Difference between revisions of "Team:BostonU/Parts"

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<p><a href="http://parts.igem.org/Part:BBa_K1733001" style="color:blue;">ABI</a> and <a href="http://parts.igem.org/Part:BBa_K1733002" style="color:blue;">PYL</a></p>
 
<p><a href="http://parts.igem.org/Part:BBa_K1733001" style="color:blue;">ABI</a> and <a href="http://parts.igem.org/Part:BBa_K1733002" style="color:blue;">PYL</a></p>
  
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These parts contain the ABI (ABA insensitive 1) and PYL (pyrabactin resistance like) protein domains that will dimerize with each other in the presence of abscisic acid. By fusing these dimerization domains to inert halves of a protein, we were able to control protein function and induce dimerization, and protein activity, with the addition of ABA (abscisic acid). We were therefore able to regulate protein activity by taking advantage of these dimerization domains.
 
These parts contain the ABI (ABA insensitive 1) and PYL (pyrabactin resistance like) protein domains that will dimerize with each other in the presence of abscisic acid. By fusing these dimerization domains to inert halves of a protein, we were able to control protein function and induce dimerization, and protein activity, with the addition of ABA (abscisic acid). We were therefore able to regulate protein activity by taking advantage of these dimerization domains.
  
 
Both ABI and PYL are found in plants<sup>1</sup>. Thus, it can be tested in mammalian cells, as the ABI-PYL-ABA system is completely orthogonal. We tested this system with our split integrases in mammalian cells.
 
Both ABI and PYL are found in plants<sup>1</sup>. Thus, it can be tested in mammalian cells, as the ABI-PYL-ABA system is completely orthogonal. We tested this system with our split integrases in mammalian cells.
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We used this domain in our experiment to construct split protein complexes. We split the TP901-1 protein and fused the halves to ABI and PYL. We then added abscisic acid into the system to induce dimerization of the domains and the protein halves, and measured the protein activity TP901-1 afterwards. We were able to characterize some split sites as functional, as we regained the TP901-1 activity after inducing dimerization. One functional split site is shown below:
 
We used this domain in our experiment to construct split protein complexes. We split the TP901-1 protein and fused the halves to ABI and PYL. We then added abscisic acid into the system to induce dimerization of the domains and the protein halves, and measured the protein activity TP901-1 afterwards. We were able to characterize some split sites as functional, as we regained the TP901-1 activity after inducing dimerization. One functional split site is shown below:
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<a href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2015&group=BostonU" style="color:blue;">Our parts page</a>
 
<a href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2015&group=BostonU" style="color:blue;">Our parts page</a>
 
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<h4 style="font-size:16px; text-align:center;">Citations</h4>
 
<h4 style="font-size:16px; text-align:center;">Citations</h4>
 
<ol style="font-size:12px;">
 
<ol style="font-size:12px;">
 
<li>Liang, Fu-Sen, Ho, Wen Qi, Crabtree, Gerald R., “Engineering the ABA Stress Pathway for Regulation of Induced Proximity”, Sci Signal, 2011.</li>
 
<li>Liang, Fu-Sen, Ho, Wen Qi, Crabtree, Gerald R., “Engineering the ABA Stress Pathway for Regulation of Induced Proximity”, Sci Signal, 2011.</li>
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Revision as of 17:09, 18 September 2015

Parts

Parts

We submitted the following three parts of our experiment to the iGEM registry:

Dimerization Domains:

ABI and PYL

These parts contain the ABI (ABA insensitive 1) and PYL (pyrabactin resistance like) protein domains that will dimerize with each other in the presence of abscisic acid. By fusing these dimerization domains to inert halves of a protein, we were able to control protein function and induce dimerization, and protein activity, with the addition of ABA (abscisic acid). We were therefore able to regulate protein activity by taking advantage of these dimerization domains. Both ABI and PYL are found in plants1. Thus, it can be tested in mammalian cells, as the ABI-PYL-ABA system is completely orthogonal. We tested this system with our split integrases in mammalian cells.

We used this domain in our experiment to construct split protein complexes. We split the TP901-1 protein and fused the halves to ABI and PYL. We then added abscisic acid into the system to induce dimerization of the domains and the protein halves, and measured the protein activity TP901-1 afterwards. We were able to characterize some split sites as functional, as we regained the TP901-1 activity after inducing dimerization. One functional split site is shown below:

Recombination Directionality Factors:

orf7

Here is a link to our parts: Our parts page

Citations

  1. Liang, Fu-Sen, Ho, Wen Qi, Crabtree, Gerald R., “Engineering the ABA Stress Pathway for Regulation of Induced Proximity”, Sci Signal, 2011.