I'm trying to maximise GFP-signal in my yeast cells to detect relatively low
levels of protein expressed from a weak promoter and consequently have a few
quick questions. I've read on this newsgroup that people have successfully
fused 2 in-frame copies of GFP to produce an even brighter reporter
molecule. Could anyone tell me what the optimal length of linker between
two such GFP monomers is ? and what (if any) are the amino-acid sequence
requirements ? Or where I can even find this information ?
A major problem is likely to be signal-masking by the cell wall and
therefore another experiment I'm considering is to add a secretory signal to
GFP - does anybody know if there is a size limit on proteins to be secreted
from yeast ?
Thanks in advance !
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