Difference between revisions of "Team:UofC Calgary/Judging"
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<li class="c-font-uppercase c-font-white">Part Characterization <a style="color: navy;" href="http://parts.igem.org/Part:BBa_K2008005"> BBa_K2008005</a></li> | <li class="c-font-uppercase c-font-white">Part Characterization <a style="color: navy;" href="http://parts.igem.org/Part:BBa_K2008005"> BBa_K2008005</a></li> | ||
<p class="c-font-white" style="font-size: 14px"> This part contains an active nonamer from the Bowman Birk Protease Inhibitor (BBI) with the addition of lysine, serine, cysteine, and isoleucine (KSCI) tag on the N-terminal end and a phenylalanine (F) tag on the C-terminal end to increase BBI solubility. A <i>B. subtilis</i> secretory signal peptide sequence is included to allow for secretion of the peptide directly into surrounding media. A transdermal tag <a style="color: navy;" href="http://parts.igem.org/Part:BBa_K1074000">(TD1, BBa_K1074000)</a> is fused to the N-terminus of BBI to allow for diffusion of the peptide across the skin. This coding sequence is under the control of the constitutive <i>B. subtilis </i>promoter pVeg <a style="color:navy;" href="http://parts.igem.org/Part:BBa_K143012">(BBa_K143012)</a> and a strong <i>B. subtilis</i> RBS <a style="color: navy;" href="http://parts.igem.org/Part:BBa_K780001">(BBa_K780001)</a>.</p> | <p class="c-font-white" style="font-size: 14px"> This part contains an active nonamer from the Bowman Birk Protease Inhibitor (BBI) with the addition of lysine, serine, cysteine, and isoleucine (KSCI) tag on the N-terminal end and a phenylalanine (F) tag on the C-terminal end to increase BBI solubility. A <i>B. subtilis</i> secretory signal peptide sequence is included to allow for secretion of the peptide directly into surrounding media. A transdermal tag <a style="color: navy;" href="http://parts.igem.org/Part:BBa_K1074000">(TD1, BBa_K1074000)</a> is fused to the N-terminus of BBI to allow for diffusion of the peptide across the skin. This coding sequence is under the control of the constitutive <i>B. subtilis </i>promoter pVeg <a style="color:navy;" href="http://parts.igem.org/Part:BBa_K143012">(BBa_K143012)</a> and a strong <i>B. subtilis</i> RBS <a style="color: navy;" href="http://parts.igem.org/Part:BBa_K780001">(BBa_K780001)</a>.</p> | ||
| − | <p class="c-font-white" style="font-size: 14px">We characterized this part by testing the effect of the BBI peptide in cell culturing experiments using both wildtype fibroblasts (1BR3) as well as colon cancer cells (HCT116). We further characterized the delivery capabilities of our TD1 tag by administering patch prototypes to mice, and then performing mass spectrometry on the mice blood a few days after patch administration to detect the presence of our peptide in the mice blood. You can find the results of these characterization experiments in the biotarget results section.</p> | + | <p class="c-font-white" style="font-size: 14px">We characterized this part by testing the effect of the BBI peptide in cell culturing experiments using both wildtype fibroblasts (1BR3) as well as colon cancer cells (HCT116). We further characterized the delivery capabilities of our TD1 tag by administering patch prototypes to mice, and then performing mass spectrometry on the mice blood a few days after patch administration to detect the presence of our peptide in the mice blood. You can find the results of these characterization experiments in the <a style="color: navy;" href="https://2016.igem.org/Team:UofC_Calgary/Results">biotarget results</a> section.</p> |
<br> | <br> | ||
<li class="c-font-uppercase c-font-white">Collaboration</li> | <li class="c-font-uppercase c-font-white">Collaboration</li> | ||
Revision as of 00:04, 20 October 2016
Bronze Requirements
- Register and Attend
- DELIVERABLES
- Wiki (Home Page)
- Poster
- Presentation
- Sample Submission
- Safety Form
- Judging Forms
- Parts Page
- Attributions
- New Part BBa_K2008003
We have registered for and will be attending the 2016 iGEM Jamboree in Boston!
You can find the contributions of each of our members as well as our numerous amazing mentors on our Attributions page
We designed and characterized a gene construct coding for the radioprotective peptide mBBI attached to a secretion tag, TD1 tag for secretion and transdermal delivery, as well as GFP for easy transformation recognition. Find it on our composite parts page here
Silver Requirements
- Part Characterization BBa_K2008005
- Collaboration
- Human Practices
This part contains an active nonamer from the Bowman Birk Protease Inhibitor (BBI) with the addition of lysine, serine, cysteine, and isoleucine (KSCI) tag on the N-terminal end and a phenylalanine (F) tag on the C-terminal end to increase BBI solubility. A B. subtilis secretory signal peptide sequence is included to allow for secretion of the peptide directly into surrounding media. A transdermal tag (TD1, BBa_K1074000) is fused to the N-terminus of BBI to allow for diffusion of the peptide across the skin. This coding sequence is under the control of the constitutive B. subtilis promoter pVeg (BBa_K143012) and a strong B. subtilis RBS (BBa_K780001).
We characterized this part by testing the effect of the BBI peptide in cell culturing experiments using both wildtype fibroblasts (1BR3) as well as colon cancer cells (HCT116). We further characterized the delivery capabilities of our TD1 tag by administering patch prototypes to mice, and then performing mass spectrometry on the mice blood a few days after patch administration to detect the presence of our peptide in the mice blood. You can find the results of these characterization experiments in the biotarget results section.
We have collaborated with various iGEM teams as well as certain local organizations for the improvement of our project and for the promotion of iGEM. See all of our collaborations on our Collaborations page.
Our project was continually informed by discussions with professionals in the field. See our human practices efforts on our Human Practices pages.
Gold Requirements
- Integrated Human Practices
- Improving upon a previous part BBa_K1444018
See how the feedback we got from professionals fed back into our project design on Human Practices Gold page.
B. subtilis is a gram-positive bacterial species which is widely used in molecular biology labs around the world. It has many benefits as a bacterium, but one of the major caveats is that it is very difficult transform under normal conditions. Transformation of B. subtilis will only occur under conditions of nutrient deprivation (energy starvation), which may not be the most ideal in the lab as it is very time-consuming, sensitive to precise timings, and can be unreliable.
In order to solve this problem, a previous University of Calgary iGEM team submitted the part ComK (which is a transcription factor that controls all of the genes for DNA uptake), to bypass the need for energy starvation in B. subtilis transformation (Zhang, 2010). This part contains a xylose-inducible promotor (pxylA), which means that once your bacterium is transformed with this construct, it will be readily transformable with other genes upon the addition of xylose to their nutrient broth.
Our team has modified this part by removing previously existing internal restriction sites, as well as adding flanking sequences of homology from the amyE locus to allow for chromosomal integration into the B. subtilis genome. This will drastically improve the transformation efficiency of the initial construct into eligible B. subtilis.
Previous team(s) that have worked with this part: University of Calgary iGEM 2014
