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Cornell iGEM

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Wet Lab

Future Work

In the future, we hope to continue working with the merT, merP, CBP4, and nixA heavy metal transport genes by incorporating them upstream of the GST and CRS5 metallothionein sequestration genes. Once each heavy metal transport gene is combined with the metallothionein gene, we can transform the high copy bacterial plasmid into E. coli. We will then be able to conduct a series of growth assays between our engineered bacteria and E. coli in the presence of heavy metal contaminated water.

We also hope to continue working on synthesizing a reporter system. In order to detect the saturation of metallothionein sequestering cultures, we plan on using amilCP behind the nickel/cobalt activated promoter Prcn and the mercury activated promoter PmerT. It would be useful to place amilCP behind a lead activated promoter. This system should be incorporated into the BioBrick backbone and transformed into E. coli reporter cultures. These would theoretically be placed into a second hollow fiber reactor that would be connected downstream to the transporter-metallothionein hollow fiber reactor. Effluent water carrying unsequestered metal ions would induce the reporter culture to express amilCP, producing a gradient of blue. We can then test water samples with different heavy metal concentrations to correlate effluent levels against the cultures’ color gradient.