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Revision as of 13:57, 27 October 2017
Design
Design is the first step in the design-build-test cycle in engineering and synthetic biology. Use this page to describe the process that you used in the design of your parts. You should clearly explain the engineering principles used to design your project.
This page is different to the "Applied Design Award" page. Please see the Applied Design page for more information on how to compete for that award.
Process for simple bioreactor assembly used to measure current produced by Shewanella Oneidensis strains when connected to a potentiostat
Bioreactors
Single chambered bioreactors were designed to be economical and functional. Reactors are run on magnetic stir plates and connected to a lab grade potentiostat.
Plasmid
To produce an electric current from Shewanella oneidensis MR-1 in response to chemicals in a sample, the mtrB gene was first removed from the genome (ΔmtrB) then re-introduced to the bacteria in a plasmid vector under an inducible promoter. The first promoter chosen was the well-studied IPTG inducible T7A1 promoter. Next, nine promoter sequences shown to respond to different effector molecules were identified and amplified from the S. oneidensis MR-1 genome with the addition of the restriction enzyme cut sites for AatII and NdeI on either end. Double digestion of both the vector and promoters using AatII and NdeI removed the T7A1 promoter and produced compatible sticky ends in the new promoter for insertion via ligation.
The designed constructs were individually transformed into E. coli WM3064, sequenced for confirmation, then conjugated into S. oneidensis MR-1 ∆mtrB. The S. oneidensis were then tested for induction through quantification of GFP fluorescence (marker protein) and current production using a spectrophotometer and bioreactor, respectively.
