Team:UrbanTundra Edmonton/Proof
- TEAM
- PROJECT
- PARTS
- SAFETY
- ATTRIBUTIONS
- HUMAN PRACTICES
- ACHIEVEMENTS
The Bio Reaction
Aminolevulinic Acid and IPTG
The LacI gene in our expression plasmid codes for Lac repressor proteins that bind to Lac operators flanking the T5 promoter. IPTG was added to our E.coli cultures prior Oxygen production in order bind to the Lac repressors, releasing their bind on the Lac operators. This in turn was allowed the expression of the Chlorite Dismutase gene since RNA polymerase was then able to bind to the T5 promoter, initiating the transcription process. The Chlorite Dismutase enzyme has a heme component in its protein structure. Aminolevulinic acid, a heme precursor, and ferric sulfate which provided the iron needed for heme production, were added to the E. coli cultures to allow for proper synthesis of Chorlite Dismutase.
Oxygen Production
Having successfully transformed the Cld- gene into our E. coli chassis, we progressed to test the capability of the Chlorite Dismutase enzyme to convert chlorite ions (ClO₂⁻) into the useful byproduct, oxygen gas (O₂) and chloride ions in solution (Cl-). The purpose of this investigation was to determine whether or not the enzyme synthesized by the chassis was functional. Three trials of this investigation were conducted, where solid Sodium Chlorite (NaClO₂), at concentrations of 0.1M and 0.2M, were introduced into two separate volumes of transformed E.coli cultures suspended in 50 mL of LB Broth. The system was closed immediately after the addition of the enzyme’s chlorite substrate and the oxygen produced by enzymatic action was captured with a specifically allocated balloon which the height of was measured using a ruler. The results of this investigation will determine if our construct does work properly and roughly indicate the experimental yield to expected.
Concept Sketch
Explore With Us.
