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Team:Nevada/Project
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Contents |
Introduction
Abstract:
Introduction:
Approach
E. coli
Fatty Acid Production
Ethanol Production
-Engineered Pyruvate Decarboxylase and Alcohol Dehydrogenase coding regions (based on Zymomonas mobilis, a microorganism that naturally produces ethanol).
- Removed PDC/ADH from pUC57 vector and put into pSB1C3 -Sequenced and confirmed PDC/ADH in pSB1C3 -Submit PDC/ADH/pSB1C3 to iGEM
-Put a constitutive promoter (σ70) in front of PDC/ADH genes to test expression of ethanol. -Opened up σ70 in Amp vector, then ligated with PDC/ADH. -Sequenced and confirmed σ70/PDC/ADH in Amp vector.
With σ70/PDC/ADH in Amp vector (in NEB 10β cells): -Performed Ethanol Detection test- no samples produced detectable ethanol above background… -Performed ADH Enzymatic test- no ADH detected in assay.
-created “Aldehyde Detection Plates” to test PDC presence & functionality -re-transformed σ70/PDC/ADH in Amp vector into NEB Iq cells -Plates & colonies turn Bright pink- so Aldehydes ARE present (possibly being secreted)
-When ethanol can be detected from σ70/PDC/ADH in Amp vector, I will get -σ70/PDC/ADH into pSB1C3 and submit as an iGEM part. -We are still working on getting pTRC in front of PDC/ADH in pSB1C3
E. coli Results
Cyanobacteria
The cyanobacteria will take up atmospheric CO2 and fix the carbon. From this carbon source sucrose is synthesized, and invertase converts it into glucose and fructose, which is finally transported out of the cell by glF.
