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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