From 2011.igem.org

Revision as of 17:11, 27 September 2011

An Overview of the 2011 UW iGEM Team's Summer Projects

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.

GibsonBricks and Magnetosome ToolKits We completed 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. [http://partsregistry.org/Part:BBa_K590025 BBa_K590025: The PetroBrick] - A modular and open platform for the biological production of diesel fuel. The PetroBrick consists of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590032 AAR] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590031 ADC], each behind a standard Elowitz RBS. All of this is under regulation by a high constitutive promoter in pSB1C3.

Gluten Destruction

3. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590087 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. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590010 BBa_K590010: pGA1A3_pLacGFP], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590011 BBa_K590011: pGA1C3_pLacGFP], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590012 BBa_K590012: pGA4C5_pLacGFP], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590013 BBa_K590013: pGA4A5_pLacGFP], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590014 BBa_K590014: pGA3K3_pLacGFP] - These are plasmid backbones optimized for use in Gibson cloning, with a variety of copy numbers and antibiotic resistances.

Magnetosome Toolkit

5. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590015 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. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K590016 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. [http://partsregistry.org/Part:BBa_K314100:Experience 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. [http://partsregistry.org/Part:pSB1A3:Experience 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. While pSB1A3 had a proper insert efficiency of 11%, the Gibson-optimized pGA1A3 vector was 99% efficient.

Improved Parts

8. [http://partsregistry.org/Part:BBa_K590059 BBa_K590059], [http://partsregistry.org/Part:BBa_K590060 BBa_K590060], [http://partsregistry.org/Part:BBa_K590061 BBa_K590061: LuxC, D, and E] (Cambridge, iGEM 2010) - Formerly part of the [http://partsregistry.org/Part:BBa_K325909 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. This was accomplished by the Alternate Aldehyde branch of the Alkane Production team.

All Submitted Parts