Dear GFPers
A while ago I put out a request for information on what GFPs people are
using for reporter constructs. At long last I am getting the summary of
responses out to the group. I would like to thank those people who took
the time to let me know what they are doing with GFP. I really learned a
lot!
I scored the responses for three things: promoter type, codon usage, and
GFP mutant employed. Some responses did not contain information about all
three categories. I also included whatever I could find on various
companies Web sites as "responses".
PROMOTERS
About half were using strong constitutive promoters like CMV, CaMV or
actin. Others were studying inducible or virallly promoted constructs.
My personal favorite was transgenic mouse lymphocytes expressing GFP from
the IL2 promoter in response to mitogen stimulation.
CODON USAGE
humanized: 7/13
plant splice optimised: 4/13
A. victoria: 2/13
GFP MUTANTS
Mutations fall into two categories. Some like S65T affect primarily
fluorophore exitation and emmission properties. Others like GFPuv affect
expession, folding kinetics and solubilty of the GFP protein as a whole.
Of course these categories are not truly mutually exclusive. Some labs
have combined solubility enhancing mutants with fluorophore mutants to
reach a further level of optimisation.
S65T 38%
EGFP 19%
Wild-type 9%
RS-GFP 9%
Y66H 9%
GFPuv 14%
Other 2%
Hope that this info will be useful to othersas it was to me.
MIKE
Mike Moser Tel: 206-616-7391
UW Department of Pathology FAX: 206-543-3644
Box 357470 moser at u.washington.edu
Seattle, WA 98195 http://weber.u.washington.edu/~moser
