GFP in plants

Xavier Gansel Xavier.Gansel at unifr.ch
Wed Jan 29 13:30:50 EST 1997


Dear netters,

A few weeks ago I posted the following question to this news-group as well
as to people known, from the bibliography, to be working on GFP in plants.

>Here is a mail to ask for advices on the use of GFP in A. thaliana
>I wish to use GFP as a reporter gene to follow the regulation
>(activation/repression, tissue and cell expression, etc. ....) of a
>promoter.
>So, I want to get transgenic A.t. plants carrying a GFP under the control
>of this promoter.
>
>From the bibliography, it appears that their is a growing number of "plant
>optimised" GFP. The optimisations beeing on different aspects. Some of them
>are the optimisation of codon usage, the removal of a cryptic A.t. intron,
>the addition of a plant intron, mutations that shift light emission or
>enhance thermoresistance, anchoring in the ER to reduce toxicity of nuclear
>GFP accmlulation (Chui, 1996, Curr. Biol., 6, 325-330 //  Davis, Weeds
>World Volume 3(ii) // Haseloff, TIG, 1951, 11, 328-329 // Haseloff, poster
>S44, 7th International Conference on Arabidopsis Research // Pang, Plant
>Physiol., 1996, 112, 893-900).
>But all those mutations/improvements are not present on the same plant-GFP.
>
>My questions are the following :
>1/ on today statut, which version of plant improved GFP would you recommend
>to be use as the best reporter system in A.t. ? (the promoter to be studied
>may be weak and so require a very sensitive reporter gene to detect it)
>2/ were could I get this "super" plant-GFP ?

Up to today I obtained only the two following answers. Of course I thank a
lot Jim Haseloff and Seth J. Davis for the kind informations they provided.

Cheers,

Xavier.
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1//
From: sjdavis1 at students.wisc.edu (Seth J. Davis)

Dear Xavier Gansel,

1)  What is the best GFP for promoter studies in Arabidopsis thaliana?
2)  Where do you get it?

The first question is difficult to answer, as none of the plant-use GFPs
have been fully characterized.  In Arabidopsis I believe several factors
should be included in any choice.  First, the optimized codon use should be
included, and a soluble derivative should be used.   Under this criteria,
there are GFPs generated from two labs: the Vierstra Lab and the Haseloff
Lab.  It is not clear who's GFP is "better."  But if your going to use the
GFP for promoter studies, and have the capacity for fluorescensce
microscopy,  I would choose either psmRSGFP or mGFP5.  These two GFPs have
not been compared sized by side in promoter studies, but it is being done
here at the UW-Madison (by several labs).

As for the second question, psmRSGFP can be obtained from the Arabidopsis
Stock Center at the Ohio State University.  Haseloff's email is
jph at alf1.mrc-lmb.cam.ac.uk.  You should contact him for mGFP5.

Seth
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2//

From: jph at mrc-lmb.cam.ac.uk (Jim Haseloff)

Hi Xavier,
                I'll have you sent some of our latest modified GFP - it
comes with full details, and should be want you want - There are partial
details in the December issue of Current Biology.
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