Is there a difference between tagging GFP at the N-terminus and at the C-terminus of a gene?
Posted April 24, 2020
Tagging at the N-terminus instead of the C-terminus greatly increases the risk of the protein not localizing correctly, due to masking of the signal sequence at the N-terminus. Tagging at the C-terminus is thus better for preserving localization; an exception to this, however, is for a handful proteins that localize to cell walls or the ER. These often contain C-terminal signal sequences, which can be interrupted by C-terminal tagging.
Palmer, E., & Freeman, T. (2004). Investigation into the use of C- and N-terminal GFP fusion proteins for subcellular localization studies using reverse transfection microarrays. Comparative and functional genomics, 5(4), 342–353. doi:10.1002/cfg.405