Green Fluorescent Protein cDNA needed
Dilip at zeus.tamu.edu
Thu Aug 31 11:38:32 EST 1995
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
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.
Texas A & M University
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