Difference between revisions of "Team:Baltimore Bio-Crew/Composite Part"

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               <h2>COMPOSITE PARTS</h2>
 
               <h2>COMPOSITE PARTS</h2>
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          <img src="https://static.igem.org/mediawiki/2017/e/e4/T--Baltimore_Bio-Crew--construct_diagrams.png" style="width: 1000px; height:562px; margin-left:auto;margin-right:auto; display: block;" >  
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We designed two constructs, known as biobricks, to allow the bacteria to express the proteins lipase and esterase that will degrade PET plastics. We designed two constructs, identical except for the sequence for the enzyme each construct would produce: esterase and lipase. Our constructs were designed as follows.
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First is the Lac Regulated Promoter, which starts transcription in the presence of lactose or IPTG. We chose a lac regulated promoter because we wanted to have better control over the expression of the two enzymes.
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Secondly, we had the Ribosome Binding site which signals the start of translation from mRNA to protein.
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During the first year of our project, the design of our constructs was a bit lacking. This year, we made improvements to our biobricks that would allow them to function better. To make sure the  two enzymes  the bacteria produced would be able to exit the cell, we added a PelB periplasmic secretion tag. Once the enzymes were secreted to the periplasm, they would be able to exit the cell on their own.
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After the PelB tag came the gene for plastic degradation. One construct had esterase, and one construct had lipase.
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To allow us to easily isolate the enzymes once they were produced, we added a his tag to our constructs. This would allow us to purify our enzymes on a nickel column before running them on a protein gel. These design changes would contribute to the success of our project by allowing the enzymes to be secreted from the cell and allowing us to easily test for enzyme production.
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Finally, our construct contains a terminator that STOPS transcription of the gene sequence.
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Revision as of 04:21, 1 November 2017



BUGSS Logo
Home Team Project Results Design Notebook Composite Part Safety HP Silver Experiment Attribution Collaborations Interlab Contributions

Baltimore Bio-Crew

BALTIMORE BIO-CREW

Bio-Engineering E.Coli To Degrade Plastic and Save The Baltimore Inner Harbor

COMPOSITE PARTS

We designed two constructs, known as biobricks, to allow the bacteria to express the proteins lipase and esterase that will degrade PET plastics. We designed two constructs, identical except for the sequence for the enzyme each construct would produce: esterase and lipase. Our constructs were designed as follows.
First is the Lac Regulated Promoter, which starts transcription in the presence of lactose or IPTG. We chose a lac regulated promoter because we wanted to have better control over the expression of the two enzymes.
Secondly, we had the Ribosome Binding site which signals the start of translation from mRNA to protein.
During the first year of our project, the design of our constructs was a bit lacking. This year, we made improvements to our biobricks that would allow them to function better. To make sure the two enzymes the bacteria produced would be able to exit the cell, we added a PelB periplasmic secretion tag. Once the enzymes were secreted to the periplasm, they would be able to exit the cell on their own.
After the PelB tag came the gene for plastic degradation. One construct had esterase, and one construct had lipase.
To allow us to easily isolate the enzymes once they were produced, we added a his tag to our constructs. This would allow us to purify our enzymes on a nickel column before running them on a protein gel. These design changes would contribute to the success of our project by allowing the enzymes to be secreted from the cell and allowing us to easily test for enzyme production.
Finally, our construct contains a terminator that STOPS transcription of the gene sequence.