I'm more concerned that there won't be time during rising for yeast to make enough GFP: have you considered a sourdough instead? Coculture bacteria and yeast, long fermentatiob, plenty of time for GFP?
"Mega [Andreas Stuermer]" <masterstorm123@gmail.com> wrote:
P.S. I have considered that baking will destroy GFP anyway. But the raw dough will make beautiful fotos :D
On Sunday, November 10, 2013 11:29:09 AM UTC+1, Mega [Andreas Stuermer] wrote:Hi everyone,
I am condsidering as a "weekend project" to add GFP to a yeast (saccharomyces, considered GRAS as you know), and then bake bread with it. Haven't talked to the guys from Ars Electronica yet if their Biolab would host this (but I'm positive, if it is legal they will allow it).
Therefore I would subclone GFP into pKlac2 ( https://www.neb.com/~/media/Catalog/All-Products/ 6F454BB8401043D798342A07192739 45/Datacards%20or%20Manuals/ N3742Datasheet-Lot0011210.pdf ) fused to the alpha-mating factor which is then cleaved off.
The GFP would then be secreted into the medium (bread dough). So if you cut the dough, there will be small holes which will be fluorescent.
What is questiuonable, will the GFP still work? If it is secreted the yeast will glycosylate it, unlike most cytosolic proteins (according to http://wolfson.huji.ac.il/purification/PDF/Expression_ Systems/NEB_Klactiskit_manual. pdf )
Should I express it in the cytosol - no golgi etc.?
Thanks,
Andreas
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