Team:Valencia Biocampus/Yeast

From 2012.igem.org

Yeast Subteam

The Idea

Our aim in this part of the project is to detect when the yeast starts to ferment. To do this, we are going to use two gene constructions. The first construction consists of the ADH2 promoter fused to the YAP1 protein coding sequence. In the second construction the TRR promoter is fused to the GFP (Green Fluorescence Protein) coding sequence. We use the ADH2 promoter (alcohol dehydrogenase) to sense the glucose/ethanol concentration. When this promoter is activated, YAP1 is expressed. This protein will attach to TRR promoter (thioredoxin reductase), which will become activated if the constitutive yeast protein SKN7 is in its oxidized form and also interacts with the promoter. This activation will enable transcription of GFP. H2O2 has to be added in order to oxidize the SKN7 protein.

Protocols

Here goes the protocols