Difference between revisions of "Team:Bio Without Borders/Design"
| Line 6: | Line 6: | ||
<div class="image"><img src="https://static.igem.org/mediawiki/2018/5/55/T--Bio_Without_Borders--HorseshoeCrabs.png" alt="" /></div><div class="content"> | <div class="image"><img src="https://static.igem.org/mediawiki/2018/5/55/T--Bio_Without_Borders--HorseshoeCrabs.png" alt="" /></div><div class="content"> | ||
<h2>Creating Factor C<br /></h2> | <h2>Creating Factor C<br /></h2> | ||
| − | <p>We used an NEBuilder kit to combine two | + | <p>We used an NEBuilder kit to combine two sequences of the <i>Limulus polyphemus</i> that IDT sent us, as the whole sequence was too large to send as one part. We first confirmed that we could at the j04450 insert into the pSB1C3 backbone for submission. Once we confirmed this process, we then used the NEBuilder kit to put together the two G-Blocks of the Factor C sequence inside the pSCB1C3 backbone.</p> |
</div> | </div> | ||
</section> | </section> | ||
Revision as of 00:37, 18 October 2018
Creating Factor C
We used an NEBuilder kit to combine two sequences of the Limulus polyphemus that IDT sent us, as the whole sequence was too large to send as one part. We first confirmed that we could at the j04450 insert into the pSB1C3 backbone for submission. Once we confirmed this process, we then used the NEBuilder kit to put together the two G-Blocks of the Factor C sequence inside the pSCB1C3 backbone.
Creating a linker protein of the cellulose-binding domain (CBD) and GFP
This functions as the reporter protein which would demonstrate Factor C's ability to cleave.
Expressing Factor C through Pichia pastoris
After making our parts, we wanted to express the parts in an easy model organism like pichia.