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> wrote:
On Sat, Mar 2, 2013 at 9:02 AM, Andreas Sturm <masterstorm123@gmail.com> wrote:Unless you want to calculate transformation efficiency, total
> 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
>
transformants, etc... in that case you need to count ALL bugs, not
just the interesting ones!!!
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