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GFP variants

Madeleine Rashbrooke RASHBROOKE at rsbs.anu.edu.au
Thu Mar 15 02:04:21 EST 2001


Dear All,

I am in the process of planning a project involving the construction of a
fusion of a fluorescent protein to my protein of interest. In order to
leave
open the possibility of double labelling with existing GFP constructs, I
need to use a non green variant. I have a couple of questions that I was
hoping somebody would be able to help me with, based on their own
experience
with fluorescent tags in arabidopsis.

Firstly, what is the best source of arabidopsis-optimised fluorescent
protein coding sequences? Clontech have an extensive collection
advertised,
and claim that they work well in plants, but has anyone first hand
experience of using these vectors for arabidopsis projects?

My other question relates to pairs of fluorescent protein colours - I
gather that cyan and yellow are the best combination, especially if one
wishes to try FRET, but has anybody had any success with double labelling
of
GFP and the new "red" variants such as Clontech's dsRED? Alternatively is
it
possible to separate the spectra of cyan and green, and green and yellow,
sufficiently to label two different proteins in the same cell with these
pairs?

I promise to post any (or all) replies back to the newsgroup - I suspect
it's not just me who's confused....

Thank you very much in advance,

Madeleine


Madeleine Rashbrooke
PhD student, Wasteneys lab
Plant Cell Biology Group
Research School of Biological Sciences
The Australian National University
GPO Box 475
Canberra ACT 2601
Tel: +61-2-6125 5561
Fax: +61-2-6125 4331
Email: rashbrooke at rsbs.anu.edu.au

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