Team:Washington/Parts

From 2011.igem.org

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| '''Make It:  Diesel Production''' <br><br> We designed and constructed a modular alkane production BioBrick, the PetroBrick, to generate alkanes, the main constituent of diesel.  By expressing this BioBrick in ''E. coli'', we were able to produce alkanes of yields over 100 mg/mL.
| '''Make It:  Diesel Production''' <br><br> We designed and constructed a modular alkane production BioBrick, the PetroBrick, to generate alkanes, the main constituent of diesel.  By expressing this BioBrick in ''E. coli'', we were able to produce alkanes of yields over 100 mg/mL.
| '''Break It:  Gluten Destruction''' <br><br> Gluten intolerance stems from an inappropriate immune response to PQLP, the most common motif in the immunogenic peptide.  We reengineered a protease, active at low pH, for strongly enhanced activity against PQLP.
| '''Break It:  Gluten Destruction''' <br><br> Gluten intolerance stems from an inappropriate immune response to PQLP, the most common motif in the immunogenic peptide.  We reengineered a protease, active at low pH, for strongly enhanced activity against PQLP.
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| '''iGEM Toolkits: Gibson Assembly and Magnetosome''' <br><br> We built two iGEM toolkits. In the first we created five Gibson Cloning versions of traditional biobrick vectors, and in the second we cloned the genes essential for magnetosome formation and transformed two into ''E. coli''
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| '''iGEM Toolkits: Gibson Assembly and Magnetosomes''' <br><br> We built two iGEM toolkits. In the first we created five Gibson Cloning versions of traditional biobrick vectors, and in the second we cloned the genes essential for magnetosome formation and transformed two into ''E. coli''
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Revision as of 01:32, 29 September 2011


Data Page


An overview of the 2011 UW iGEM team's summer projects

Washington Spacer.jpg UW Diesel Front Page.png Washington Spacer.jpg UW Toolkits Front Page.png Washington Spacer.jpg UW Gluten Front Page.png Washington Spacer.jpg

Make It: Diesel Production

We designed and constructed a modular alkane production BioBrick, the PetroBrick, to generate alkanes, the main constituent of diesel. By expressing this BioBrick in E. coli, we were able to produce alkanes of yields over 100 mg/mL.
Break It: Gluten Destruction

Gluten intolerance stems from an inappropriate immune response to PQLP, the most common motif in the immunogenic peptide. We reengineered a protease, active at low pH, for strongly enhanced activity against PQLP.
iGEM Toolkits: Gibson Assembly and Magnetosomes

We built two iGEM toolkits. In the first we created five Gibson Cloning versions of traditional biobrick vectors, and in the second we cloned the genes essential for magnetosome formation and transformed two into E. coli


Data Summary

Data for Favorite New Parts

Diesel Production

1, 2. BBa_K590025: The PetroBrick - A modular and open platform for the biological production of diesel fuel. The PetroBrick consists of AAR and ADC, each behind a standard Elowitz RBS. All of this is under regulation by a high constitutive promoter in pSB1C3.

Gluten Destruction

3. BBa_K590087: KumaMax- A modified version of the enzyme Kumamolisin, a protease ofthe sedolisin family native to Alicyclobacillus sendaiensis known to be active at low pH and elevated temperatures. To Kumamolisin, the mutations N291D, G319S D358G, D368H increase activity to the PQLP peptide, an antigenic epitope in gliadin, 118-fold.

Gibson Assembly Toolkit

4. BBa_K590010: pGA1A3, BBa_K590011: pGA1C3, BBa_K590012: pGA4C5, BBa_K590013: pGA4A5, BBa_K590014: pGA3K3 - These are plasmid backbones based on the bglBrick standard (BBF RFC 21) and optimized for use in Gibson assembly. These vectors are suitable replacements for the equivalent pSB vectors for iGEM teams using Gibson cloning to assemble their constructs.

Magnetosome Toolkit

5. BBa_K590015: sfGFP_mamK_pGA1C3 - This part consists of the mamK gene from Magnetospirillum magneticum strain AMB-1, as a superfolder GFP fusion, in the pGA1C3 backbone. MamK creates an actin-like filament that orients itself along the long-axis of the cell and acts as the scaffold for the alignment of magnetosome vesicles.
6. BBa_K590016 sfGFP_mamI_pGA1C3 - This part consists of mamI gene from Magnetospirillum magneticum strain AMB-1, as a superfolder GFP fusion, in the pGA1C3 backbone. MamI is a membrane-localized protein required for magnetosome vesicle formation that also binds the poly-MamK filament.

Data for Existing Parts

7. K314100: High Constitutive Expression Cassette (Washington, iGEM 2010) - We used this part to express our Petrobrick, found that it works well for expression, and entered this information in the part experience page.
8. pSB1A3 - As part of the Gibson Vector Toolkit we characterized the cloning efficiency of this plasmid backbone for Gibson assembly, using the prefix and suffix regions as primers. We found that pSB1A3 had a proper insert efficiency of 11%, compared to 99% for the equivalent Gibson-optimized pGA1A3 vector.

Improved Parts

9. BBa_K590059, BBa_K590060, BBa_K590061: LuxC, D, and E (Cambridge, iGEM 2010) - Formerly part of the LuxBrick the genes luxC, D, and E were not separated or codon-optimized. We codon-optimized these genes and put them under the control of standard Elowitz RBS's (B0034). This was accomplished by the Alternate Aldehyde branch of the Alkane Production team.

All Submitted Parts

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