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From 2012.igem.org

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Overall project

Vitamin Deficiency is a health problem that is prevalent throughout the world. The particular problem that we have decided to focus on is the widespread problem of vitamin D deficiency. There are two kinds of vitamin D: D2 and D3. Each of these comes from a different source and can be utilized by the human body, however evidence has shown that vitamin D3 can be used more effectively by the human body. While this vitamin can be synthesized by the human body, this is only capable of occurring during certain months of the year and if the person chooses to expose their bear skin to the sun for the amount of time. Our solution the this problem is to create a baker's yeast strain, S. Cerevisiae, that can synthesis vitamin D3. This yeast strain will be able to, in theory, be baked into foods and used in the fermentation of drinks thereby infusing these items with vitamin D3.

Project Details

Our iGEM project was to create a yeast strain that is able to synthesize vitamin D3 into a food. To do this we identified the 2 genes that are not prevalent in yeast that are needed to produce the proteins needed to produce the precursor to D3 in yeast. having Identified these gene was designed primers to PCR these genes from Human cDNA. Alas, due to time and budget constraints, we were unable to PCR these genes, so we ordered them from GeneCopia. These genes came in a Gateway entry vector. having ordered this part we used the Gateway vector system from Invitrogen. This system allowed us to easily grow the entry vector in E. Coli and transfer the genes of interest into the destination vector, pYES-DEST52. This was done using the Gateway LR Clonase Enzyme Mix. We are now in the process of transforming the yeast with these two vectors. The problem that we are facing is that we are playing a game of chance. The two vectors with the different genes have the same selection markers. When transforming the yeast cell, we are hoping that both vectors will go into the yeast. However we will not know this until we test for expression of the genes in the yeast.

If we had more time to properly PCR the genes we would have liked to put these into two separate vectors, (find bens vector names) . This would have allowed us to have two selectable markers for the genes and the only thing we would have left to do is confirm that the proteins are making vitamin D3 in yeast.

Results