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Carrier on which the biofilm grows
The biofilm had to grow on a carrier material for strength and usability in the prototype. Whatman paper was chosen for its great biofilm growth and low cost. Four growth methods were studied to really optimize biofilm growth and strength.
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Biofilm ion selectivity
The J-PGA molecule was likely to improve the energy that could be generated with the test set up.. Therefore we made several models with Molecular Dynamics after this did tests in the lab with natto strain of Bacillus subtilis.
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Rigidity of the biofilm
To survive the waterflow the biofilm had to be stable and robust. Biofilm genes have been overexpressed to be constantly transcribed. Furthermore stability has been improved by repressing and knocking out genetic pathways resulting in cell motility and biofilm repression.
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New shuttle vector
The BBa_K823023 backbone is one of the standard backbones for bacillus for integrating in the amyE locus. However an extra integration locus is welcome when making a multiple mutant. Another integration loci is the thrC locus. To create this backbone the BBa_K823023 backbone was used, where the amyE locus parts were replaced with the thrC locus parts from the plasmid pdg1664. This resulted in a new shuttle vector for B. subtilis.
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Human practices
When working with GMO’s it is important to know the regulations and think about future perspectives, which we did with possible future scenarios. Also it is important to talk about iGEM and your project at events, with several media, at this occasions we used our own card game.
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Future perspective
To implement our project the GMOs need to stay in the power plant. Therefore we thought about capsulating the bacteria into two carriers, antimicrobial layers and a kill switch.The ultimate goal is that the biofilm can sustain itself from nutrients in the water.