Difference between revisions of "Team:ASIJ Tokyo/Results"

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    <th>Day 0</th>
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    <th>Strong</th>
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    <th>0.05175</th>
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    <th>Moderately-Strong</th>
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    <th>0.071</th>
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    <th>0.07125</th>
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    <th>Weak</th>
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    <th>0.06675</th>
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    <th>0.06825</th>
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    <th>0.06725</th>
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Revision as of 01:59, 18 October 2016

The BIG TEMPLATE : RESPONSIVE and FREE

Results

Overview


The goal of our project was to construct a biobrick capable of plastic degradation. We created three biobrick constructs. Each of these constructs contained PETase, ChloroR, osmY, and an Andersen Promoter. The principal difference between three these constructs was the strength of the Andersen promoter incorporated in the construct. Depending on their predicted ability to promote the secretion of the enzyme PETase by the E. coli bacterium, these Andersen promoters were labelled as weak, moderately-strong, or strong. Our experiment was ineffectual in demonstrating a substantial degradation of PET plastic. The differences in mass measurements of PET after allowing the E. coli bacterium to secrete PETase into the system were insignificant. We believe human errors, such as our failure to feed the E. colicells at appropriate intervals, were a main cause of experimental inaccuracies. In addition, time constraints prevented further data collection. We are unsure regarding whether our PETase construct was successfully secreted into the system. One way we could test this would be to tag PETase with c-myc, and perform a Western Blot test.

Day 0 Day 7 Day 13
Strong 0.05175 0.05175 0.05175
Moderately-Strong 0.071 0.071 0.07125
Weak 0.06675 0.06825 0.06725
graphs




Future Works

Although we were successful in creating a PETase biobrick, we have yet to collect enough data to find an ideal promoter for the construct. As such, the future proceedings of this project would be in gathering a greater amount of data at more frequent intervals in order to identify an ideal promoter.
In addition, we hope to insert an ampicillin resistance gene in our construct, and test for E. coli growth on an ampicillin plate. Testing for another selection factor would act as a secondary confirmation test for the successful construction of a PETase biobrick.
Determining the Andersen promoter with the highest potential to degrade PET plastic would be the first step regarding real-life applications of our lab. The later steps of the PET degradation process, such as the breakdown of Terephthalic Acid and Ethylene Glycol, may be topics of further investigation.