Sagot I, Bonneu M, Balguerie A, Aigle M
Laboratoire de Biologie Cellulaire de la Levure, Institut de Biochimie et Génétique Cellulaires, CNRS UPR 9026, Bordeaux, France.
FEBS Lett. 1999 Mar 19;447(1):53-7. doi: 10.1016/s0014-5793(99)00258-6.
We show that fluorescence resonance energy transfer between two mutants of the green fluorescent protein (GFP) can be monitored by imaging microscopy in living yeast. This work is based on the constitutive expression of a GFP-containing fusion protein and the inducible expression of the tobacco etch virus (TEV) protease. In the fusion protein, the P4.3 GFP mutant is linked to the YS65T GFP mutant by a spacer bearing the TEV protease-specific cleavage site.
我们证明，绿色荧光蛋白（GFP）的两个突变体之间的荧光共振能量转移可以通过活酵母中的成像显微镜进行监测。这项工作基于含GFP融合蛋白的组成型表达和烟草蚀纹病毒（TEV）蛋白酶的诱导型表达。在融合蛋白中，P4.3 GFP突变体通过带有TEV蛋白酶特异性切割位点的间隔区与YS65T GFP突变体相连。
