Difference between revisions of "Team:Austin UTexas/Demonstrate"

 
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<h2>Demonstrate</h2>
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<h2> Demonstrate </h2>
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<br>
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<p>For the gold medal “Demonstrate your work” requirement, we have shown that we can recreate kombucha from scratch by adding isolated strains of microbes to tea media. Our tea media, made with water, black tea, and sucrose, simulates the starting mixture used to brew kombucha. We have performed extensive recapitulation trials to determine which strains of yeast and bacteria are necessary for brewing kombucha. Cataloguing these vital strains is a necessary step toward modifying the starting population to create a “designer beverage” with a variety of properties that could benefit kombucha consumers and manufacturers outside the lab. </p>
−
<p> Click on one of the images below to learn more about our results! </p>
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<p>Click the images below to learn more about our results!</p>
 
</div>
 
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<input type="image" src="https://static.igem.org/mediawiki/2016/4/40/T--Austin_UTexas--StrainNavi.png" style="width:100%"; onclick="showOne('section1')"/> <p>Kombucha Strains </p>
 
<input type="image" src="https://static.igem.org/mediawiki/2016/4/40/T--Austin_UTexas--StrainNavi.png" style="width:100%"; onclick="showOne('section1')"/> <p>Kombucha Strains </p>
 
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<input type="image" src="https://static.igem.org/mediawiki/2016/6/64/T--Austin_UTexas--ConjugationPic.png" style="width:100%;" onclick="showOne('section2')" /><p>Conjugation </p>
 
<input type="image" src="https://static.igem.org/mediawiki/2016/6/64/T--Austin_UTexas--ConjugationPic.png" style="width:100%;" onclick="showOne('section2')" /><p>Conjugation </p>
 
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<input type="image" src="https://static.igem.org/mediawiki/2016/0/04/T--Austin_UTexas--RecapNavi.png" style="width:100%;" onclick="showOne('section3')" /><p>Recapitulation</p>
 
<input type="image" src="https://static.igem.org/mediawiki/2016/0/04/T--Austin_UTexas--RecapNavi.png" style="width:100%;" onclick="showOne('section3')" /><p>Recapitulation</p>
 
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<input type="image" src="https://static.igem.org/mediawiki/2016/b/bb/T--Austin_UTexas--EtOHNavi.png" style="width:100%;" onclick="showOne('section4')" /><p>Ethanol</p>
 
<input type="image" src="https://static.igem.org/mediawiki/2016/b/bb/T--Austin_UTexas--EtOHNavi.png" style="width:100%;" onclick="showOne('section4')" /><p>Ethanol</p>
 
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<input type="image" src ="https://static.igem.org/mediawiki/2016/e/e7/T--Austin_UTexas--pHNavi.png" style="width:100%;" onclick="showOne('section6')" /><p>pH</p>
 
<input type="image" src ="https://static.igem.org/mediawiki/2016/e/e7/T--Austin_UTexas--pHNavi.png" style="width:100%;" onclick="showOne('section6')" /><p>pH</p>
 
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<input type="image" src ="https://static.igem.org/mediawiki/2016/9/92/T--Austin_UTexas--GellanNavi.png" style="width:100%;" onclick="showOne('section7')" /><p>Gellan Gum</p>
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*Identified strains of bacteria and yeast using rRNA gene sequencing.
 
*Identified strains of bacteria and yeast using rRNA gene sequencing.
 
*Characterized each of the isolated microbes to facilitate further experimentation.  
 
*Characterized each of the isolated microbes to facilitate further experimentation.  
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NEED LINK
 
  
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<a href ="https://2016.igem.org/Team:Austin_UTexas/Results#section1">Results</a> (May need to open in a new tab.)
 
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*Performed minimum inhibitory concentration experiments between <i>G. oxydans</i> and spectinomycin, carbenicillin and kanamycin.
 
*Performed minimum inhibitory concentration experiments between <i>G. oxydans</i> and spectinomycin, carbenicillin and kanamycin.
 
*Determined that <i>G. oxydans</i> is resistant to spectinomycin and carbenicillin.
 
*Determined that <i>G. oxydans</i> is resistant to spectinomycin and carbenicillin.
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NEED LINK
 
 
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<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section2">Results </a> (May need to open in a new tab.)
 
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*Determined that the microbe <i>Ga. hansenii</i> is essential for the fermentation of kombucha.
 
*Determined that the microbe <i>Ga. hansenii</i> is essential for the fermentation of kombucha.
 
*Determined that two distinct strains of the yeast <i>Lachancea fermentati</i> are necessary for the fermentation of kombucha, including one that appears to produce high quantities of C02.
 
*Determined that two distinct strains of the yeast <i>Lachancea fermentati</i> are necessary for the fermentation of kombucha, including one that appears to produce high quantities of C02.
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NEED LINK
 
 
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<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section3">Results </a> (May need to open in a new tab.)
 
