UW-Madison's 2011 iGEM presentation
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University of Wisconsin Biosensors Evolving Madison 2011
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Our team of 8 undergrads spent a summer conceiving, designing, constructing, and testing a new system for sensing and reporting alternative fuel source concentrations. We were lucky enough to be invited to the world championship jamboree at MIT in Cambridge, MA, and this was our presentation for our 20 minute talk. Let me just say, I absolutely love how these slides turned out. However, it definitely makes more sense with our explanations, so check the whole talk out at the iGEM results website: http://2011.igem.org/files/video/Wisconsin-Madison_Championship.mp4
Transcript of UW-Madison's 2011 iGEM presentation
University of Wisconsin
BiosensorsEvolving
Madison2011
Meet theteam
Biofuels
BiosensorsTuning BiosensorsSharing
&UW-Madisonbiofuels
Biofuelscan help solvethe energy
crisis
But…high-tech
equipmentanalysis
requires
Biofuels
BiosensorsTuning BiosensorsSharing
How can iGEM teams detect biofuels?
biosensorsWhat are
and how can they help? Natural sensor
system+ Reporter
gene Biosensor
ethanolWe 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.
0.0% 0.5% 1.0% 1.5% 2.0% 2.5% 3.0% 3.5% 4.0% 4.5% 5.0%0
10000
20000
30000
40000
50000
60000
70000
80000
90000
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Uninduced
Induced
Concentration of Ethanol (v/v)
Flu
ore
sce
nce
(O
D n
orm
ali
ze
d)
Alkanesare sensed with one
Smits, T, et al. (2001). New alkane-responsive expression vectors for Escherichia coli and Pseudomonas. Plasmid 46, 16-24.
How can we make our sensors
better?
Biofuels
Biosensors
Tuning BiosensorsSharing
How can iGEM teams detect biofuels?
How can we improve our biosensors?
Directedwith K634007
RFPsacB
kanR
evolution
RFPsacB
kanR
RFPsacB
kanR
0 3 6 9 12 15 18 210
0.1
0.2
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0.8
Time(h)
OD
60
0
Sucrosesensitivity?
RFPsacB
kanR
Sucrosemutants
0 5 10 15 20 25
-0.1
0
0.1
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0.8
Initial K634007 culture
Post-counter selection cells
Plasmid of post-counter selection cells in new DH10B
Time(h)
OD
600
0 500 1000 1500 2000 2500 3000 3500 4000 4500 50000
0.1
0.2
0.3
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0.5
0.6
0.7
0.8
non-inducedinduced
Kanamycin concentration (mM)
OD
600
RFPsacB
kanR
Kanamycinresistance
Evolveanything!
Biofuels
Biosensors
Tuning Biosensors
Sharing
How can iGEM teams detect biofuels?
How can we improve our biosensors?
Can directed evolution help iGEM teams?
DirectedReporter A1
Reporter A2
Reporter A3Evolution
Now what?I have all these mutants…
Introducing
mutant libraries
Introducing
mutant libraries
Introducing
mutant libraries
Mutagenesis Tree
Better organizationequals
better registry
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.
Increasingawareness
Increasingawareness
Increasingawareness
@LabBadger
TremendousThe College of EngineeringThe students of the Pfleger labThe GLBRC, NSEC, and MRSECThe UW-Madison REU programiGEM HQ and the registryJohn Greenler and the GLBRC
thanks to:
Questions?
(I know you have some)
