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UW-Madison iGEM 2011 World Championship

ewalters
January 24, 2012

UW-Madison iGEM 2011 World Championship

We spent the whole summer developing and improving E. coli strains to sense alternative fuels and then the morning of 11/6/11 showing them off to the world.

ewalters

January 24, 2012
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  1. University of Wisconsin Biosensors Evolving Madison 2011

  2. None
  3. Biofuels Biosensors Tuning Biosensors Sharing

  4. & UW-Madison biofuels

  5. Biofuels can help solve the energy crisis

  6. But… high-tech equipment analysis requires

  7. Biofuels Biosensors Tuning Biosensors Sharing How can iGEM teams detect

    biofuels?
  8. biosensors What are and how can they help ? Natural

    sensor system + Reporter gene Biosensor
  9. ethanol We can sense with 2 components Vangai, A, et

    al. (2009). Analysis of the promoter activities of the genes encoding three quinoprotein alcohol dehydrogenases in Pseudomonas putida HK5. Microbiology 155, 594-603.
  10. 0 10 20 30 40 50 60 70 80 90

    100 0.0% 1.0% 2.0% 3.0% 4.0% 5.0% Fluorescence (OD normalized) Concentration of Ethanol (v/v) Uninduced Induced
  11. Alkanes are sensed with one Smits, T, et al. (2001).

    New alkane-responsive expression vectors for Escherichia coli and Pseudomonas. Plasmid 46, 16-24.
  12. None
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  14. How can we make our sensors better?

  15. Biofuels Biosensors Tuning Biosensors Sharing How can iGEM teams detect

    biofuels? How can we improve our biosensors?
  16. Directed with K634007 RFP sacB kanR evolution

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  21. RFP sacB kanR

  22. RFP sacB kanR 0 0.1 0.2 0.3 0.4 0.5 0.6

    0.7 0.8 0 3 6 9 12 15 18 21 OD600 Time(h) 0% Sucrose 0.01% 0.03% 0.05% 0.10% 0.50% 1.00% 2.50% 5.00% Sucrose sensitivity?
  23. RFP sacB kanR Sucrose mutants -0.1 0 0.1 0.2 0.3

    0.4 0.5 0.6 0.7 0.8 0 5 10 15 20 25 OD600 Time(h) Initial K634007 culture Post-counter selection cells Plasmid of post-counter selection cells in new DH10B
  24. 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0

    1 2 3 4 5 OD600 Kanamycin concentration (mM) non-induced induced RFP sacB kanR Kanamycin resistance
  25. Evolve anything!

  26. Biofuels Biosensors Tuning Biosensors Sharing How can iGEM teams detect

    biofuels? How can we improve our biosensors? Can directed evolution help iGEM teams?
  27. Directed Reporter A1 Reporter A2 Reporter A3 Evolution

  28. Now what? I have all these mutants…

  29. Introducing mutant libraries

  30. Introducing mutant libraries

  31. Introducing mutant libraries

  32. Mutagenesis Tree

  33. Better organization equals better registry

  34. Biofuels Biosensors Tuning Biosensors Sharing How can iGEM teams detect

    biofuels? How can we improve our biosensors? Powerful in theory, what comes next? New tools require new technologies.
  35. Increasing awareness

  36. Increasing awareness

  37. Increasing awareness @LabBadger

  38. Tremendous The College of Engineering The students of the Pfleger

    lab The GLBRC, NSEC, and MRSEC The UW-Madison REU program iGEM HQ and the registry John Greenler and the GLBRC thanks to:
  39. Questions? (I know you have some)