Team:StanfordBrownSpelman/Biodegradability

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Stanford–Brown–Spelman iGEM 2014 — Amberless Hell Cell Hello World

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Results
Our goal for this project was not only to isolate biodegradation enzymes but also to control the release of these enzymes, so that our UAV would not degrade uncontrollably. In order to control the initiation of biodegradation, we first considered using a pressure sensor. This would allow the UAV to begin degrading after the impact of a crash. The 2008 Tokyo Tech iGem team found that the ptet promoter was pressure sensitive and increased its activity 3-fold after undergoing 30 MPa of pressure. However, after analyzing the impact a crash would have on our UAV by using a force plate, we found it unlikely that the impact of the crash would ever reach such a high pressure (see graph below). Rather than searching for another type of pressure sensor, we chose to try another method. '''JEANETTE OR JOVITA PLEASE INSERT THE PRESSURE/FORCE PLATE GRAPH HERE. IT IS NOT IN THE GOOGLE DRIVE SO I CAN'T DO IT.''' We next tried to initiate degradation using a light sensor, which would activate degradation in the darkness, allowing our UAV to have a flight time of one day. We planned to do this using the construct from the UT Austin and UCSF 2004 Coliroid project. However, the strain of E. coli (EnvZ) that we needed to work with to use this construct was resistant to all 4 of the main antibiotics we had in our lab, making the bacteria difficult and expensive to work with. Finally, we decided to use quorum sensing as a means of creating a time delay for initiation of degradation. Two previous constructs, BBa_I13202 and BBa_T9002, when combined, create a quorum sensing cascade used to initiate expression of GFP. We ligated these two parts together to create our novel part, BBa_K1499500, which we used for our assays. We found that, in lac deficient cells, the quorum sensing construct can be initiated by induction with IPTG. After this induction, GFP expression increases with time, implying that the construct is doing its job. Our data for this assay is shown below. '''ARYO PLEASE INSERT THE FLOW CYTOMETRY DATA HERE SINCE I DON'T HAVE IT.''' Since we know that the quorum sensing construct is functional and inducible with IPTG. We can now work towards replacing the GFP gene with the genes for our degradation enzymes, allowing us to control degradation by applying IPTG at different time points.
Methods & Safety
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Links & References
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Additional Information
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