Team:DTU Denmark/results

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The project

Notebook

The redoxilator

- Genetic design
- Applications and perspectives
- Results
- Safety considerations

The USER assembly standard

- USER fusion of biobricks

USER fusion primer design software

- Abstract
- Instructions
- Output format

The project

Results

Two strains were grown under diferent substrates, Glucose, Galactose, Glucose and Galactose and Ethanol. These two strains that we denominate Construct and Leaky correspond to the redoxilator with a constitutive promoter and a minimal promoter repectivelly.

Growth

Growth rates (h-1) obtained for the two strains on diferent carbon sources

Carbon Source	Leaky	Construct
Glucose	0,114 ± 0,006	0,112 ± 0,003
Galactose	0,084 ± 0,010	0,076 ± 0,003
Ethanol	E0,013 ± 0,005	0,007 ± 0,001

As expected, both strains showed different profiles of growth accordingly to the substrate. For Glucose and Galactose, the growth curve showed two exponential phases, the first one corresponding to the growth on Glucose and the second corresponding to the growth on Galactose. Although the initial OD was similar in both cases, the growth on Galactose showed an even longer lag phase than the glucose. Finally, as expected, the substrate reporting less growth was ethanol. For the Construct, the growth solely on Glucose showed a longer lag phase compared to the mixed substrate, probably due to a lower initial OD.

File:OD Construct.jpn

Growth profile (OD) of the Construct for the different carbon sources

File:OD Leaky.jpn

Growth profile (OD) of the Leaky for the different carbon sources

Achievements

Redox sensing device
Two novel genes have been designed and synthesized each comprised of 5 genetic elements. Together they function as a device termed the Redoxilator that can sense the internal redox state of a yeast cell, and output a reporter signal.

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

We have demonstrated that our USER fusion biobrick works as expected and documented it (silver medal)

USER fusion Assembly standard
A new biobrick assembly standard that allows the rapid construction of multi-part devices have been developed and documented. 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)

USER-fusion primer design software
A novel and very useful software tool have been developed that can automatically design the optimal primers for USER fusion assembly of 2-9 biobricks, taking several parameters into account.

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