.
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