Lead Sensor Project
The BUGSS team has been working to cultivate a strain of E. Coli that produces metallothionein and RFP in the presence of heavy metals like lead. Adequate production of metallothionein protein can sequester lead in order to make the metal non-bioavailable. Again, the BUGSS team is building on earlier work on lead sequestration,utilizing some parts from the iGEM registry while producing and submitting new and composite parts. Sequences were found and edited for metallothionein production (BmtA), and the PbrR regulator. PpbrA promoters were connected to RFP and Bmta genes in order to regulate metallothienein and RFP production, inducing their generation in the presence of lead. In order for this new construct to work, the E. Coli's natural heavy metal resistance gene, Znta, must be deactivated. The team is utilizing a one step gene inactivation protocol using phage lambda red recombinase to recombine the gene with a kanamycin resistance insert.