UW-Madison iGEM 2011 World Championship

6e03c3155f6e4a02506741c2023448d2?s=47 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.

6e03c3155f6e4a02506741c2023448d2?s=128

ewalters

January 24, 2012
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Transcript

  1. 2.
  2. 8.

    biosensors What are and how can they help ? Natural

    sensor system + Reporter gene Biosensor
  3. 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.
  4. 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
  5. 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.
  6. 12.
  7. 13.
  8. 15.
  9. 17.
  10. 18.
  11. 19.
  12. 20.
  13. 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?
  14. 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
  15. 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
  16. 26.

    Biofuels Biosensors Tuning Biosensors Sharing How can iGEM teams detect

    biofuels? How can we improve our biosensors? Can directed evolution help iGEM teams?
  17. 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.
  18. 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: