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Unfortunately, at the time of the upload of this page there are no actual working results based off of the BAITSwitch. However, we are currently waiting to see the results of a Western Blot which may lead to the Bioorthoginal Auxin Inducible Trigger Switch degrading GFP in a reasonable timeframe.
Template:Nevada/ResultsandFutureApplication
From 2014.igem.org
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| - | <p>As seen above, we predict that when auxin is added to a yeast strain that contains Tir1 and IAA-tagged GFP, we will see the amount of GFP to rapidly decrease. With this in mind, it is possible that we will not see the amount of GFP decrease in the system that contains Coi1 and the IAAtagged GFP. | + | <p>As seen above, we predict that when auxin is added to a yeast strain that contains Tir1 and IAA-tagged GFP, we will see the amount of GFP to rapidly decrease. With this in mind, it is possible that we will not see the amount of GFP decrease in the system that contains Coi1 and the IAAtagged GFP.</br> |
<img src="https://static.igem.org/mediawiki/2014/c/c3/Team_nevada_igem_2014_amount_of_gfp_with_addition_of_auxin.png"> | <img src="https://static.igem.org/mediawiki/2014/c/c3/Team_nevada_igem_2014_amount_of_gfp_with_addition_of_auxin.png"> | ||
Furthermore, predict that with varying amounts of Auxin added to the appropriate system, we will also see a difference in the amount of GFP degraded within time. As we add higher amounts of Auxin to a yeast strain that contains Tir1 and IAA-tagged GFP, we will see the amount of GFP decrease even more rapidly. | Furthermore, predict that with varying amounts of Auxin added to the appropriate system, we will also see a difference in the amount of GFP degraded within time. As we add higher amounts of Auxin to a yeast strain that contains Tir1 and IAA-tagged GFP, we will see the amount of GFP decrease even more rapidly. | ||
Revision as of 23:30, 17 October 2014
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The BAITswitch
The Bioorthoginal Auxin Induceable Trigger Switch
As seen above, we predict that when auxin is added to a yeast strain that contains Tir1 and IAA-tagged GFP, we will see the amount of GFP to rapidly decrease. With this in mind, it is possible that we will not see the amount of GFP decrease in the system that contains Coi1 and the IAAtagged GFP.
Furthermore, predict that with varying amounts of Auxin added to the appropriate system, we will also see a difference in the amount of GFP degraded within time. As we add higher amounts of Auxin to a yeast strain that contains Tir1 and IAA-tagged GFP, we will see the amount of GFP decrease even more rapidly.
Furthermore, the system can work in parallel with the Coronatine system. Here when Coronatine is added to a system containing Tir1 and the IAA-tagged GFP, we should see the same effects of as the Auxin system.
Jaz1 is a degron tag utilized in the coronatine pathway. The protein of interest is tagged with Jaz1; when coronatine is added to the system, the protein and its tag are linked to the E3 ubiquitin ligase, COI1. Click the picture or the name below to be taken to the parts registry page.
RFP-Jaz1 is a degron tag utilized in the coronatine pathway. The protein of interest is tagged with RFP-Jaz1; when coronatine is added to the system, the protein and its tag are linked to the E3 ubiquitin ligase, COI1. Click the picture or the name below to be taken to the parts registry page.
Jaz6 is a degron tag utilized in the coronatine pathway. The protein of interest is tagged with Jaz6; when coronatine is added to the system, the protein and its tag are linked to the E3 ubiquitin ligase, COI1. Click the picture or the name below to be taken to the parts registry page.
COI-1 or Coronatine Insensitive-1 recruits JAZ-1 or JAZ-6 tagged substrates in the presence of Coronatine for degradation by the Proteasome.
TiRI is an ubuquitinase E3 that recognizes AID-tagged proteins and trigger their ubiquitination for further degradation. This part works with K812011 which is a AID tagged GFP. OsTirI is a protein comming from rice that have been identified as working in mammalian cells and yeast. This part takes place in a device pattented by Kanemaki Masato, Kakimoto Tatsuo, Nishimura Kohei, Takisawa Haruhiko and Fukagawa Tatsuo for yeast and mammalian cells use . However the patent does not cover the use for oviparian such as frogs and chicken.
This part is a GFP coding sequence fused to an AID tag for its recognition by the ubiquitinase E3 that induces its degradation in the presence of auxin in the eukaryote cell. This is designed to be used in pSB1C3 vector. This part is a part of a device patented by Kanemaki Masato, Kakimoto Tatsuo, Nishimura Kohei, Takisawa Haruhiko and Fukagawa Tatsuo for yeast and mammalian cells use.
