Difference between revisions of "Team:Macquarie Australia/Demonstrate"
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Revision as of 23:37, 23 October 2017
Overview
Key achievements of our team include: Construction and testing of individual parts: hyperlinks to parts registry for Fer, Hyd1, HydEFG, Hyd1/HydEFG and Omega. Improvement of previous part: Hyd1G. Confirmed sequencing of parts. Confirmed transformation into competent cells. Quantifying hydrogen gas production using a Clark electrode. Modelling of hydrogen gas production. Construction of a prototype.
HydG and HydE/F/G
The HydG biobrick was constructed by the 2016 Macquarie iGEM, however it was found to have a 1bp mutation and so lost its functionality. This year we have corrected this mutation and sequenced results to prove we have a functioning maturation enzyme. This biobrick was ligated with biobrick HydEF to assist in the formation of the H-cluster in the Hydrogenase. Confirmed transformation into competent cells (see Fig. 1) and sequencing means this plasmid will allow the faster assembly of the hydrogenase complex, in turn allowing our cell to produce hydrogen gas sooner.
IMAGE HERE
Fer/FNR
Functionality of this biobrick was confirmed this year by running a chromatography to purify proteins with addition of NADH+, and run in a spectrophometer to observe the disappearance of the substrate. Additionally an SDS-PAGe gel was run and bands corresponding to expected band weights were extracted and analysed using MS.
Fer/Hyd
This biobrick was created to ligate a ferredoxin oxidoreductase; an electron transporter via NADP+ reduction, to a hydrogenase native in C. reinhardtii . The ferredoxin oxidoreductase donates electrons to the hydrogenase for the production of hydrogen gas.
