Benedikt,
CFP has a much lower quantum efficiency than GFP or YFP. If your pollen tubes
are growing in the light, maybe the high level of CFP is producing enough free
radicals to affect growth. If this was the problem, the CFP should not affect
the pollen tubes in the dark.
-=Paul=-
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" Paul Kitts, Ph.D. "
" National Center for Biotechnology Information "
" NIH/NLM "
" Bldg. 38A, Rm. 8N803 Email: kitts at ncbi.nlm.nih.gov "
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------------------------------------------------------------------------
To: fluorpro at net.bio.net
From: benedikt at ima.org.sg ("Benedikt Kost")
Subject: CFP toxicity
Date: 4 Oct 1999 21:56:36 -0700
Dear all
I have found that transient expression of CFP (ECFP, clontech) at high
levels affects the growth of tobacco pollen tubes, wheres GFP and YFP
expression does not!
Did anyone observe specific toxicity of CFP (but not GFP and YFP) expression
in any other cell type? If so, did anyone find a way around this? Not all
the point mutations in ECFP are apparently required for the change in
fluorescence characteristics, so maybe reverting some of the mutations to
wild type may reduce toxicity without altering fluorescence excitation and
emission spectra?
I am greatful for any comments!
Thanks,
Benedikt
----------------------------------------------------
Dr. Benedikt Kost
Lab of Plant Cell Biology
Institute of Molecular Agrobiology
National University of Singapore
1, Research Link
Singapore, 117604
tel +65 872 7483
fax +65 872 7007
email benedikt at ima.org.sg
