GFP localization ?
a8803349.nospam at unet.univie.ac.at
Fri Nov 27 04:56:33 EST 1998
On 27 Nov 1998 06:09:54 -0000, "Wolfgang Schechinger"
<Wolfgang.Schechinger at med.uni-tuebingen.de> wrote:
>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.
>usual disclaimers apply * This message is RNAse free - please don't touch!
>University of Tuebingen, Germany
>email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
As far as I am informed, there is a size limitation for spontaneous
diffusion into the nucleus of about 70kD (?). GFP is about 35kD,
therefore it is able to spontaneous diffuse into the nucleus. Why it
is brighter there than in the cytoplasm I cant explain.
Please let me know, if you receive any information about this problem!
Internal Med.I,Dept. Oncology
University of Vienna
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