Fluoromodules are composed of biomolecules that generate strong fluorescence upon interacting with their associated fluorogenic dye. Previous work on genetically encodable fluoromodules has developed fluorescence activating protein (FAP)- dye pairs with immediate application in tracking proteins at the cell surface. A new thiazole orange derivative substituted with an electron-withdrawing group exhibits low DNA binding and high photostability, making it a promising candidate for selection of a protein-binding partner in order to create a new green fluoromodule. We have used our streamlined fluorescence activated cell sorting (FACS) method for identifying candidate proteins from a yeast surface-displayed library. Six rounds of FACS enrichment led to isolation of several unique clones that exhibit high dye binding affinity and bright green fluorescence.

Genetically encodable green fluorescent dye-protein complexes with improved photostability

243rd National Meeting (2012)

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