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.

Transcript

1. University of Wisconsin Biosensors Evolving Madison 2011
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.
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
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)