Team:DTU Denmark

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Sponsors and acknowledgements

- List of sponsors
- Acknowledgement

DTU Denmark


Project abstract

The Redoxilator

By in silico design and computer modelling followed by gene synthesis, we have constructed a molecular NAD/NADH ratio sensing system in Saccharomyces cerevisiae. The sensor works as an inducible transcription factor being active only at certain levels of the NAD/NADH ratios. By the coupling of a yeast optimized fast degradable GFP, the system can be used for in vivo monitoring of NAD/NADH redox poise. A future novel application of the system is heterologous redox coupled protein production in yeast.

The redox coupled system

The USER fusion standard

Another part of our project is the proposal of a new parts-assembly standard for Biobricks based on USER(TM) cloning. With this technique, not based on restriction enzymes, all parts independent of function can be assembled without leaving any scars from the restriction enzyme digestions.


The USERTM fusion primer design software
Design your primers here

ACHIEVEMENTS
We have...
1) Designed and synthesized two novel genes each comprised of 5 genetic elements. The constructed device is termed the Redoxilator, and can sense the internal redox state of a yeast cell, and output a reporter signal.

2) Designed and submitted DNA of several new biobricks including a yeast optimized GFP reporter protein, a protein degradation sequence and a fast degradable yeast GFP. (Bronze medal)

3) Demonstrated that our USER fusion biobrick works as expected and documented it (silver medal)

4) Developed and documented a new biobrick assembly standard that allows the rapid construction of multi-part devices. The assembly standard offers many benefits: All restriction sites are allowed, multiple biobricks can be joined in one step, the result is scar-free making it ideal for protein fusions and more. (Gold medal)

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