Team:Valencia Biocampus/Yeast

THE IDEA

Our aim in this part of the project is to detect when the yeast starts to ferment. At the end of the project we will be capable of “asking” the yeast if there is still any glucose in the media or not by the addition of H2O2. Furthermore we will be able to know how long the media has ran out of glucose. In conclution, this project allows us to know how much time has elipsed since the fermentation began.

To do this, we are going to use two gene constructions:

The ADH2 promoter fused to the YAP1 protein coding sequence. The protein YAP1 is a yeast transcription factor regulator of H2O2 adaptative response. It is stored in the citoplasm in normal conditions and, in presence of H2O2, is transported to the nucleous actting as a transcription factor. The ADH2 promoter is activated in abscence of glucose.

So, complete disappearence of glucose [] the production of YAP1 in the citoplasm whose concentration increases if the lack of glucose continues (we work with delta-yap1 strain).

The TRR promoter is fused to the GFP (Green Fluorescence Protein) coding sequence. The green fluorescent protein can be detected by fluorencent emission. The tiorredoxin reducase promoter is activated by two transcriptional factors (YAP1 and SKN7 in the oxidative form), both only bind to the promoter if H2O2 is previously added to the media.

MOLECULAR MECHANISMS

Click on each plasmid to learn how our constructions work!