Results
![](https://static.igem.org/mediawiki/2016/f/f3/T--ASIJ_Tokyo--ASIJ_fff.png)
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BIOBRICK CONSTRUCTION
Our team was successful in synthesizing a biobrick, as demonstrated by the expression of the Chloro gene when the construct was transformed into E. Coli cultures. Successful growth signified integration of the PETase gene into the plasmid, since the plasmid backbone used (PSIBC3) contained terminator sequences that prevented the expression of Chloro unless the linear backbone was converted into a circular plasmid. The expression of Chloro resistance in E. Coli would thus only have been possible if the assembly of our biobrick had been successful.
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Table 1: Change in Average Mass of PET Film (g) per Promoter
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
Graph 1: Average PET Mass Sample over Time
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.