it doesn't make evolutionary sense though. GFP was optimized during evolution for fluorescence not coelenterazine. do other coelenterates have GFP-like genes?
and in vivo coelenterazine synthesis has not been figured out AFAIK.
the patent is a hypothesis, not a proof of concept or reduction to practice.
jordan
Seems that this whole patent-not-working points to the coelenterazine not spontaneously forming from the modGFP alone. I bet there are some other enzymes in there that recycle/recharge the modGFP
On Dec 27, 2012 11:38 AM, "Andreas Sturm" <masterstorm123@gmail.com> wrote:--On Thu, Dec 27, 2012 at 7:49 PM, Jordan Miller <jrdnmlr@gmail.com> wrote:
we did the point mutation on eGFP optimized for bacterial expression with the induced vs. non-induced controls and coelenterazine as the positive control with purified Gaussia luciferase from a mammalian expression system. did not work as expected in our hands.
Ok, but the sequence of eGFP differs from natural wild-type GFP... You just changed Amino acid 65 Ser -> Tyr ?This, however, is a very good point. It truely would be a revolution.the ability to get this would revolutionize non invasive imaging since you wouldn't need to inject any luciferin/coelenterazine, so of it did work it would have been a big paper by the inventors. the most worrisome thing to me is they appear to have dropped the project.
But I had had a research some times ago on how much a patent costs, and it's horrible. And, there's no world-wide patent, you have to buy it for each country seperately. So you hardly get a patent in most of the countries (US, some European countries, Brazil, Russia) below 10'000 $ !
Why would they throw away > 10k $ for patenting something that doesn't work??
Did you get really *no* luminescence? Had a HPLC on it?
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