Help!
I've put GFP onto the C terminus of my protein and expressed it in
planta. It retains biological activity but I can't detect fluoresence.
I've done western blots with antibodies to both GFP and to my protein of
interest. I can't detect anything with the GFP antibodies but detect a
smaller than expected product with the Ab's to my protein so it looks
like the protein is truncated. BUT, he sequence of the clone checks out
and I've successfully used this GFP vector to make other fusion proteins
so it seems to be specific to y p.o.i.. Anyone seen a phenomenon like
this? I'm starting to troubleshoot and thought I would start by
increasing the linker. Any other ideas? Any insights would be much
appreciated. Thanks!
Brendan Riely
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