I am expressing GFP (mut3b version from Cormack et al, Gene 173, 1996) in
trypanosomes. I see excellent expression by fluorescence microscopy and
western blot analysis when I use just GFP. However, with two different
constructs that each add 35 amino acids to the C-terminus of GFP I see no
fluorescence by fluorescence microscopy, even though the fusion proteins
are abundantly expressed based on western blots. One of the C-terminal
"tails" is quite hydrophobic while the other is not.
Has anyone else experienced problems with C-terminal GFP fusions, either
in general, or with short and/or H-phobic peptides?
Any comments or suggestions would be appreciated