Difference between revisions of "Team:Washington/Parts"
Line 156: | Line 156: | ||
<html> | <html> | ||
We also have an automatically generated <a href="http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2015&group=Washington">Team Parts</a> page.</html> | We also have an automatically generated <a href="http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2015&group=Washington">Team Parts</a> page.</html> | ||
+ | |||
+ | <h2>Auxin</h2> | ||
+ | |||
+ | <p align = "justify"> This part, called AFB2, is an F-box protein that binds the common plant hormone auxin. F-box proteins contain an F-box domain that mediates degradation of other proteins by recognizing substrates for ubiquitination. </p> | ||
+ | |||
+ | <p align = "justify"> In our system, a dCas9-degron-repressor complex represses expression of lacZ. In the presence of auxin, the F-box protein and the presence of auxin, the AFB2 binds to auxin and auxin also binds to a degron; the F-box protein then recruits the ubiquitin ligase to degrade the dCas9-degron-repressor complex. When this occurs, lacZ is no longer repressed and production of beta galactosidase occurs. Beta galactosidase acts as a reporter for the presence of auxin. </p> | ||
<h2>Apatazyme</h2> | <h2>Apatazyme</h2> |
Revision as of 03:32, 19 September 2015
Biobricks
Parts in the registry
<groupparts>iGEM015 Washington</groupparts>
We also have an automatically generated Team Parts page.
Contents
Auxin
This part, called AFB2, is an F-box protein that binds the common plant hormone auxin. F-box proteins contain an F-box domain that mediates degradation of other proteins by recognizing substrates for ubiquitination.
In our system, a dCas9-degron-repressor complex represses expression of lacZ. In the presence of auxin, the F-box protein and the presence of auxin, the AFB2 binds to auxin and auxin also binds to a degron; the F-box protein then recruits the ubiquitin ligase to degrade the dCas9-degron-repressor complex. When this occurs, lacZ is no longer repressed and production of beta galactosidase occurs. Beta galactosidase acts as a reporter for the presence of auxin.
Apatazyme
yeVenus-TheoA; Coding sequence for enhanced YFP and theophylline-sensative aptazyme
The part contains the coding sequence for yeast enhanced yellow fluorescent protein (yeVenus), linked to it is a gene for a theophylline sensitive riboswitch aptazyme. The aptazyme portion of the transcript self-cleaves in the absence of theophylline and no YFP should be produced. The theophylline bound state stabilizes the transcript, which translates to the protein and fluorescence should be observed.
yeVenus-PEST-TheoA; Coding sequence for destabilized YFP Venus and theophylline-sensitive aptazyme
This part, yeVenus-PEST-TheoA, contains the coding sequence for a hybrid protein made of yeast enhanced yellow fluorescent protein (yeVenus) fused in frame with the CDS of the PEST-rich 178 c-terminal residues of Cln2, which targets the protein for ubiquitin dependent degradation. These coding sequences are fused with a gene for a theophylline sensitive riboswitch aptazyme. The aptazyme portion of the transcript self-cleaves in the absence of theophylline and no YFP should be produced. The theophylline bound state stabilizes the transcript, which translates to the protein and fluorescence should be observed.
</div> </div> </html>