Difference between revisions of "Team:BostonU/Attributions/Experimental Workflow"
| Line 20: | Line 20: | ||
<p>Below are the graphs generated by our MATLAB model for each protein that we aimed to split. The split sites are indicated in black.</p> | <p>Below are the graphs generated by our MATLAB model for each protein that we aimed to split. The split sites are indicated in black.</p> | ||
<br> | <br> | ||
| − | <p>Here are the two candidate integrase proteins. Learn more about our experimental motivation for developing dimerizable integrase proteins here.</p> | + | <p>Here are the two candidate integrase proteins. Learn more about our experimental motivation for developing dimerizable integrase proteins <a href="https://2015.igem.org/Team:BostonU/App_1/Motivation" style="color:#FF9966;">here</a>.</p> |
<br> | <br> | ||
<center><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/thumb/c/c4/Tp901_split_sites.png/800px-Tp901_split_sites.png.jpeg" /><nbp><nbp><nbp><nbp><nbp><nbp><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/thumb/5/56/PhiC31_split_sites.png/800px-PhiC31_split_sites.png.jpeg" /><center> | <center><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/thumb/c/c4/Tp901_split_sites.png/800px-Tp901_split_sites.png.jpeg" /><nbp><nbp><nbp><nbp><nbp><nbp><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/thumb/5/56/PhiC31_split_sites.png/800px-PhiC31_split_sites.png.jpeg" /><center> | ||
<br> | <br> | ||
| − | <p>Here are the two candidate RDF proteins. Learn more about our experimental motivation for developing dimerizable RDF proteins here.</p> | + | <p>Here are the two candidate RDF proteins. Learn more about our experimental motivation for developing dimerizable RDF proteins <a href="https://2015.igem.org/Team:BostonU/App_1/Motivation" style="color:#FF9966;">here</a>.</p> |
<br> | <br> | ||
<center><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/b/b1/Orf7jeffpng8.png" /><nbp><nbp><nbp><nbp><nbp><nbp><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/e/e6/Gp3jeffpng.png" /><center> | <center><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/b/b1/Orf7jeffpng8.png" /><nbp><nbp><nbp><nbp><nbp><nbp><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/e/e6/Gp3jeffpng.png" /><center> | ||
<br> | <br> | ||
| − | <p>Here is the saCas9 protein. Learn more about our experimental motivation for developing a dimerizable saCas9 protein here.</p> | + | <p>Here is the saCas9 protein. Learn more about our experimental motivation for developing a dimerizable saCas9 protein <a href="https://2015.igem.org/Team:BostonU/App_2/Motivation" style="color:#FF9966;">here</a>.</p> |
<br> | <br> | ||
<center><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/thumb/3/3f/Sacas9_split_sites.png/800px-Sacas9_split_sites.png.jpeg" /><center> | <center><img style="height:45%; width:45%;" src="https://static.igem.org/mediawiki/2015/thumb/3/3f/Sacas9_split_sites.png/800px-Sacas9_split_sites.png.jpeg" /><center> | ||
<br><br><br> | <br><br><br> | ||
</body> | </body> | ||
Revision as of 22:49, 18 September 2015
| Modeling | Dimerization Domains | Experimental Workflow |
After choosing potentially viable split sites and the dimerization domains, our team set out to clone a large set constructs and test them in HEK239T cells. For each indicated split site, we created 6 split pair variants - 3 using the different dimerization domains, and 3 using the same domains but in the opposite orientation. This means that for each split protein, we set out to clone 12 different plasmids. Ultimately, our construct set included over 400 constructs! We actually ended up cloning and testing around 200 of these.
Below are the graphs generated by our MATLAB model for each protein that we aimed to split. The split sites are indicated in black.
Here are the two candidate integrase proteins. Learn more about our experimental motivation for developing dimerizable integrase proteins here.
Here are the two candidate RDF proteins. Learn more about our experimental motivation for developing dimerizable RDF proteins here.
Here is the saCas9 protein. Learn more about our experimental motivation for developing a dimerizable saCas9 protein here.
