Difference between revisions of "Team:WashU StLouis/Proof"

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<img src = "https://static.igem.org/mediawiki/2016/b/be/T--WashU_StLouis--BiotinYalegraph2.png" style="width : 700px">
 
<img src = "https://static.igem.org/mediawiki/2016/b/be/T--WashU_StLouis--BiotinYalegraph2.png" style="width : 700px">
 
<img src = "https://static.igem.org/mediawiki/2016/c/c7/T--WashU_StLouis--BiotinDH10Bgraph.png" style="width : 700px">
 
<img src = "https://static.igem.org/mediawiki/2016/c/c7/T--WashU_StLouis--BiotinDH10Bgraph.png" style="width : 700px">
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iGEM teams are great at making things work! We value teams not only doing an incredible job with theoretical models and experiments, but also in taking the first steps to make their project real.  
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After identifying the genes and constructing the electron donor overproducing  plasmids, a proper assay was necessary in order to see if our new cells produced usable forms of both electron donors. Not only was it shown that the products were overproduced, but also significant data supported the claim that these products were usable in protein synthesis.  
 
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<h4> What should we do for our proof of concept? </h4>
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After analyzing the results of our absorbance data of electron donors, it was decided to go a step further to ensure that these construct produced usable electron donors. A biotin assay was found that offered an indirect way of measuring the amount of reduced electron donors present. Biotin, commonly know as vitamin B7, is produced by a pathway that uses reduced flavodoxin/ferredoxin,specifically the SAM pathway that turns dethiobiotin to biotin (see Figure 1).
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You can assemble a device from BioBricks and show it works. You could build some equipment if you're competing for the hardware award. You can create a working model of your software for the software award. Please note that this not an exhaustive list of activities you can do to fulfill the gold medal criterion. As always, your aim is to impress the judges!
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[Biotin Pathway]
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By measuring the amount of biotin in the new cells, and recognizing the 2:1 stoichiometric relationship between reduced electron donors and biotin molecules, the amount of electron donors present and used can be indirectly measured. Results showed that __________, as seen in Figure __ (this will be the graph we have).
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[graph 1]
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[graph 2]
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Figure 1: Biotin synthesis pathway in E Coli
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http://pubs.rsc.org/en/content/articlelanding/2011/mb/c1mb05022b#!divAbstract
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https://tools.thermofisher.com/content/sfs/gallery/high/16174-Lucif-Firefly-rxn.jpg
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Revision as of 17:20, 17 October 2016

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Proof of Concept

Showing that our stuff could make things

After identifying the genes and constructing the electron donor overproducing plasmids, a proper assay was necessary in order to see if our new cells produced usable forms of both electron donors. Not only was it shown that the products were overproduced, but also significant data supported the claim that these products were usable in protein synthesis.

After analyzing the results of our absorbance data of electron donors, it was decided to go a step further to ensure that these construct produced usable electron donors. A biotin assay was found that offered an indirect way of measuring the amount of reduced electron donors present. Biotin, commonly know as vitamin B7, is produced by a pathway that uses reduced flavodoxin/ferredoxin,specifically the SAM pathway that turns dethiobiotin to biotin (see Figure 1).

[Biotin Pathway]

By measuring the amount of biotin in the new cells, and recognizing the 2:1 stoichiometric relationship between reduced electron donors and biotin molecules, the amount of electron donors present and used can be indirectly measured. Results showed that __________, as seen in Figure __ (this will be the graph we have).

[graph 1]
[graph 2] Figure 1: Biotin synthesis pathway in E Coli http://pubs.rsc.org/en/content/articlelanding/2011/mb/c1mb05022b#!divAbstract https://tools.thermofisher.com/content/sfs/gallery/high/16174-Lucif-Firefly-rxn.jpg