Re: [DIYbio] Re: Engineering Plant Bioluminescence (was Re: experimentation)

I think that comparison is completely bogus, since it assumes that you supply the firefly luciferin!

In fact, if you supply the appropriate luciferin for both luciferases, the bacterial luciferase is infinitely more efficient, since the luciferase reaction uses zero ATP, whereas the firefly luciferase reaction uses one ATP.

Just because we know the biosynthetic pathway for the bacterial luciferin, that should not count *against* the bacterial systems' efficiency! For all we know, perhaps it might take 200 ATP to synthesize the firefly luciferin...

Patrik

On Sunday, March 3, 2013 8:39:16 AM UTC-8, Mega wrote:

> However, maybe, just maybe som leaky expression would be enough...
> 1 Photon needs just one ATP when using firefly luciferase instead
> of 60 ATPs for  bacterial luminescence!

Have you got a citation for that? I was under the impression that
bacterial bioluminescence was in the 90-100% efficiency?

http://mpipz.iwww.mpg.de/25921/koncz_dev_genet_11.pdf

On page "227" (4th page of pdf)

"Comparisons of kinetic parameters and quantum 

yields of luciferases also favoured the firefly enzyme 

which needs only one ATP per emitted photon in con- 

trast to the requirement of 60 ATP for the bacterial 

enzyme "

 

> As long as I get plenty of transformants so that each fragment is
> represented at least once, right?

That's what you need high transformation efficiency to guarantee; to
make sure you have clones of an entire genome, you need huge numbers
of transformants.

On 03/03/2013 09:47 AM, Andreas Sturm wrote:
> Why would I care for transformation effiecncy? As long as I get
> plenty of transformants so that each fragment is represented at
> least once, right?
>
> that you can get insect genes to express in a bacteria, that they
> get the correct posttranslational modification to be active, that
> all the genes for the luciferin biosynthesis pathway are adjacent,
> and close enough together to fit in the bacteria in one piece, and
> that the bacteria has the right metabolic precursors available.
>
>
> Well, it could be that precursors are already present. Sometimes
> living organsms use what is available instead of inventing an
> entirely new metabolic pathway...
>
>
> However, maybe, just maybe som leaky expression would be enough...
> 1 Photon needs just one ATP when using firefly luciferase instead
> of 60 ATPs for  bacterial luminescence!
>
> So if the attached promoter gives just 1/60 of what the pVIB
> promoter gives, it would be brightly visible. If it's less, it
> would still be measurable...
>
>
>
>
>
>
> On Sun, Mar 3, 2013 at 8:38 AM, Nathan McCorkle <nmz...@gmail.com

> <mailto:nmz...@gmail.com>> wrote:
>
> On Sat, Mar 2, 2013 at 9:02 AM, Andreas Sturm

> <masters...@gmail.com <mailto:masters...@gmail.com>>

> wrote:
>> Screening the colonies for activity in this case would be quite
> easy. Turn
>> off the lights, and the ones which don't glow are discarded :D
>>
>
> Unless you want to calculate transformation efficiency, total
> transformants, etc... in that case you need to count ALL bugs, not
> just the interesting ones!!!
>
>>
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