“Light to live, live to light”2012.igem.org/files/presentation/Valencia.pdf · Submitted parts:...
Transcript of “Light to live, live to light”2012.igem.org/files/presentation/Valencia.pdf · Submitted parts:...
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“Light to live, live to light” Valencia 2012
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PHOTOSYNTHETIC
BIOLAMP
The project:
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The project:
AUTOSUFFICIENT
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The project:
SOLAR POWER
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The project:
AUTOREGULATED
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The project:
ENERGY EFFICIENT
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The project:
COLD LIGHT
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The project:
NON ELECTRIC
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The project:
ENVIRONMENTALLY FRIENDLY
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The project:
NO FUEL INPUT
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The project:
CO2 SINK
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The project:
SYNTHETIC ECOLOGY
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The project:
ARTIFICIAL TROPHIC
SYMBIOSIS
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INTERSPECIES CELL-TALK
The project:
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Synechococcus elongatus Aliivibrio fischeri
Synergy can be the
greatest power:
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BATTERY AND SWITCH
BIOLAMP
Energy Signal
molecule
The
idea:
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Aliivibrio fischeri
Yummy!
suc: sucrose
PpsbAI λ P LuxI
cscB
LuxR LuxICDABE
S. elongatus cscB
LuxR LuxR
LuxR
cI repressor
Lac P
BATTERY AND SWITCH
BIOLAMP
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Aliivibrio fischeri
AHL
Yummy!
suc: sucrose
psbAI cI repressor λ P
cscB
LuxR LuxICDABE
S. elongatus cscB
AHL
AHL
LuxR
AHL
LuxR
LuxR
LuxI
Lac P
BATTERY AND SWITCH
BIOLAMP
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Integrate both organisms with maximum
efficiency
•Continuous culture •Separate modules
Photosynthesis and Export
Synechococcus elongatus
Biolamp Aliivibrio
fischeri
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Designing the common
medium Meet requirements of both organisms:
Synechococcus elongatus Aliivibrio fischeri
High pH High pH
Saline medium Saline medium
Micronutrients C, N organic sources
Synechococcus strain growing in BG-11M enriched with 250mM of salt.
Aliivibrio strain growing in BG-11M enriched with salt and organic sources.
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Submitted parts
Overview
Human Practices
Achievements
Modeling
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Modelling the
parameters Find the optimal bioreactor design parameters:
• Volumes of the cultures
• Cell densities
In order to optimize:
• Enough light emission
• Stable day/night switch
• Sucrose balance to keep our system autonomous
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Metabolic network with 898 reactions + 803 metabolites.
Synechococcus metabolic model + 2 extra reactions: - sucrose export - AHL
LIGHT CO2
+ BIOLOGICAL CONSTRAINTS
Sucrose AHL Growth
Synechococcus
model
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Assay production of AHL and sucrose
0.0221mmol/ gDCW·h
Synechococcus
model Growth: exponential rate
(mmol/ gDCW·h)
(mmol/ gDCW·h)
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Belta et al, 2001
9 differential equations to predict light emission:
x7´= (Vc[Ai] + Vv.X7 )/Vt
Aliivibrio QS
model
where:
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Solar Light CO2
Synechococcus Metabolic network
Aliivibrio Bioluminescence model
AHL AHL SUC SUC
AHL
AHL
Luminescence
SUC
Synergic
model
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Submitted parts
Overview
Modeling
Achievements
Human Practices
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YES 56% NO
14%
N/A 25%
Population Poll
Human practices:
Synth(ethic) Biology Blog http://igemvalencia2012.wordpress.com/
Kids lectures
Seminars
Indifferent 5%
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Overview
Modeling
Human Practices
Achievements
Submitted parts
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Submitted parts:
Our submitted part in the pSB1C3 standard BioBrick
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Submitted parts:
Testing the part:
Dark (h)
Light (h)
Measure starts at
PpsbAI::luxAB 1 1 Light cycle Dark cycle
PpsbAI::luxAB 8 16 Light cycle Dark cycle
PpsbAI::luxAB 16 8 Light cycle Dark cycle
PpsbAI::luxAB 12 12 Light cycle Dark cycle
PpsbAI::luxAB 0 24 Light cycle Dark cycle
Negative control (WT)
Same Same Same Same At three different light intensities
PpsbAI::luxCDABE fusion vector
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Submitted parts
Overview
Modeling
Human Practices
Achievements
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Achievements:
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Designed a self-sufficient photosynthetically-powered biolamp.
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Studied an artificial consortium between two marine organisms.
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Built an effective model for the different biolamp processes.
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Transformed Synechococcus elongatus PCC7942 to characterize the psbAI promoter.
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Submitted a well characterized part to the Registry of Standard Biological Parts.
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Opened a bilingual weblog about synthetic biology, focusing on divulgation.
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Established collaborations with others iGEM teams: Uppsala, UC London, Copenhagen, Chile.
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Tested and characterized new culture medium for growing cyanobacteria in our lab conditions.
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Acknowledgments
• Daniel Ducat
• Susan Golden
• Javier Espinosa, from Fisiology, Genetic and Microbiology departments of University of Alicante
• Dr. Carratalá
• Marisa Salvador, from the Molecular Biology Department of University of Valencia
• Joaquín Moreno, from Molecular Biology Department of University of Valencia
• Diego Orzáez, from IBMCP(UPV-CSIC)
• Borja Martinez-Clavel Vallés
• Marta Chillerón Rueda
• Raul Gancedo Sabater