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

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Wow, I had no idea it was so terrible! How can there be such a vast
gulf in efficiency?

On 03/03/2013 04:39 PM, Andreas Sturm 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
>> <nmz787@gmail.com
> <mailto:nmz787@gmail.com>
>> <mailto:nmz787@gmail.com <mailto:nmz787@gmail.com>>> wrote:
>>
>> On Sat, Mar 2, 2013 at 9:02 AM, Andreas Sturm
>> <masterstorm123@gmail.com <mailto:masterstorm123@gmail.com>
> <mailto:masterstorm123@gmail.com
> <mailto:masterstorm123@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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