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

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<h2> pH Sensors </h2>
 
<h2> pH Sensors </h2>
<P> Many of the microorganisms involved in the fermentation of kombucha produce acidic metabolites that lower the pH of the culture. Using pH-sensitive promoters to control the expression of reporter proteins, such as GFP or a chromoprotein, would allow visualization of the pH change. The promoters Cpx, P-atp2, and Cadc were selected as pH sensors to indicate pH in the neutral, basic, and acidic ranges, respectively [REFERENCES - 1 for each!!]. These constructs have been or will be transformed into <i>Escherichia coli</i> to confirm pH sensitivity prior to introduction to kombucha.</p><p> [THIS PARAGRAPH NEEDS SOME WORK--I have removed a lot, but I think a little fleshing out could help] Modification of <i>Gluconobacter oxydans</i>, a bacterium in kombucha, is also planned to avoid disturbing the kombucha microbiome. Three endogenous upstream regions of loci that were reported to show increased mRNA synthesis as pH decreased were obtained [REFERENCE PAPER!]. Golden Gate assembly is currently being used to quickly assemble these promoters upstream of Venus, [a BLANK FP, REFERENCE--pYTK paper!].<sup>1</sup> Once successful, these pH-sensitive promoters with different reporters will be used to visualize the different members of the kombucha microbiome overtime.</p>
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<p>Many of the microorganisms involved in the fermentation of kombucha produce acidic metabolites that lower the pH of the culture. Using pH-sensitive promoters to control the expression of reporter proteins, such as GFP or a chromoprotein, would allow visualization of the pH change. The promoters Cpx, P-atp2, and Cadc were selected as pH sensors to indicate pH in the neutral, basic, and acidic ranges, respectively.<sup>1,3,5,6</sup> These constructs have been or will be transformed into <i>Escherichia coli</i> to confirm pH sensitivity prior to introduction to kombucha and to see if these constructs could be utilized as sensors in mediums besides kombucha..</p>
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<p>Modification of <i>Gluconobacter oxydans</i>, a bacterium in kombucha, is also planned to avoid disturbing the kombucha microbiome. Three endogenous upstream regions of loci that were reported to show increased mRNA synthesis as pH decreased were obtained.<sup>2</sup> Golden Gate assembly is currently being used to quickly assemble these promoters upstream of Venus (pYTK033).<sup>4</sup> Once successful, these pH-sensitive promoters with different reporters will be used to visualize the different members of the kombucha microbiome overtime.</p>
  
 
<h3>Refrences</h3>
 
<h3>Refrences</h3>

Revision as of 22:25, 18 October 2016

Project Description


Gold Medal Part Characterization

The characterization of the BioBrick P-atp2 from the BIT-China-2015 team was done to see if P-atp2 could be utilized as a basic pH sensor. The results are found here and on the iGEM Registry page under experience, BBa_K1675021


Our Project

Kombucha is a beverage made when a symbiotic community of bacteria and yeast ferments sugared tea. Although kombucha has been consumed for thousands of years in the East, the drink has enjoyed a recent resurgence in popularity [REFERENCE-HOMEPAGE]. Several kombucha breweries operate in Austin, Texas, our team’s hometown. The role microbes play in the production of the beverage has led our team to wonder if synthetic biology could allow us to create “designer kombucha” with enhanced properties, such as more appealing flavors or additional nutrients. In order to do so, our team attempted to isolate the strains responsible for the fermentation of kombucha, identify them, genetically modify them, and add the individual strains into tea media to recreate the drink. We additionally considered potential applications of the ability to genetically modify the microbial population of kombucha, such as reducing the ethanol content of the beverage and improving taste with brazzein, a sweet-tasting protein. AS PART OF OUR HUMAN PRACTICES WE.... [SOMETHING HERE about GMO concerns and alternative direction(s)?]

Click the images below to learn more about our project!


Kombucha Strains

Conjugation

Recapitulation

Ethanol




Brazzein

pH Sensors