promoter GFP-GUS fusion

Alan Smith smitam01 at holmes.ipfw.edu
Tue May 16 12:08:09 EST 2000



Mat wrote:

> Hi all,
>
> I isolated a bit of a promoter and would like to fuse in with GFP-GUS
>
> 1) Do you need to keep a bit of the 5'UTR along with the promoter or
> will the promoter part alone will do fine?
>

I would make the promoter constructs contain the 5' UTR just to be safe.
You may to make a few different constructs....constructs with the UTRs,
constructs with out UTRs, constructs with leader peptides, deletion
constructs, etc........Just to be safe.  I have read many plant promoter
papers and most papers that I have read have the 5'UTR in the promoter
construct.  This may be different in other organisms.


>
> 2) I have a plasmid with GFP-GUS construct. Anybody has tried to fuse a
> promoter in front of them and manage to detect GUS and GFP "activity " ?
>

I have never used GFP-GUS fusions before, but check out this paper
Quaedvlieg N. et al.  Fusions between green fluorescent protein and
B-glucuronidase as sensitive and vital bifunctional reporters in plants.
Plant Molecular Biology  37:715-727. 1998.


Alan Smith



>
> Advices are welcome.....
>
> Thank you
>
> Mathieu
>
> ---

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