Green Fluorescent Protein cDNA needed

DILIP DIAS Dilip at zeus.tamu.edu
Thu Aug 31 11:38:32 EST 1995


Yves:
There are two sources for GFP cDNA. 
1. Clonetech, USA. The GFP available from them is the one that was 
isolated by the group in Columbia University in New York.  This works in 
plant systems but sometimes there is cryptic splicing that occurs in 
plants that render the GFP inactive (The GFP is rich in A & T and mimic 
an intron which is removed by plant systems).
2. The problem of cryptic splicing has been removed by a group in 
Edinburgh, U.K. (See J. of Cell Biol. 121:83-90) and is reported to be 
better in plant systems. They have modified the GFP to increase the GC 
percentage. Contact them and they have been gracious to give the cDNA 
free already in pBIN vector ready to transform the plants. Clonetech 
cDNA is not in pBIN vector.

There are certain things that you should be aware of GFP in plant 
systems.
1. Lignin autoflouresces at the same excitation wavelength as GFP and 
will give green color emission under fluorescent microscope even in 
untransformed plants.  If the promoters you are going to study are 
associated with vascular system forget about GFP.
2. GFP is not an enzyme. Therefore the signal you get is not amplified 
as in GUS.  Hence, if your promoters are weak then you mightnot see any 
signal at all.
I hope these will help you.
Dilip Dias
Texas A & M University
Texas





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