Difference between revisions of "Team:Freiburg/Proof"
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<h5 style="text-align: center; margin-bottom:0px;"> Proof of Concept </h5> | <h5 style="text-align: center; margin-bottom:0px;"> Proof of Concept </h5> | ||
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| − | <div>We were able to produce engineered spores from B. subtilis and establish protocols for its purification. | + | <div>We were able to produce engineered spores from <i>B. subtilis</i> and establish protocols for its purification. |
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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 | + | <div>We verified the display of heterologous proteins containing an epitope tag on the surface of the spores by immunostaining and flow cytometry. Read more about the production of Nanocillus <a href="https://2016.igem.org/Team:Freiburg/Production_Nanocillus" target="_blank" style="text-align: center;font-size:20px;color:#e8a126">here</a>. |
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| − | <div>The anti-GFP nanobody was | + | <div>The anti-GFP nanobody was fused on spore coat proteins and displayed on their spore surface. Using statistical analysis we verified the binding of the target GFP to the displayed nanobody by flow cytometry. Learn more about our functional binding spore <a href="https://2016.igem.org/Team:Freiburg/Binding" target="_blank" style="text-align: center;font-size:20px;color:#e8a126">here</a>. |
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<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. | <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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| + | <br> Click <a href="https://2016.igem.org/Team:Freiburg/Delivery" target="_blank" style="text-align: center;font-size:20px;color:#e8a126">here</a> for more information. | ||
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Latest revision as of 01:55, 20 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. Read more about the production of Nanocillus here.
We achieved the display of nanobodies as binding moiety and showed their functionality towards a target.
The anti-GFP nanobody was fused on spore coat proteins and displayed on their spore surface. Using statistical analysis we verified the binding of the target GFP to the displayed nanobody by flow cytometry. Learn more about our functional binding spore here.
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.
Click here for more information.
Click here for more information.
