Difference between revisions of "Team:Freiburg/Proof"
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| − | <h5 | + | <h5 style="text-align: center"> Proof of concept </h5> |
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| − | + | <div class='imgDiv'> | |
| − | + | <img id="c1Img" src="https://static.igem.org/mediawiki/2016/d/d1/T--Freiburg--Proof_of_Concept_Front1.png"> | |
| + | </div> | ||
| + | <div class='txtDiv'> | ||
| + | <div>We produced a carrier that is suitable for the display of proteins on its surface. | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="Arrow"> | ||
| + | <img id="c1Arrow" src="https://static.igem.org/mediawiki/2016/b/bd/T--Freiburg--gtRight.png"> | ||
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| − | + | <img src="https://static.igem.org/mediawiki/2016/2/2e/T--Freiburg--Proof_of_Concept_Figure1.png"> | |
| − | + | </div> | |
| − | + | <div class='txt2Div'> | |
| − | + | <div>We were able to produce engineered spores from B. subtilis and establish protocols for its purification. | |
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| − | + | <div class='resultDiv'> | |
| − | + | <img src="https://static.igem.org/mediawiki/2016/b/b0/T--Freiburg--Proof_of_Concept_Figure2.png | |
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| − | + | <div>We verified the display of heterologous proteins containing an epitope tag on the surface of the spores by immunostaining and flow cytometry analysis. Read more about the production of Nanocillus. | |
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| − | + | <img id="c2Img" src="https://static.igem.org/mediawiki/2016/c/c0/T--Freiburg--Proof_of_Concept_Front2.png"> | |
| − | + | </div> | |
| − | + | <div class='txtDiv'> | |
| − | + | <div>We achieved the display of nanobodies as binding moiety and showed their functionality towards a target. | |
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| + | <img id="c2Arrow" src="https://static.igem.org/mediawiki/2016/b/bd/T--Freiburg--gtRight.png"> | ||
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| − | + | <img src="https://static.igem.org/mediawiki/2016/4/4f/T--Freiburg--Proof_of_Concept_Figure3.png"> | |
| − | + | </div> | |
| − | + | <div class='txt2Div'> | |
| − | + | <div>The anti-GFP nanobody was fusion on spore coat proteins and displayed on their surface. We verified the binding of the target GFP to the displayed nanobody by flow cytometry analysis. Learn more about targeting. | |
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| − | + | </div> | |
| − | + | <div class='txtDiv'> | |
| − | + | <div>We accomplished the display of functional glutathione S-transferase on the spores, an enzyme playing a crucial role in the activation of prodrugs. | |
| + | </div> | ||
| + | </div> | ||
| + | <div class="Arrow"> | ||
| + | <img id="c3Arrow" src="https://static.igem.org/mediawiki/2016/b/bd/T--Freiburg--gtRight.png"> | ||
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| − | + | <img src="https://static.igem.org/mediawiki/2016/4/4f/T--Freiburg--Proof_of_Concept_Figure4.png"> | |
| − | + | </div> | |
| − | + | <div class='txt2Div'> | |
| − | + | <div>Glutathione S-transferase converts the prodrug azathioprine to its active form 6-mercaptopurine. We displayed the enzyme on the spore surface and validated its functionality by a colorimetric GST assay. Read more about the drug delivery. | |
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Revision as of 21:57, 17 October 2016
Proof of concept
We produced a carrier that is suitable for the display of proteins on its surface.
We were able to produce engineered spores from B. subtilis and establish protocols for its purification.
We verified the display of heterologous proteins containing an epitope tag on the surface of the spores by immunostaining and flow cytometry analysis. Read more about the production of Nanocillus.
We achieved the display of nanobodies as binding moiety and showed their functionality towards a target.
The anti-GFP nanobody was fusion on spore coat proteins and displayed on their surface. We verified the binding of the target GFP to the displayed nanobody by flow cytometry analysis. Learn more about targeting.
We accomplished the display of functional glutathione S-transferase on the spores, an enzyme playing a crucial role in the activation of prodrugs.
Glutathione S-transferase converts the prodrug azathioprine to its active form 6-mercaptopurine. We displayed the enzyme on the spore surface and validated its functionality by a colorimetric GST assay. Read more about the drug delivery.
