I couldn't find the sequence online, but the lab I work at has a plasmid that holds mKATE, like RFP, that is easily expressible in yeast... (One of the grad students even had his plates infected by yeast with the integration because they spored)
On Sunday, November 10, 2013 2:29:09 AM UTC-8, Mega [Andreas Stuermer] wrote:
-- On Sunday, November 10, 2013 2:29:09 AM UTC-8, 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/ ) fused to the alpha-mating factor which is then cleaved off.6F454BB8401043D798342A07192739 45/Datacards%20or%20Manuals/ N3742Datasheet-Lot0011210.pdf
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.
Should I express it in the cytosol - no golgi etc.?
Thanks,
Andreas
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