Team:Cornell/project/wetlab/assembly

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Shewanella oneidensis MR-1’s metal reduction pathway can be likened to a simple switch analogy. Without MtrB, no extracellular transfer of electrons is possible, and therefore no current is produced. However, when reintroduced, MtrB closes the switch therefore allowing extracellular reduction and current generation.

To sensitize Shewanella’s metal reduction pathway to our analytes, we decided to use a complementation strategy. By using the Shewanella MtrB knockout strain, JG 700, which was graciously provided by Professor Jeffery Gralnick from the University of Minnesota, we are able to reintroduce MtrB on a plasmid under the control of inducible promoters sensitive to the analytes we want to detect. Thus, MtrB—and therefore current—should only be produced in the presence of analyte.

One of the greatest strengths of this approach is its modularity; by simply switching out the sensing region on the plasmid, we can sensitize MtrB production to any analyte for which genetic parts exist.