Development of caspase-3 indicator based-on bioluminescence-fluorescence double resonant energy transfer

在本研究中，首先，基于生物发光共振能量转移（BRET）的Ca^<2+>指示剂BRAC被开发和测试以评估其理化性质。第二，BRAC被用于观察活细胞（HeLa细胞）和活植物（拟南芥）叶片中的Ca^2+动态。最后，为了制备基于生物发光-荧光双共振能量转移的指示剂，研究了合适的FRET受体为FRET供体Venus的可能的橙子至红色荧光蛋白候选物。然而，金星和这些橙子到红色荧光蛋白之间的FRET效率很差，因此需要进一步的研究。

In this study, first, bioluminescence resonance energy transfer (BRET) based Ca^<2+> indicator, BRAC was developed and tested to evaluate its physiochemical properties. Second, BRAC was used to visualize Ca^<2+> dynamics in living cells (HeLa cells) and living plant (Arabidopsis thaliana) leaf. Finally, to make the indicator based on bioluminescent-fluorescent double resonant energy transfer, appropriate FRET acceptor for FRET donor Venus were investigated in possible orange to red fluorescent protein candidates. However FRET efficiency between Venus and these orange to red fluorescent proteins were poor, so additional studies should be needed.