GFP localization ?

[Paul Kitts]

Wolfgang,
Unfused GFP is small enough to diffuse in and        |
out through the nuclear pores.                       V 
Unfused GFP can appear to be localized to the        _
nucleus simply because you are looking at the     __/0\__
fluorescence from a greater depth of cell.       ---------
                                                                   
Some people have reported that the apparent distribution of unfused GFP varies 
with cell type (see previous postings to this group 
http://www.bio.net/hypermail/FLUORESCENT-PROTEINS). This may reflect differences 
in the cell morpology.
-=Paul=-

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"  Paul Kitts, Ph.D.               NCBI/NLM/NIH                "
"  Email: kitts at ncbi.nlm.nih.gov   Bldg 38A, 8N803             "
"  Phone: 301 435-6091             8600 Rockville Pike         "
"  Fax:   301 480-9241             Bethesda, MD 20894, U.S.A.  "
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--------------------------------------------------------------------
To: fluorpro at net.bio.net
From: Wolfgang.Schechinger at med.uni-tuebingen.de ("Wolfgang Schechinger")
Subject: GFP localization ?
Date: 26 Nov 1998 22:13:48 -0800

Hi all, 

When transfecting 293HEK and NIH 3T3 MF-cells with GFP (humanized 
S65T in pCDNA3) I noticed the brightest fluorescence in the nucleus, 
while a (certain) fusion protein doesn't. Is there a reason for this? 
Actually, I had expected that there was no fluorescence in the 
nucleus since there's no nuclear import sequence. Am I wrong? Is 
there resonance energy transfer from DNA to GFP? 
The phenomenon occurs when using a Hg lamp with filters for FITC as 
well as a 488nm argon laser and a confocal microscope.

Any ideas?

Wolfgang
-----
usual disclaimers apply * This message is RNAse free - please don't touch!
----- 
Wolfgang Schechinger 
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: 
http://www.medizin.uni-tuebingen.de/~wgschech/start.htm