Production of intracellular compartiments specific markers for the study of GPCR and g protein trafficking by BRET technology

Abstract

After bibliographic research, we will identify resident proteins from different intracellular compartments, which could be used as markers for such compartments such as Golgi, endoplasmic reticulum, mitochondria and nucleus. After selecting the possible targets, we will construct markers for different intracellular compartments through Molecular Biology techniques, fusing such proteins, or protein localization domains, to different GFP or RLucII isoforms. The constructs will be analyzed by transfecting mammalian cells and evaluating them by fluorescence confocal microscopy for verification and confirmation of correct localization. Finally, we will perform BRET experiments between the new constructs and GPCRs or G proteins of interest, aiming at a better understanding of the Spatio-temporal regulation of such molecules.