Team:CSU Fort Collins/Results

COLORADO STATE
UNIVERSITY iGEM

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Project Results

Contents

1 Key Results
2 Breakdown
3 Trans-zeatin Production
4 Kill Switch
5 References

Key Results

Key Achievements

• Construction and submission of multiple team parts
• Proof of the composite lac promoter:fadD:fadL part function
• Characterization of KillerRed in E. coli for use as an induced lethality switch

Key Challenges

• Reworking breakdown construct ideas after fadD experiment
• Inconclusive results about trans-zeatin production
• Proper handling and experimental design for testing KillerRed

Breakdown Results

Trans-zeatin Production Results

We ran multiple growth experiments to assess how much metabolic stress, if any, our construct introduced into our strain. From the results, we can conclude that the presence of the genes does not increase metabolic stress on the cell.


Figure 1: Growth comparison between a control strain and our trans-zeatin strain in shake flasks over 24 hours.


Figure 2: Growth comparison between a control strain and our trans-zeatin strain in shake flasks over 72 hours.


Figure 3: Growth comparison between a control strain and our trans-zeatin strain in bioreactors over 72 hours.

Throughout these experiments, we also collected suspension samples which we purified and ran on a reverse-phase HPLC column. Initially, we extracted using 80% methanol and syringe filtration. The HPLC results had too much background to be conclusive, so we developed a new method, based on extraction of zeatin from coconut water^[1]. The procedure included

Kill Switch Results

References

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