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eau d ’ e coli igem 2006: mit

eau d ’ e coli igem 2006: mit. The Undergrads: Kate Broadbent Biological Engineering Andre Green Biology / Physics Stephen Payne Biological Engineering Veena Venkatachalam Chemistry / Physics Boyuan Zhu Electrical Engineering and Computer Science The Grad Students: Barry Canton

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eau d ’ e coli igem 2006: mit

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  1. eau d’e coliigem 2006: mit

  2. The Undergrads: Kate Broadbent Biological Engineering Andre Green Biology / Physics Stephen Payne Biological Engineering Veena Venkatachalam Chemistry / Physics Boyuan Zhu Electrical Engineering and Computer Science The Grad Students: Barry Canton Austin Che Jason Kelly Reshma Shetty Samantha Sutton The Faculty: Drew Endy Tom Knight meet the team

  3. motivations eau d’e coli mit igem 2006

  4. background A. thaliana O. basilicum A. majus C. breweri S. cerevisiae JMT jasmine CCMT cinnamon BAMT fresh / floral ATF1 banana BSMT wintergreen

  5. background O. basilicum A. majus C. breweri S. cerevisiae CCMT cinnamon BAMT fresh / floral ATF1 banana BSMT wintergreen

  6. background A. majus C. breweri S. cerevisiae BAMT fresh / floral ATF1 banana BSMT wintergreen

  7. background C. breweri S. cerevisiae ATF1 banana BSMT wintergreen

  8. timeline:initial planning initial planning • scent production • self-generating scent • production regulation • eliminating indole

  9. timeline:scent production initial planning • scent production • self-generating scent • production regulation • eliminating indole

  10. timeline:self-generating scent initial planning • scent production • self-generating scent • production regulation • eliminating indole

  11. timeline:production regulation initial planning • scent production • self-generating scent • production regulation • eliminating indole

  12. timeline:indole elimination initial planning • scent production • self-generating scent • production regulation • eliminating indole

  13. wintergreen generating device

  14. E. coli + precursor (SA) + WGD E. coli methyl salicylate MW = 152 no methyl salicylate [no peak at 9.1 minutes] device generates wintergreen

  15. banana generating device

  16. precursor biosynthesis

  17. precursor biosynthesis

  18. goal: the regulated system

  19. growth dependent regulation • osmY: active in stationary phase & under high osmotic pressure conditions osmY osmY + inverter

  20. regulation is growth dependent

  21. regulation is growth dependent

  22. regulated system: exponential

  23. regulated system: stationary

  24. regulated system

  25. wild type E. coli chassis: tnaA5-E. coli Indole Peak at 10 minutes MW = 117 No indole [no peak at 10 minutes] chassis (tnaA5-) does not produce indole

  26. final system

  27. smell test results

  28. future applications • Improve the workplace environment for microbiologists working with E. coli • Port our system to bacterial species involved in bioremediation. • Implement our system in bacteria responsible for bad human odor in the mouth, armpits and feet. • Implement our system in yeast used in bread and beer production

  29. conclusion • Designed, built, and tested a synthetic biological system over the course of summer • Engineered synthetic scent systems in E. coli • Regulated via transcription control devices

  30. acknowledgments • Natalia Dudareva, Purdue University: gifts of expression vectors encoding BAMT, SAMT and BSMT enzymes. • Eran Pichersky, University of Michigan: suggestion of eliminating indole from Escherichia coli to mitigate the natural "bad" smell. • Mary Berlyn, CGSC, The Coli Genetic Stock Center: Esherichia coli strain YYC912. • Brian Cook, MIT: valuable discussions. • Cornelia Reimmann, University of Lausanne: gift of an expression vector carrying the pchBA coding region. • Dieter Haas, University of Lausanne: gift of the Pseudomonas fluorescens strain CHA0. • Peter Bakker, Utrecht University, The Netherlands: gift of an expression vector carrying the pmsCEAB coding region, as well as, gift of the Pseudomonas fluorescens strain WCS374. • Herbert P. Schweizer, Colorado State University: gift of the pUCP22 Esherichia coli to Pseudomonas shuttle vector. • Pamela Silver, Harvard Medical School: gift of yeast BioBricks vector.

  31. questions and answers

  32. growth curves

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