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*Designed Golden Gate parts for the assembly of these genes into a functional construct.
 
*Designed Golden Gate parts for the assembly of these genes into a functional construct.
 
*Used a bromothymol blue assay to compare changes in pH resulting from fermentation in multiple strains of <i>Lachancea fermentati</i> isolated from our kombucha.
 
*Used a bromothymol blue assay to compare changes in pH resulting from fermentation in multiple strains of <i>Lachancea fermentati</i> isolated from our kombucha.
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NEED LINK
 
 
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<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section4">Results </a>( May need to open in a new tab.)
 
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<div id="section5"  class = "naviSection">
 
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<h2> Brazzein </h2>
 
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<P> One of the potential methods to create designer kombucha is to add a brazzein gene into the bacterial strains. Brazzein, a protein found in the pulp of the edible fruit of the African plant <i>Pentadiplandra brazzeana Baill</i>, is an extremely sweet substance<sup>1</sup>. It is 2,000 times sweeter than sucrose by weight. This makes it a healthy and economical alternative to sugar.  Commercial production of brazzein is limited, however, because it comes from a tropical plant. If it could be more easily harvested, it could be used to improve the flavor of various foods and drinks, including kombucha. By genetically engineering the brazzein gene into the bacteria in kombucha, the drink could be sweetened without adding sugar or excessive calories.  While still being a GMO product, this beverage would be low in sugar and could appeal to a health-conscious consumer.</p>
 
  
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<h3>References</h3>
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<div id="section6"  class = "naviSection">
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<ol type="1">
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<li>Yan, Sen et al. “Expression of Plant Sweet Protein Brazzein in the Milk of Transgenic Mice.” Ed. Xiao-Jiang Li. PLoS ONE 8.10 (2013): e76769. </li>
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<li>Brazzein protein structure acquired from European Bioinformatics Institute </li>
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</ol>
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<h2>pH Sensors</h2>
 
<h2>pH Sensors</h2>
  
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*Further characterized the P-atp2 Biobrick.
 
*Further characterized the P-atp2 Biobrick.
 
*Found literature describing three putative promoters in <i>Gluconobacter oxydans</i> that increase transcription under acidic conditions, and currently characterizing these sequences.
 
*Found literature describing three putative promoters in <i>Gluconobacter oxydans</i> that increase transcription under acidic conditions, and currently characterizing these sequences.
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NEED LINK
 
 
<html>
 
<html>
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<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section6">Results</a> (May need to open in a new tab.)
 
</div>
 
</div>
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<div id="section7"  class = "naviSection">
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<h2>Gellan Gum</h2>
 +
 +
</html>
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* Successfully made Gellan Gum plates from <i>Sphingomonas paucimobilis</i>
 +
* Successfully grew other bacteria on the Gellan Gum plates
 +
* Shared this DIY technology with the Texas Tech iGEM team
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<html>
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<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section7">Results </a> (May need to open in a new tab.)
 
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<p>Tell us about your project, describe what moves you and why this is something important for your team.</p>
 +
 
 +
 
 +
<h5>What should this page contain?</h5>
 +
<ul>
 +
<li> A clear and concise description of your project.</li>
 +
<li>A detailed explanation of why your team chose to work on this particular project.</li>
 +
<li>References and sources to document your research.</li>
 +
<li>Use illustrations and other visual resources to explain your project.</li>
 +
</ul>
  
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<div class="column full_size_inner">
 
  
 
</div>
 
</div>
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<div class="column full_size" >
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<h5>Advice on writing your Project Description</h5>
 +
 +
<p>
 +
We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.
 +
</p>
 +
 +
<p>
 +
Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year.
 +
</p>
 +
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</div>
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<div class="column half_size" >
 +
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<h5>References</h5>
 +
<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you thought about your project and what works inspired you.</p>
 +
 +
</div>
 +
 +
 +
<div class="column half_size" >
 +
<h5>Inspiration</h5>
 +
<p>See how other teams have described and presented their projects: </p>
 +
 +
<ul>
 +
<li><a href="https://2014.igem.org/Team:Imperial/Project"> Imperial</a></li>
 +
<li><a href="https://2014.igem.org/Team:UC_Davis/Project_Overview"> UC Davis</a></li>
 +
<li><a href="https://2014.igem.org/Team:SYSU-Software/Overview">SYSU Software</a></li>
 +
</ul>
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</div>
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  <h2>Demonstrate</h2>
 +
  <br>
 +
    <p> Click on one of the images below to learn more about our results! </p>
 +
    </div>
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<input type="image" src ="https://static.igem.org/mediawiki/2016/e/e7/T--Austin_UTexas--pHNavi.png" style="width:100%;" onclick="showOne('section6')" /><p>pH</p>
 
<input type="image" src ="https://static.igem.org/mediawiki/2016/e/e7/T--Austin_UTexas--pHNavi.png" style="width:100%;" onclick="showOne('section6')" /><p>pH</p>
 
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<h2>Kombucha Strains</h2>  
 
<h2>Kombucha Strains</h2>  
  
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*Identified strains of bacteria and yeast using rRNA gene sequencing.
 
