Re: [DIYbio] modified GFP -> pr-coelenterazine peptide

But if the patent is just a hypothesis:

I understood while reading the patent, that on the end there was written that they already did it. At least, it sounded to me as if they would have tried it already with success. Have to read it again.

On Fri, Dec 28, 2012 at 3:09 AM, Jordan Miller <jrdnmlr@gmail.com> wrote:

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

On Dec 27, 2012, at 5:00 PM, Nathan McCorkle <nmz787@gmail.com> wrote:

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 ?

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.

This, however, is a very good point. It truely would be a revolution.

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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