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

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[[File:T--Austin_UTexas--Patp2Results.png|thumb|right|500px| Figure 4. Spun down P-atp2 constructs compared to controls in pH6-9. There is no clear gradient change in color expression. Credit: Ian Overman and Alex Alario]]
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[[File:T--Austin_UTexas--Patp2Results.png|thumb|right|600px| Figure 4. Spun down P-atp2 constructs compared to controls in pH6-9. There is no clear gradient change in color expression. Credit: Ian Overman and Alex Alario]]
 
[[File:T--Austin_UTexas--BCPtube.png|thumb|left|150px| Figure 5. amilCP expressed in <i> E. coli </i> and in liquid LB. Credit: Riya Sreenivasan]]
 
[[File:T--Austin_UTexas--BCPtube.png|thumb|left|150px| Figure 5. amilCP expressed in <i> E. coli </i> and in liquid LB. Credit: Riya Sreenivasan]]
  
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<p>The P-atp2 promoter, native to the bacterium <i>Corynebacterium glutamicum</i> is reportedly induced at pH 7, to pH 9 (<a href="https://2015.igem.org/Team:BIT-China/Parts">BIT-China-2015</a> and <a href="http://parts.igem.org/Part:BBa_K1675021">BBa_K1675021</a>).<sup>1</sup> Utilizing the blue chromoprotein (<a href="http://partsregistry.org/Part:BBa_K592009">BBa_K592009</a>), a test was designed in which a plasmid containing the P-atp2 promoter with the blue chromoprotein was grown alongside an <i>E. coli</i> line that contained a plasmid with just the blue chromoprotein. We expected to see constant blue chromoprotein production in the control series (those that lacked P-atp2) and a visual increase in blue chromoprotein as the pH was raised from 6 to 9 in the cells that contained the P-atp2 construct. The construct utilized as a control can be found on the iGEM registry <a href="http://parts.igem.org/Part:BBa_2097001">BBa_K2097001</a> as as in figure 5.</p>
 
<p>The P-atp2 promoter, native to the bacterium <i>Corynebacterium glutamicum</i> is reportedly induced at pH 7, to pH 9 (<a href="https://2015.igem.org/Team:BIT-China/Parts">BIT-China-2015</a> and <a href="http://parts.igem.org/Part:BBa_K1675021">BBa_K1675021</a>).<sup>1</sup> Utilizing the blue chromoprotein (<a href="http://partsregistry.org/Part:BBa_K592009">BBa_K592009</a>), a test was designed in which a plasmid containing the P-atp2 promoter with the blue chromoprotein was grown alongside an <i>E. coli</i> line that contained a plasmid with just the blue chromoprotein. We expected to see constant blue chromoprotein production in the control series (those that lacked P-atp2) and a visual increase in blue chromoprotein as the pH was raised from 6 to 9 in the cells that contained the P-atp2 construct. The construct utilized as a control can be found on the iGEM registry <a href="http://parts.igem.org/Part:BBa_2097001">BBa_K2097001</a> as as in figure 5.</p>
 
<p>However, as seen in figure 4, no clear change in color expression appears in the experimental trials, suggesting a lack of sensitivity of the P-atp2 promoter.</p>
 
<p>However, as seen in figure 4, no clear change in color expression appears in the experimental trials, suggesting a lack of sensitivity of the P-atp2 promoter.</p>
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<h4>GOX Sequences as Putative Promoters</h4>
 
<h4>GOX Sequences as Putative Promoters</h4>
 
<p>Three endogenous upstream regions of loci on the <i>Gluconobacter oxydans</i>  chromosome were reported to show increased mRNA synthesis as pH decreased, were isolated and obtained, as seen in table 1.<sup>2</sup> Using Golden Gate assembly, these putative promoters have been placed on the Golden Gate entry vector pYTK001 for later use. By utilizing these pH-sensitive promoters with different reporters and transforming them into multiple organisms in kombucha, the visualization of the microbes and their location in kombucha would be possible.<sup>4</sup> This will serve as a stepping stone into further understanding how the microbiome of kombucha changes as it brews as well as determining organism concentration specific times during the brewing process.</p>
 
<p>Three endogenous upstream regions of loci on the <i>Gluconobacter oxydans</i>  chromosome were reported to show increased mRNA synthesis as pH decreased, were isolated and obtained, as seen in table 1.<sup>2</sup> Using Golden Gate assembly, these putative promoters have been placed on the Golden Gate entry vector pYTK001 for later use. By utilizing these pH-sensitive promoters with different reporters and transforming them into multiple organisms in kombucha, the visualization of the microbes and their location in kombucha would be possible.<sup>4</sup> This will serve as a stepping stone into further understanding how the microbiome of kombucha changes as it brews as well as determining organism concentration specific times during the brewing process.</p>

Revision as of 22:54, 18 October 2016

Austin_UTexas

Results


Click on one of the images below to learn more about our results!




Kombucha Strains

Conjugation

Recapitulation

Ethanol

pH Sensors