Hoshino Hideto, Nakajima Yoshihiro, Ohmiya Yoshihiro
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Osaka 563-8577, Japan.
Nat Methods. 2007 Aug;4(8):637-9. doi: 10.1038/nmeth1069. Epub 2007 Jul 8.
Taking advantage of the phenomenon of bioluminescence resonance energy transfer (BRET), we developed a bioluminescent probe composed of EYFP and Renilla reniformis luciferase (RLuc)--BRET-based autoilluminated fluorescent protein on EYFP (BAF-Y)--for near-real-time single-cell imaging. We show that BAF-Y exhibits enhanced RLuc luminescence intensity and appropriate subcellular distribution when it was fused to targeting-signal peptides or histone H2AX, thus allowing high spatial and temporal resolution microscopy of living cells.
利用生物发光共振能量转移(BRET)现象,我们开发了一种由增强型黄色荧光蛋白(EYFP)和海肾荧光素酶(RLuc)组成的生物发光探针——基于BRET的EYFP上的自发光荧光蛋白(BAF-Y),用于近实时单细胞成像。我们发现,当BAF-Y与靶向信号肽或组蛋白H2AX融合时,其RLuc发光强度增强且亚细胞分布合适,从而能够对活细胞进行高时空分辨率的显微镜观察。