Measurement of Tag Translocation Efficiency in Saccharomyces Cerevisiae Via Confocal Microscopy

Low bioproduction of desired final products is a common problem in synthetic biology. Several technologies exist that can improve bioproduction. Compartmentalization in organelles via signal tagging is one such technology that has been shown to function effectively in Saccharomyces cerevisiae. Compartmentalizing metabolic pathways in organelles can increase bioproduction in several ways, such as improving the proximity of the enzyme to key metabolites, providing a physiochemical environment more ideal for efficient enzyme activity, and increasing the local concentration of metabolic intermediates. Despite its uses in synthetic biology, the translocation efficiencies of tags have never been quantified. This is the goal of the research detailed here. Several tags commonly used in the literature were chosen for the mitochondria (four), the peroxisome (three), and the vacuole (three) and compared with one another. Green fluorescent protein (GFP) was used to visualize translocation under a confocal microscope.