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Replies to accumulation of GFP in plant nucleus

Alvar Carlson ACARLSON at CROP.UOGUELPH.CA
Tue Apr 29 13:58:55 EST 1997


On April 14th I posted the following questions:
>
>    I am attempting to use sGFP to visually select transgenic barley. 
>I am concerned that the accumulation of the GFP in the nucleus will 
>be detrimental to the development of transgenic plants.  Has anyone 
>observed reduced transformation efficiency in plants when using GFP 
>or reduced fertility in any of the transgenic plants generated?  
>Furthermore, has anyone found that modified GFP, to exclude it from 
>the nucleus, increases the transformation efficiency?
.........................................


From:             David Galbraith <dgalbrai at ag.Arizona.EDU>

We have expressed GFP transgenically targeted to the nucleus in tobacco and
have seen no evidence of toxicity.  Plants have not gone to the next
generation yet.

From:             sjdavis1 at students.wisc.edu (Seth J. Davis)

WT GFP is toxic, and interferes with transformation efficiency.  Modified
GFPs (smGFP or mGFP5) appear to be less toxic in regenerating plants.
mGFP5, which is ER localized (not in the nucleus) might reduce the toxic
effects of GFP.  This last point is not clear; is making GFP soluble or is
localizing GFP to the ER increasing transformation efficiency?  This has
not been tested.

From:             krs1 at mrc-lmb.cam.ac.uk (Kirby Siemering)

We found it necessary to remove GFP from the nucleus by localising it to
the ER in order to successfully regenerate bright and healthy Arabidopsis
plants. See Haseloff et al PNAS 1997 (94) 2122-2127 and
http://brindabella.mrc-lmb.cam.ac.uk/
for details.

...........................
Thanks for the replies,
Alvar Carlson
University of Guelph



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