Difference between revisions of "Team:US AFRL CarrollHS/Improve"

Latest revision as of 21:46, 17 October 2018

As part of the gold medal criteria to improve a previous years part, the team decided to improve on British Columbia’s (2013) three parts (BBa_K1129003, BBa_K1129042, and BBa_K1129039) to create the “flavor” of cinnamon through the production of the molecule cinnamaldehyde. These three parts created enzymes to break down the naturally produced, in E. coli., molecule phenylalanine into cinnamaldehyde. Our team decided to take these three separate parts and combine them into a singular part for easier use and cloning. Two ribosomal binding sites were also added in front of the second two parts in order to increase the efficiency of the production of the enzymes to create the cinnamaldehyde (The first part did not need an RBS as the promoter had one attached). In the context of this year’s project, this part would be used in order to produce the cinnamaldehyde molecule as a means of killing off bacteria and fungi that contaminate biofuels in containment systems. This molecule will also kill off the bacteria that is producing it, effectively causing it to be the bacteria’s killswitch. This will ensure that the engineered bacteria will not begin to contaminate the containment system as well. However, due to the part ending up around 5 kb and time constraints, the team was unsuccessful at cloning the part into the iGEM backbone in time for submission and it was not submitted.

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-<div class="row">BBa_K1321340, a double cellulose binding domain with an N-terminal linker, from
+<div class="row"><p>As part of the gold medal criteria to improve a previous years part, the team decided to improve on British Columbia’s (2013) three parts (BBa_K1129003, BBa_K1129042, and BBa_K1129039) to create the “flavor” of cinnamon through the production of the molecule cinnamaldehyde. These three parts created enzymes to break down the naturally produced, in E. coli., molecule phenylalanine into cinnamaldehyde. Our team decided to take these three separate parts and combine them into a singular part for easier use and cloning. Two ribosomal binding sites were also added in front of the second two parts in order to increase the efficiency of the production of the enzymes to create the cinnamaldehyde (The first part did not need an RBS as the promoter had one attached). In the context of this year’s project, this part would be used in order to produce the cinnamaldehyde molecule as a means of killing off bacteria and fungi that contaminate biofuels in containment systems. This molecule will also kill off the bacteria that is producing it, effectively causing it to be the bacteria’s killswitch. This will ensure that the engineered bacteria will not begin to contaminate the containment system as well. However, due to the part ending up around 5 kb and time constraints, the team was unsuccessful at cloning the part into the iGEM backbone in time for submission and it was not submitted.
-Plate 5, Well 18A of the 2017 iGEM Distribution Kit was used by our team to develop a new and beneficial part. The part links the csgA gene from curli fiber DNA to the double cellulose binding domain (dCBD) gene. This new part has been submitted and labelled as BBa_K2522000.</div>
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-<div class="row">Curli fibers are strands of proteins produced by certain strains of E. coli. The fibers are constructed on the outer surface of the cell membrane and encase the bacteria in an extracellular matrix. The genes that make up the DNA of curli fibers are referred to as csg (curli specific gene), and then a letter of the alphabet. Specifically, csgA functions as the building blocks of the hair-like portion of the fibers. The double cellulose binding domain, as the name suggests, attaches to cellulose when cellulose is present.</div>
-<div class="row">To construct the part, the csgA gene was extracted from the genomic DNA of E. coli Nissle and the dCBD was extracted from the iGEM Distribution Kit. The stop codon of the csgA was removed and the two genes were ligated together. In this manner, every time the csgA gene is expressed, a double cellulose binding domain would also be expressed so that the two parts function as a single unit.</div>
-<div class="row">Once a strain of curli-producing E. coli accepts a plasmid containing the csgA-dCBD part, the bacteria can encapsulate itself in cellulose. Such a feature can prove beneficial in various situations. In the context of our project, the cellulose would preserve the engineered microbe from being broken down too early when ingested, or protect the microbe if placed into drinking water. The part could be useful in similar applications where bacteria requires a protective feature. Consequently, developing a plasmid containing csgA-dCBD has improved upon the original function of the double cellulose binding domain part from the Distribution Kit.</div>
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