Dear Julie,
I remember there to be a cryptic intron that didn't allow GFP expression in
arabidopsis.
You might check http://ww.google.com/search?q=gfp+cryptic+intron
Regards,
Wo
> Hello
>> We have been using the Clontech EGFP from the vector N2-EGFP to make
> C-terminal fusions with a variety of plasmamembrane located proteins,
> followed by Agrobacterium rhizogenes transformation to produce transgenic
> roots of Medicago truncatula (a plant similar to alfalfa).
> In all cases (nine different constructs) the major GFP protein by Westerns
> is not the fusion but a protein similar in size to EGFP.
> As this EGFP protein is either in the ER or vacuole (rather than the
> cytosol/nucleus - the normal destination of EGFP alone) it seems that the
> fusions are probably produced and target into the ER. However our fusions
> then appear to be degraded to leave mainly the GFP with varying levels of
> the fusion.
>> Have other persons had this problem with GFP fusions?
> Is it particular to i) membrane targeting fusions ii) our plant, iii) the
> GFP and vector that I am using?
>> Many thanks for any help
>> Julie
>>>>> Dr Julie Cullimore
> Laboratoire de Biologie Moléculaire
> des Relations Plantes-Microorganismes,
> CNRS-INRA UMR 215,
> BP 27,
> 31326 Castanet-Tolosan Cedex,
> France
>> Tel: 33 5 61 28 53 22 / 55 13
> Fax: 33 5 61 28 50 61
> Web site: http://www.toulouse.inra.fr/lbmrpm/eng/lbmrpm.htm> E-mail: cullimor at toulouse.inra.fr>> ---
>>
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Dr. Wolfgang Schechinger
Institute of Biomedical Sciences
Academia Sinica, Taipei, Taiwan R.o.C.
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