Revision as of 20:28, 5 May 2017

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.