Difference between revisions of "Team:UAlberta/Improve"

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<h1>Improve a Part / Project</h1>
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<h2>Improved BioBrick Part: BBa_K2779912</h2>
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<p>Team UAlberta’s cloning work relied heavily on the use of fluorescent markers to assemble our desired constructs, either with fluorescence serving as the background or by incorporating fluorescence into our desired constructs. Additionally, as part of our characterization of our engineered pathway, we wanted to be able to express each enzyme and purify it for in vitro characterization and experimentation.</p>
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<p>To accomplish this, Team UAlberta decided to use an araBAD-based expression system because:</p>
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<ul>
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<li>It is inducible by L-arabinose and is tightly regulated by araC</li>
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<li>It consists of a strong promoter  for increased expression</li>
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<li>It is compatible with our preferred workhorse strain, DH10B, allowing us to perform our cloning and expression work in the same strain</li>
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<li>It is more cost-effective since L-arabinose is more economical than IPTG.</li>
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</ul>
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<p>When we initially looked through the repository, we found that while BBa_K1602055, which had GFP under the control of araC/pBAD, was the closest to our needs, there was no easy way for us to replace their inserted GFP with our desired inserts, and there was no way for us to collect the expressed enzymes.</p>
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<figure class="figure">
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              <img src="https://static.igem.org/mediawiki/2018/6/6d/T--UAlberta--BBa_K1602055.svg
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" class="figure-img img-fluid rounded" alt="...">
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              <figcaption class="figure-caption text-right"><strong>Figure 1:</strong> A schematic of BBa_K1602055.</figcaption>
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Revision as of 01:26, 18 October 2018

...

Improve a Part / Project

Improved BioBrick Part: BBa_K2779912

Team UAlberta’s cloning work relied heavily on the use of fluorescent markers to assemble our desired constructs, either with fluorescence serving as the background or by incorporating fluorescence into our desired constructs. Additionally, as part of our characterization of our engineered pathway, we wanted to be able to express each enzyme and purify it for in vitro characterization and experimentation.

To accomplish this, Team UAlberta decided to use an araBAD-based expression system because:

  • It is inducible by L-arabinose and is tightly regulated by araC
  • It consists of a strong promoter for increased expression
  • It is compatible with our preferred workhorse strain, DH10B, allowing us to perform our cloning and expression work in the same strain
  • It is more cost-effective since L-arabinose is more economical than IPTG.

When we initially looked through the repository, we found that while BBa_K1602055, which had GFP under the control of araC/pBAD, was the closest to our needs, there was no easy way for us to replace their inserted GFP with our desired inserts, and there was no way for us to collect the expressed enzymes.

...
Figure 1: A schematic of BBa_K1602055.