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Team Stritch: Metal-o-coli Removal of Heavy Metal from Contaminated Water

This project aims to understand synthetic biology and its real-world application in addressing environmental issues, specifically water contamination by heavy metals such as mercury and arsenic. The implementation strategy involves independent student research projects in collaboration with faculty, focusing on metal resistance systems in bacteria. The long-term goals include expanding the project to other heavy metal sequestration methods and offering a synthetic biology interdisciplinary course.

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Team Stritch: Metal-o-coli Removal of Heavy Metal from Contaminated Water

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  1. Team Stritch: Metal-o-coliRemoval of Heavy Metal from Contaminated Water SusmitaAcharya and Nighat Kokan (Chemistry/Biology Collaboration) Cardinal Stritch University Milwaukee, Wisconsin

  2. Project Goals/Ideas Application Based • Understanding synthetic biology and its real world application Our focus • Environmental problem and issues, specifically water contamination by heavy metals

  3. Project Proposal • Two projects that really interested us after reviewing the 2009 iGEM website were • Team UQ-Australia: Mercury sequestration using multicomponentoperon • Team Virginia: Arsenic sequestration for ground water decontamination • Team Stritch: Removal of heavy metals from contaminated water specifically mercury and arsenic

  4. Implementation Strategy • As independent student research projects in collaboration with biology and chemistry faculty • Students have 4 plus credits for independent research study • One credit • literature review, bioethical issues, and research proposal • Three credits • Working on synthetic biology projects • Writing their research paper • Poster and oral presentation at the Institution level and at regional conferences

  5. Metal Resistance Systems in Bacteria • Resistance genes on plasmids or chromosome • For example in mercury resistance • The system consists of • A metal transporter • A metal binding protein • Metal responsive promoter • Reduction mechanism, Hg (II) Hg (0)

  6. Project Design • Overview Diagram from: iGEM 2009 Team UQ Australia

  7. Project Design Plasmid 1 • Allows Hg (II) transportation into cell • Metallothionien protein aids intracellular accumulation • MerR is a transcriptional regulator • MerAreductase (missing) no efflux of Hg (0) Diagram from: iGEM 2009 Projects Team UQ Australia

  8. Project Design Plasmid 2 • MerR promoter linked to Ag43 • Ag43, a native protein to E.coli (E. coli K12) is removed from genome and reinserted on a plasmid which can be controlled by Hg (II) • When Ecoli cells express Ag 43 they clump • Measure using Spectrophotometer Diagram from: iGEM Team UQ Australia

  9. BioBricks Parts • BBa_I76101 (did not work ) • BBa_I728456 MerRT: Mercury inducible promoter +RBS • BBa_J63010 vector • Ag43 isolated from plasmid pKKJ143 • MerT and MerP from plasmid pSUTp • BBa_K205004 (MerT) • BBa_I728654 (mercury Induced RFP reporter)

  10. Long Term Goals • Expand present project to other heavy metal sequestration • To offer synthetic biology as a 2-3 credit project based interdisciplinary course • Design and construct BioBricks • Characterize DNA-based devices • Contribute to registry

  11. Acknowledgements • Our Thanks to • Malcolm Campbell • Laurie Heyer, Todd Eckdahl, and Jeff Poet • Davidson College

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