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Team:DTU Denmark/practicalapproach
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The redoxilator - Theoretical background - Yeast as a model organism - Practical approach The USER assembly standard - Principle - Proof of concept - Manual - Primer design software |
The project Practical approach In short here are the milestones of the project: 1. Perform in silico modelling of NAD+/NADH oscillations, Rexivator activity at different NAD+/NADH ratios, reporter gene degradation time, productivity, and more. This should among other things lead to the construction of computer-based model of the sensor system based on NAD+/NADH binding affinity to REX and NAD+/NADH level in yeast. 2. Construct a functional NAD+/NADH sensing system in yeast with a reporter-gene output. All DNA sequences of interest will be chemically synthesized. 3. Run glucose limited fermentation and evaluate oscillative behavior of the reporter expression (quantitatively) in relation to the yeast metabolic cycle. 4. Using NAD+/NADH sensing system to construct a production-strain that produces only in a specific phase of the metabolic cycle (e.g. when NAD+ levels are high). 5. Run glucose limited fermentation, and evaluate oscillative behavior of the productivity (quantitatively) in relation to the yeast metabolic cycle. Compare productivity over time between strains. 6. If necessary, change circuit logic for the production to be in a different phase of the yeast metabolic cycle. 7. Optimize the system by remodeling, based on the previous results. |
Synthetic Biology “Synthetic Biology is an art of engineering new biological systems that don’t exist in nature.” -Paras Chopra & Akhil Kamma In nature, biological molecules work together in complex systems to serve purposes of the cell. In synthetic biology these molecules are used as individual functional units that are combined to form tailored systems exhibiting complex dynamical behaviour. From ‘design specifications’ generated from computational modelling, engineering-based approaches enables the construction of such new specified gene-regulatory networks. The ultimate goal of synthetic biology is to construct systems that gain new functions, and the perspectives of the technology are enormous. It has already been used in several medical projects2 and is predicted to play a major role in biotech-production and environmental aspects. |
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