I'm trying to conjugate GFP on my target membrane protein in overexpression plasmid.
While searching for published plasmid data, i came across a plasmid which have inserted GFP after predicted signal peptide cleavage site as anything upstream of that will be cleaved off in ER.
so my question is...
1) is it really necessary to insert my GFP at that specific site of "after predicted signal peptide cleavage site" or isn't it entirely possible to simply insert the GFP at the 3' end of the protein?
2) how would yo go about finding this exact signal peptide cleavage site?
any comments appreciated!