Questions about protein quantification - from the industry
perspective
Aawara Chowdhury
via methods%40net.bio.net
(by aawara from pontiff-playground.org)
Tue May 27 06:48:09 EST 2008
In <g1ecv3$ukc$1 from kuling.itea.ntnu.no>,
Trond Erik Vee Aune <trondaun from REMOVE.nt.ntnu.no> wrote:
> What I need is a tag to be used in our own expression vectors for in
> vivo quantification of gene expression. I was thinking about GFP but I
> am afraid that commercial use of this protein is patented. So,
> disregarding GFP, what other cheap alternatives are there? It might
> easily be so that licensing GFP for our use is the least expensive
> alternative, but I won't go for this solution before giving it a lot of
> thought ;).
>
> I've also heard that it is not straight-forwarding using GFP for
> quantification of expression. Anyone has and thought or experiences on this?
GFP is an extraordinarily stable protein - which may compound your
analysis.
How about using beta-lactamase? Here's the original reference from
Roger Tsien's group - <http://www.ncbi.nlm.nih.gov/pubmed/9417030>
Since that publication, others have used beta-lactamase fusions to
measure gene expression in live mammalian cells.
<http://www.ncbi.nlm.nih.gov/pubmed/11036645>
And now, there are reagents to do this via flow cytometry ...
<http://www.ncbi.nlm.nih.gov/pubmed/12541281>
Cheers,
AC
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