We designed and tested a modular epitope system for genomic tagging of the proteins. Our modular tag contains one of the versions of a fluorescent protein for in vivo localization and a small double affinity tag for immunopurification.
A modular approach allows us to decoupled the two types of experiments. We do not rely on the availability of a good antibody against a fluorescent protein for affinity purification. Instead we focus on optimizing the performance of each module for studying the dynamics of protein localization and interactions.
Here is a set of vectors we developed for tagging the proteins with a modular tag. The tag contains one of the versions of a monomeric fluorescent protein (GFP (S65T), EYFP, mOrange, mCitrine, Cherry etc.) fused at the C-terminus to a small double affinity tag for immunopurification (3xFLAG-6xH, 3xMyc-8xH, 4Strep-8xH, etc). The sequences of the vectors can be found here.
