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
-- 
Email: echo 36434455860060025978157675027927670979097959886449930P | dc


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