bio detection of anoxia in lake xochimilco using modified
TRANSCRIPT
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Bio detection of anoxia in Lake Xochimilco using modified Escherichia coli
by GenetiX
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Instituto Tecnológico y de Estudios Superiores de Monterrey, Campus Ciudad de México
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Our Team
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Background Xochimilco:
• Source of water for the Metropolitan Area
• Highly touristic and agricultural element
• World Heritage Site
• Local endemic species in danger of extinction
• Highly polluted with indutrial residues
• Critical levels of dO2 below minimun requirements
• Artificial eutrophication
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What is Eutrophication?
Dissolved Oxygen
Less than 3 mg/L
Movement of water
Temperature
Salinity
Pressure
Photosynthesis
Respiration
Eutrophication
Factors
Water
Xochimilco
6 mg/L
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Eutrophication
Increased Photosynthesis
Nutrient Fertilizers
Nitrogen and Phosphorus
Blocking of light to littoral zone
Microbial decomposition
Depletion of dissolved oxygen
Hypoxia Less than 2.8mg/L
Indicators Chlorophyl
Parameter Oligotrophic Eutrophic
Average total phosphorus 8.0 84.4
Average total nitrogen 661 1875
Average Chlorophyll a 1.7 14.3
Chlorophyll a. peak concentration 4.2 42.6
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Objective
Detection of anormal ferric and anoxia/hipoxia
levels on water samples from the Xochimilco Lake by
using biosensors
O2Prom
2 ppm>[O2]
2 ppm<[O2]
Fe Prom
0.5ppm>[Fe]
0.5ppm<[Fe]
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Topic
Biosensors:
• Fast response in time
• Fast and continuous measurement
• High specificity (shape-specific
recognition)
• Simplicity in its use
• Capability of measuring
concentrations ranging from 10-18 to
10-19 M, so we need low sample
requirements
• Real time measurements
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Justification
• Acceptable levels of disolved iron in water: 0.5
ppm
• Iron promoter minimal iron dissolution
concentration: 1 ppm
• Oxygen Promoter Works in dO2 concentrations
over 0.2 ppm
• dO2 levels in Xochimilco vary around 3 ppm
• Minimal requirement of dO2 for wild-life
sustainance 6 ppm
• Detection of quantitative values of GFP and RFP
by Flow Cytometry
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Our Biosensor
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Methodology & Results
A B
+ Bacteria
Transformation
20 hours
37° C
A B A B
A
B
Concentrated DNA
A
B
Transformed Bacteria
Heat Shock
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Methodology & Results A B
Concentrated DNA
Check +enzymes
+Enzymes
Expected Pattern
Agarose Gel
Cut Bands A
B Ligation
A B
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Methodology & Results
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Oxygen Promoter + G.F.P
Iron Promoter + G.F.P
Oxygen Promoter + mR.F.P.
EcoRI Pstl
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Discussions
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Conclusions
Construction of biosensors was not completed
due to what appear to be flawed ligations
and/or low efficiency competent cells.
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Future Work
Completion of the biosensors
Proof of concept growing bacteria in absence
and presence of stimuli
Quantitative measurement of disolved O₂ and
Fe in various concentrations for construction of
a photoluminescent scale.
Application of biosensors to a range of samples
from specific points distributed all along
Xochimilco lake.
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Posada Tec 2013
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Travesuras 2014
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Synthetic Biology Conference
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Thanks
ITESM CCM
Hidrosina
GoNext Project
Tecnologico de Monterrey campus Mty.
Nautilus
Tolerantum
QFB. Isabel Coria
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Special Thanks
Luis Vaca Dominguez M.D. Ph. D.
Alicia Sampieri García Ph. D.
M.Sc. Lamberto Álvarez-Palacios