Development of a Fluorescent Protein Based FRET Biosensor for Determination of Protease Activity

Proteases are closely associated with many pathological conditions. Efficient detection of protease activity may be useful for diagnosis, prognosis, and the development new therapeutic biomolecules. Fluorescent Resonance Energy Transfer (FRET) is defined as non-radioactive energy transfer that occurs between two fluorophores. proteins widely used in FRET biosensors because they can genetically encoded compatible cells. Protein based (FP-FRET) to monitor biological processes such enzyme activity, intracellular ion concentration, conformational changes, protein-protein interactions. In this study, it was aimed detect using an FP-FRET biosensor TEV chosen a model enzyme. The plasmid encoding mNeonGreen-mRuby3 fluorescent protein-based constructed. gene designed expressed Escherichia coli DH5? cells recombinant DNA techniques purified Ni-NTA affinity chromatography. As result, determined by emission measurements performed spectrofluorometer produced biosensor. usability determination demonstrated.