*Identified strains of bacteria and yeast using rRNA gene sequencing.
 
*Characterized each of the isolated microbes to facilitate further experimentation.  
 
*Characterized each of the isolated microbes to facilitate further experimentation.  
−
NEED LINK
 
  
−
__NOTOC__
 
 
<html>
 
<html>
 +
<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section1">Results </a>
 
</div>
 
</div>
  
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<h2>Recapitulation</h2>
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<h2>Conjugation</h2>  
  
 
</html>
 
</html>
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*In a process called "recapitulation," we successfully created a kombucha-like culture by adding individual strains of microbes instead of a living culture containing the entire kombucha microbiome.
+
*Attempted conjugation with <i>G. oxydans</i>.
−
*Determined that the microbe <i>Ga. hansenii</i> is essential for the fermentation of kombucha.
+
*Performed minimum inhibitory concentration experiments between <i>G. oxydans</i> and spectinomycin, carbenicillin and kanamycin.
−
*Determined that two distinct strains of the yeast <i>Lachancea fermentati</i> are necessary for the fermentation of kombucha, including one that appears to produce high quantities of C02.
+
*Determined that <i>G. oxydans</i> is resistant to spectinomycin and carbenicillin.
−
NEED LINK
+
 
<html>
 
<html>
 +
<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section2">Results </a>
 
</div>
 
</div>
  
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<h2>Recapitulation</h2>
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<h2>Conjugation</h2>  
+
  
 
</html>
 
</html>
−
*Attempted conjugation with <i>G. oxydans</i>.
+
*In a process called "recapitulation," we successfully created a kombucha-like culture by adding individual strains of microbes instead of a living culture containing the entire kombucha microbiome.
−
*Performed minimum inhibitory concentration experiments between <i>G. oxydans</i> and spectinomycin, carbenicillin and kanamycin.
+
*Determined that the microbe <i>Ga. hansenii</i> is essential for the fermentation of kombucha.
−
*Determined that <i>G. oxydans</i> is resistant to spectinomycin and carbenicillin.
+
*Determined that two distinct strains of the yeast <i>Lachancea fermentati</i> are necessary for the fermentation of kombucha, including one that appears to produce high quantities of C02.
−
NEED LINK
+
 
<html>
 
<html>
 +
<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section3">Results </a>
 
</div>
 
</div>
  
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<h2>Ethanol</h2>
 
<h2>Ethanol</h2>
  
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*Designed Golden Gate parts for the assembly of these genes into a functional construct.
 
*Designed Golden Gate parts for the assembly of these genes into a functional construct.
 
*Used a bromothymol blue assay to compare changes in pH resulting from fermentation in multiple strains of <i>Lachancea fermentati</i> isolated from our kombucha.
 
*Used a bromothymol blue assay to compare changes in pH resulting from fermentation in multiple strains of <i>Lachancea fermentati</i> isolated from our kombucha.
−
NEED LINK
 
 
<html>
 
<html>
 +
<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section4">Results </a>
 
</div>
 
</div>
  
−
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<h2>pH Sensors</h2>
 
<h2>pH Sensors</h2>
  
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*Further characterized the P-atp2 Biobrick.
 
*Further characterized the P-atp2 Biobrick.
 
*Found literature describing three putative promoters in <i>Gluconobacter oxydans</i> that increase transcription under acidic conditions, and currently characterizing these sequences.
 
*Found literature describing three putative promoters in <i>Gluconobacter oxydans</i> that increase transcription under acidic conditions, and currently characterizing these sequences.
−
NEED LINK
 
 
<html>
 
<html>
−
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<a href = "https://2016.igem.org/Team:Austin_UTexas/Results#section6">Results </a></div>
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Latest revision as of 00:48, 20 October 2016

Demonstrate

For the gold medal “Demonstrate your work” requirement, we have shown that we can recreate kombucha from scratch by adding isolated strains of microbes to tea media. Our tea media, made with water, black tea, and sucrose, simulates the starting mixture used to brew kombucha. We have performed extensive recapitulation trials to determine which strains of yeast and bacteria are necessary for brewing kombucha. Cataloguing these vital strains is a necessary step toward modifying the starting population to create a “designer beverage” with a variety of properties that could benefit kombucha consumers and manufacturers outside the lab.

Click the images below to learn more about our results!

Kombucha Strains

Conjugation

Recapitulation

Ethanol

pH

Gellan Gum