We are a team of 8 students from Chalmers University of Technology who will represent Gothenburg, SWEDEN in this year’s IGEM competition. We have started with a promising idea that combines the cutting edge technologies available in the field of Synthetic Biology. Our research basically aims to constructing an optical reporter mechanism for cellular stress in yeast by tagging the stress activated SNF1 complex with fluorescent markers.
The project is executed through two main experimental pathways. Both experimental setups will utilize FRET to visualize the conformational change that is the result of the activation of the SNF1 protein. The first approach consists of creating a fusion protein consisting of the SNF1 protein and two fluorescent proteins, namely EYFP and ECFP. The idea is that when the protein is activated it undergoes a conformational change and a FRET-signal will be visible. The second approach utilizes a SAMS-peptide with fluorescent proteins fused to each end. The SAMS-peptide will be phosphorylated by the active SNF1-complex and will undergo a conformational change that will be visible due to the fluorescent tags.
The long term ambition of this project it is to use the results in the pharmaceutical industry when performing high-throughput screening for new substances or finding the correct drug concentrations to use. The yeast cells with the modified SNF-complex can be moved through a micro-fluidic system, gradually exposing them to an array of substances or a concentration gradient and easily finding out at which concentration or substance that the cells are stressed.
As of present we have constructed and ordered primers for all fusion proteins that will be tested if they give a FRET signal in yeast. We are also working on 3D models of the fusion protein and will soon be able to present docking predictions with the complex.
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