TRICISTRONIC VECTOR FOR CONDUCTING PROTEIN INTERACTION ASSAYS THROUGH THE BIMOLECULAR FLUORESCENCE COMPLEMENTATION TECHNIQUE IN MAMMALIAN CELL CULTURE.

摘要

The present disclosure is related to a tricistronic vector to conduct interaction assays through the bimolecular fluorescence complementation technique in mammalian cell culture, the pTRI-BIFC-mCherry vector makes possible the evaluation of BIFC tests more easily and unambiguously, since proteins are expressed from a single vector, resulting in a more efficient cell count. The mCherry fluorescence indicates that the vector entered the cell and the presence or absence of fluorescence of the Venus protein indicates whether or not there is interaction, unlike the conventional triple transfection protocol, wherein fluorescence of said reporter protein is observed, but there is no assurance that the remaining vectors have entered the cell, thereby producing false negatives.