Team:Cornell/project/wetlab/fungal toolkit

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<h6>Dry Lab</h6>
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<a href="https://2012.igem.org/Team:Cornell/project/drylab">How It Works</a>
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<a href="https://2012.igem.org/Team:Cornell/project/drylab/functional_requirements">Functional Requirements</a>
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<a href="https://2012.igem.org/Team:Cornell/project/drylab/components">Components</a>
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Modeling
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<h2 class="centered">How It Works</h2>
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<h2 class="centered">Fungal Toolkit</h2>
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<b>Scroll over each number to find out more about the components of our device!</b>
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<b>Scroll over each icon to find out more about the functional requirements for our device!</b>
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Water flows into our system, providing a sample to test water quality.
Water flows into our system, providing a sample to test water quality.
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Prevents foreign microbes from entering and contaminating our reactor.
Prevents foreign microbes from entering and contaminating our reactor.
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Move water and food to and from the reactor.
Move water and food to and from the reactor.
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Resist corrosion and set flow rates to the optimal level.
Resist corrosion and set flow rates to the optimal level.
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House bacteria that produce electrical current in response to toxins.
House bacteria that produce electrical current in response to toxins.
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Supply our bacteria with lactate to keep them alive.
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<h5>Continuous</h5>
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Pollution events can be hard to spot, making discrete testing inadequate as well as expensive. Monitoring water quality is a 24 hour job.
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Record bacterial current and transmit data wirelessly to the user.
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Keeps our device running.
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<h5>Field Deployable</h5>
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Industrial water monitoring is needed in remote and rugged terrain. In order to be applicable in these terrains, our device should be durable, water-proof. Being out in the field also means all food for bacteria and power for electronics must be provided by the device itself.  
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Extends the longevity of our device by providing extra power.
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Prevents our microbes from leaving the device.
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<h5>Remote</h5>
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To collect data in isolated locations, wireless communication is essential. This requires digital conversion of signals from our bacteria, so that it can be transmitted to the user and accessed online.  
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Water flows back to the source.
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Keeps the device safe from weather, rocks, and, of course, bears.
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Our device needs to output signal that is practical and intuitive. Electrical signal is easy to sense, process, and convert to digital signal. </div>
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In all of its glory.
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To monitor water quality in a truly continuous fashion, our device must be able to sustain itself without maintenance for long periods of time. Frequent maintenance is impractical.
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<h3>Protoplasting</h3>
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<h5>Background</h5>
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Transformation of plant and fungal cells is difficult due to their cell walls that block passage of foreign DNA. Protoplasting is the method by which the cells walls of plant and fungal cells are digested to produce cells without cell walls, called protoplasts. Through additional methods, such as electroporation and PEG transformation, DNA can be uptaken by the protoplasts and then regrown into cells containing specific genes.
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<h5>Method</h5>Usually this species is protoplasted using lywallzyme and novozyme. However, these enzymes are exclusively available in China. So, protoplasting was attempted with driselease and glucanex instead. After several attempts, we found that the enzymes are unable to digest the cell wall without also killing the cell. In addition, the Ganoderma mycelium was hard to pellet when centrifuging and made the protoplasting procedure difficult. Thus, <i>Cochliobolus heterostrophus</i> was used instead. Protoplasting was successful using driselease and glucanex and then transformed via PEG solution.   
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<h3>References</h3>
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Sun L, Cai H, Xu W, Hu Y, Gao Y, Lin Z (2001). Efficient Transformation of the Medicinal Mushroom Ganoderma lucidum. Plant Molecular Biology Reporter, 19, 383a-383j.
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Revision as of 19:00, 27 September 2013

Cornell University Genetically Engineered Machines

Fungal Toolkit

Scroll over each icon to find out more about the functional requirements for our device!
Inlet
Water flows into our system, providing a sample to test water quality.
Filter
Prevents foreign microbes from entering and contaminating our reactor.
Pumps
Move water and food to and from the reactor.
Piping and Calibration
Resist corrosion and set flow rates to the optimal level.
Reactors
House bacteria that produce electrical current in response to toxins.

Protoplasting

Background
Transformation of plant and fungal cells is difficult due to their cell walls that block passage of foreign DNA. Protoplasting is the method by which the cells walls of plant and fungal cells are digested to produce cells without cell walls, called protoplasts. Through additional methods, such as electroporation and PEG transformation, DNA can be uptaken by the protoplasts and then regrown into cells containing specific genes.
Method
Usually this species is protoplasted using lywallzyme and novozyme. However, these enzymes are exclusively available in China. So, protoplasting was attempted with driselease and glucanex instead. After several attempts, we found that the enzymes are unable to digest the cell wall without also killing the cell. In addition, the Ganoderma mycelium was hard to pellet when centrifuging and made the protoplasting procedure difficult. Thus, Cochliobolus heterostrophus was used instead. Protoplasting was successful using driselease and glucanex and then transformed via PEG solution.

References

Sun L, Cai H, Xu W, Hu Y, Gao Y, Lin Z (2001). Efficient Transformation of the Medicinal Mushroom Ganoderma lucidum. Plant Molecular Biology Reporter, 19, 383a-383j.

Retrieved from "http://2013.igem.org/Team:Cornell/project/wetlab/fungal_toolkit"