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Carrier on which the biofilm grows
Our biofilm was grow on a carrier material for strength and durability. Whatman paper was chosen for its great biofilm growth and low cost. Four growth methods were studied to optimize biofilm growth and strength.
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Biofilm ion selectivity
The ion selectivity for Na+ and Cl- of a membrane of negatively charged γ-PGA molecules was modelled using Molecular Dynamics. Wetlab testing was performed using B. subtilis Natto.
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Rigidity of the biofilm
To survive water flow, the biofilm has to be stable and robust. The stability was improved by overexpression of biofilm encoding proteins and by knocking out genes having the opposite effect.
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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 locus 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 GMOs for a new project, it is important to know the regulations surrounding genetic modification in general, and think about future perspectives for your project. In addition it is important to realise if and in to what extend you can implement your project and what other people think of this. To address this, we visited COGEM, designed an educational card game and considered several future scenarios involving GMOs and our project.
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Future perspective
To implement our project, the GMOs need to stay in the power plant. Therefore we thought about trapping the bacteria between two carriers, about antimicrobial layers and a kill switch. In addition to not escaping, the ultimate goal is that the biofilm can sustain itself by using nutrients present in the water.