[DIYbio] Re: Small side project - baking GFP bread - exhibtion

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/6F454BB8401043D798342A0719273945/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