for my Ph.D. project, I would very much like to create a GFP tagged
receptor protein in c.elegans.
The receptor is a single pass transmembrane protein having it's
C-terminus in the cytoplasm.
I tried to insert my GFP (Andy Fire's) on the N-terminus after the
signal peptide needed for translocation into the ER:
N - sig pep ----- GFP ----- TMD ------- COO-
for unknown reason, this protein ends up in the cytosol and not in the
membrane, as seen by anti-GFP AB on
western blott. I would be very happy to find out if anyone has attempted
to create a similar construct before, and
if it worked :)
hoepfner at mpi-cbg.de