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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 <i>E. coli</i> bacterium, these Andersen promoters were labelled as weak, moderately-strong, or strong. | 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 <i>E. coli</i> bacterium, these Andersen promoters were labelled as weak, moderately-strong, or strong. | ||
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If the team was to carry out this experiment again, perhaps we would gain more significant evidence than we did. We would check for successful expression of our PETase construct, better tend our E. coli, and gather more data over a longer period of time. This data would then help us decide which promoter is best used to optimize the degradation of PET plastic and know that a PETase construct successfully being secreted by the bacteria. Such a conclusion and results could help us determine the next few steps to take in helping our Earth. Of course, we would still have to determine the implications and consequences of using our bioengineered e. Coli and PETase (such as whether or not there would be any acidic waste products, etc.), but knowing which promoter to use to degrade PET plastic more efficiently would be the first step towards applying our lab to the real world. | If the team was to carry out this experiment again, perhaps we would gain more significant evidence than we did. We would check for successful expression of our PETase construct, better tend our E. coli, and gather more data over a longer period of time. This data would then help us decide which promoter is best used to optimize the degradation of PET plastic and know that a PETase construct successfully being secreted by the bacteria. Such a conclusion and results could help us determine the next few steps to take in helping our Earth. Of course, we would still have to determine the implications and consequences of using our bioengineered e. Coli and PETase (such as whether or not there would be any acidic waste products, etc.), but knowing which promoter to use to degrade PET plastic more efficiently would be the first step towards applying our lab to the real world. | ||
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Revision as of 01:24, 18 October 2016
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.
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