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| - | <p>Malnourishment has been a problem in the world for as long as history has been recorded. Many of the complex sugars found in plants cannot be broken down by humans or even the ruminants of cattle or goats. By breaking these complex sugars down into digestible sugars, the caloric value of these foods could be increased. Through these means we hope to make a large stride toward lessening the world’s caloric deficit. </p><p> | + | <h1><center>Sugar Rush</center></h1> |
| - | Three enzymes we’re isolated from <i>Bacillus subtilis subtilis 168</i> for use in this project: yesZ, xynA, and bglS. All three of these enzymes play an individual role in the breakdown of biomatter in a digestive system. YesZ cleaves RG I Pectin, which is a primary component in the cell wall of plants. Pectin is cleaved into single galactose molecules that can be metabolized by both human and animal digestive systems. XynA degrades xylan, a polysaccharide chain that cannot be broken down by the human body. Xylan is a major structural component of plant cell walls, and, due to crosslinks with cellulose and other cell wall components, inhibits the access of cellulases. BglS is a beta-glucanase that breaks down beta-glucan. BglS breaks glycosidic bonds within beta-glucan and hydrolyzes these glucans making the substance more easily digestible.</p><p>
| + | <h2><center>Wisconsin Lutheran College - Milwaukee</center></h2> |
| - | In addition to caloric deficit, norovirus outbreaks frequent our project’s target populations. The lower quality of living and increased water contamination allows easy transmission throughout these developing nations. Infected individuals experience gastroenteritis and severe dehydration, at worse, leading to death.</ p><p> | + | <p> |
| - | Our plasmid includes an antinorovirus particle antibody in fusion with LPP-OmpA. Once expressed, the antinorovirus monocolonal antibody binds to the C-terminus of the chimeric protein LPP-OmpA. The protein introduces itself into the outer membrane, thus displaying the antibody extracellularly. The antibody then binds to native, recombinant norovirus particles. This inhibits the virus’s interaction with the infected individual's cells, which ultimately allows the organism that ingested the probiotic to successfully combat the virus.</ p><p> | + | The goal of our research project is to engineer a probiotic bacterium to secrete the cellulase enzyme that will break down indigestible cellulose in plant material to its digestible glucose components. By introducing this bacterium into an animal, the animal would be able to gain nutritional sustenance from plant material that is normally passed as waste. Molecular biology methods will be performed to clone a cellulase gene into a cloning vector that will allow this enzyme to be produced in a bacterial cell and ultimately secreted into its environment. Specific genes will also be incorporated into the bacterial strain to ensure the new cellulase-secreting strain will not be harmful to the environment. |
| - | Our final plasmid has a portion dedicated to anti-horizontal transfer, preventing the DNA from escaping into the wild or being misused. This section, referred to as “Safety,” is compromised of the Tse2 intracellular toxin gene (BBa_K314200) downstream of a T5 Cumate Operator (BBa_K875001). The molecule CymR, which is natively expressed in E. coli Nissle 1917, represses this operator. Bacteria that are transformed by this plasmid that will manufacture the toxin and self-destruct. In addition, the CymR repressor can be inactivated by another molecule, p-cumate. The combination of these two properties allows our DNA to be used in a controlled fashion, allowing for cheap treatment of both intentional or unintentional outbreaks.</p><p>
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| - | The target countries of our project hold cultural practices that differ greatly from our own. Because of this, our team strives to be culturally sensitive and uphold the cultural practices of the indigenous people. Others that have attempted to help these natives have unknowingly invaded the traditional cultural practices. While these attempts did increase the natives’ quality of life, they often times, destroyed the portion of their lives that defined the culture. We are attempting to introduce a product that doesn’t interfere with the indigenous culture, while simultaneously positively affecting their quality of life. Our project will allow farmers to tend to their herds, but allow them to use fewer resources to sustain them.</p><p>
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| - | We have also investigated the key role education plays in effectively improving quality of life of others. Education is vital in the assistance of different countries. This includes the education of the indigenous individuals, but also the non-native peoples. Our team took it upon ourselves to help educate future scientists in the subject of biotechnology by hosting a week long summer camp for high school students. During this camp we focused on introducing students to the basics of cellular biology and biotechnology’s potential to solve grand world problems. </p>
| + | Our team also aimed to assess feelings towards biotechnology through a survey that we administered. We reached out to our community by hosting a camp for high school students. Our camp served home schooled students and students from area Lutheran high schools. We provided these students with the opportunity to gain lab experience which they likely would not have had access to. |
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| | + | <h3>Sugar Rush Abstract</h3> |
| | + | <p>In parts of the world suitable land to raise grazing animals is limited so these animals are either not raised or are malnourished. Animals can get more nourishment from a smaller amount of land if they increase digestion efficiency by converting more sugar polymers (cellulose) from plants into usable sugars. We aim to construct an <i>Escherichia coli</i> strain capable of secreting cellulose-degrading enzymes in an animal’s gut. This strain will contain a plasmid that expresses the bglS, yesZ, and xynA genes from <i>Bacillus subtilis</i> that code for three enzymes that cleave different bonds in cellulose. These enzymes will be secreted from the well-studied probiotic <i>Escherichia coli</i> Nissle 1917 cell by a Type I secretion system whose genes are also engineered into the plasmid. In raising animals with this engineered bacterium, we hope to increase the amount of available calories for nutrient-deprived populations while having minimal effects on established cultural practices.</p> |
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