Team:Cornell/project/wetlab/fungal toolkit

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<h2 class="centered">Fungal Toolkit</h2>
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<h3>Protoplasting</h3>
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<b>Scroll over each icon to find out more about the components of our toolkit!</b>
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<h5>Background</h5>
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Transformation of plant and fungal cells is difficult due to their cell walls which blocks 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>
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Protoplasting was first attempted on <i>Ganoderma lucidum</i>. Usually this species is protoplasted using lywallzyme and novozyme. However, these enzymes are exclusievly available in China and thus protplasting was attempted with driselease and glucanex. After several attempts it was concluded that the enzymes were 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 ____</i> was used instead. Protoplasting was sucessful using driselease and glucanex and then transformed via PEG solution.    
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<h5>Regulatory Elements</h5>
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Provide modular protein expression controls for genetic constructs.
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<h5>Selectable Markers</h5>
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Allow for the selection of desired fungal transformants.
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<h5>Characterization</h5>
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Quantifies gene expression levels using fluorescent proteins.
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<h5>Homologous Constructs</h5>
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Enable integration of genetic constructs into specific regions of fungal genomes.
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<h5>Biosafety</h5>
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Implements measures to prevent horizontal gene transfer and induce a kill switch.
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Latest revision as of 13:01, 12 October 2013

Cornell University Genetically Engineered Machines

Fungal Toolkit

Scroll over each icon to find out more about the components of our toolkit!
Regulatory Elements
Provide modular protein expression controls for genetic constructs.
Selectable Markers
Allow for the selection of desired fungal transformants.
Characterization
Quantifies gene expression levels using fluorescent proteins.
Homologous Constructs
Enable integration of genetic constructs into specific regions of fungal genomes.
Biosafety
Implements measures to prevent horizontal gene transfer and induce a kill switch.

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