Mito Morphin' Power Yeast Project Description

General Overview

As an extension of Team RHIT’s 2015 iGEM project, our goal was to verify the function of MRPS12's mitochondrial localization signal (mls), as well as restore the function of mitochondria in the yeast species Saccharomyces cerevisiae which were deficient in the gene.

Basic Trajectory and Parts

The first portion of the project is to verify the function of the two parts submitted by last year's team. These parts are the mRPS12 translational unit (tu) and the mRPS12 mls. We started by creating yeast vectors that are compatible with the BioBrick standard prefix and suffix. These modified standard vectors facilitated the cloning of our parts into yeast cells and should make working with S. cerevisiae much easier for other iGEM teams in the future. The function of mls was verified by linking it to GFP, cloning it into S. cerevisiae and observing colonies under a florescence microscope. The function of mRPS12 was tested by transforming mRPS12 knock out yeast on two types of complete synthetic media, one made with dextrose and the other made with glycerol, a nonfermentable carbon source. Both types of media were uracil deficient, allowing for the knock out strain to take up the MRPS12 plasmid.