Team:iTesla-SoundBio/Attributions

The production of PCBs has been banned in the United States because of possible impact on animal and human health, yet the harmful molecules still contaminate our waters today. Our project involves transferring genes responsible for producing PCB-dechlorinating enzymes from the anaerobic D. mccartyi bacterium, in which they're naturally found, into E.coli, an easier-to-work-with bacterium.

We will be transferring three different genetic variants of the PCB dechlorinating enzyme and will face many challenges in producing the working enzymes. The enzymes probably use norpseudo-B12 as a cofactor, a compound E. coli doesn't naturally create. We are currently researching ways to introduce this into the environment of the E.coli expressing the proteins. The working enzymes also contain iron-sulfer clusters, which will be destroyed upon exposure to oxygen, as the original organism is anaerobic. We are currently looking into different and easy ways to grow E.coli anaerobically and ways to combat this issue. D. mccartyi's membrane is also archea-like and dissimilar to E.coli's membrane, which may cause complications with expression and localization. The enzymes are also transmembrane proteins, which may pose problems.

We will be contacting other labs with higher biosafety levels due to the toxicity of PCBs, to test out our system. We are currently working in lab to transfer our synthesized genetic circuits to E.coli bacteria while we research ways to solve the potential problems listed above. We have also included His tags in our genetic circuits and have partnered with the Baker Lab at the University of Washington to analyze our PCB dechlorinating enzymes in the future.

Attributions

iTesla IGEM has been through a lot in its first year, and we would like to thank everybody who contributed no matter big or small.

SoundBio Lab

Special thanks to SoundBio Lab in Seattle for hosting our team throughout the year! SoundBio provided not only space for us to work at but also mentors to guide us through the season.

  • Zach Mueller
  • Alan Bohn

University of Washington IGEM Team

Special thanks to UW IGEM, they allowed us to use their nanodrop for quantifying yields!

  • Chris Choe

University of Washington

  • Joel Baker
    • Special thanks to Mr. Baker for visting and explaining to our team how to design our experiment for quantifying expression.
    • http://directory.tacoma.uw.edu/employee/jebaker

Gene Wiz

Special thanks to Gene Wiz for sequencing our sanger samples[samples were prepared by iTesla IGEM members]

New England Biolabs

Special thanks to New England Biolabs for donating supplies.

Integrated DNA Technologies

Special thanks to Integrated DNA Technologies for donating kilobases of synthesis.

Old Redmond Firehouse

Special thanks to the Old Redmond Firehouse for hosting our team and providing us a place to meet.