Difference between revisions of "Team:UNIK Copenhagen/Workshop"

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<h1>More to come shortly...</h1>
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<h1>Presentations</h1>
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<font size="4">Human Exploration and Space Flight</font> <br>
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<font size="3">Kiruna, Sweden August 15th 2015 </font>
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<img src="https://static.igem.org/mediawiki/2015/1/12/UNIK_Copenhagen_Kirunapresentation.jpg" width=72%></div>
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<img src="https://static.igem.org/mediawiki/2015/b/bf/UNIK_Copenhagen_Spacemossground.jpg" width=40%></div>
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<p>Outside the scope of the competition we will insert the DNA gene containing YFP and an antifreeze protein gene and a resveratrol gene. When this is transformed into the moss we should have an output of moss that is able to produce the antifreeze protein on its own, or produce resveratrol. The moss will also glow yellow due to the YFP when it is producing either of the components, which acts as a proof of concept. </p>
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<font size="4">Technical description</font>
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<p>We will make a stable and transient transformation of moss, Psychometrilla patens. We will make two linear gene constructs for stable transformation in moss. One construct is going to contain a gene encoding an antifreeze protein and the other construct will contain a stilbene synthase-gene (STS), making the final enzyme in the resveratrol pathway. Both constructs contain regions homologous to the moss genome, so they will integrate via homologous recombination in moss. The constructs furthermore contains the ZmUbi-promoter, resistance gene and YFP, to confirm transformation. We will also make a transient moss transformation with a vector containing the ZmUbi-promoter followed by YFP, to confirm the function of the promoter in moss. </p>
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Moss growing facility</div>
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<p>Due to time constraints, we will also transform tobacco and <i>E.coli</i>. We will transform tobacco with the STS-gene, using USER-cloning and Agrobacterium tumefaciens transformation, in order to quickly confirm the functionality of the gene (moss takes a long time to grow!) by measuring the presence of resveratrol with Liquid chromatography-mass spectrometry.
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Furthermore we will transform E. coli with the antifreeze gene which originate from the spruce budworm, to quickly see whether the gene has an effect on temperature sensitivity. We will perform experiments at low temperatures to measure the growth of the bacteria when exposed to cold. We will also make a his-tag version of the antrifreezeprotein, to confirm the presence of the protein by western blotting.</p>
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    <img src="https://static.igem.org/mediawiki/2015/b/b9/UNIK_copenhagen_mossinhand.jpg" height="512" width="341.25" />
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    <p>One of our petri dishes with moss</p>
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    <img class="middle-img" src="https://static.igem.org/mediawiki/2015/8/89/UNIK_copenhagen_mossfacility.jpg"/ height="512" width="341.25" />
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    <p>Moss growing facility</p>
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Revision as of 13:46, 19 August 2015


Presentations

Human Exploration and Space Flight
Kiruna, Sweden August 15th 2015







Outside the scope of the competition we will insert the DNA gene containing YFP and an antifreeze protein gene and a resveratrol gene. When this is transformed into the moss we should have an output of moss that is able to produce the antifreeze protein on its own, or produce resveratrol. The moss will also glow yellow due to the YFP when it is producing either of the components, which acts as a proof of concept.



Technical description

We will make a stable and transient transformation of moss, Psychometrilla patens. We will make two linear gene constructs for stable transformation in moss. One construct is going to contain a gene encoding an antifreeze protein and the other construct will contain a stilbene synthase-gene (STS), making the final enzyme in the resveratrol pathway. Both constructs contain regions homologous to the moss genome, so they will integrate via homologous recombination in moss. The constructs furthermore contains the ZmUbi-promoter, resistance gene and YFP, to confirm transformation. We will also make a transient moss transformation with a vector containing the ZmUbi-promoter followed by YFP, to confirm the function of the promoter in moss.



Moss growing facility






Due to time constraints, we will also transform tobacco and E.coli. We will transform tobacco with the STS-gene, using USER-cloning and Agrobacterium tumefaciens transformation, in order to quickly confirm the functionality of the gene (moss takes a long time to grow!) by measuring the presence of resveratrol with Liquid chromatography-mass spectrometry. Furthermore we will transform E. coli with the antifreeze gene which originate from the spruce budworm, to quickly see whether the gene has an effect on temperature sensitivity. We will perform experiments at low temperatures to measure the growth of the bacteria when exposed to cold. We will also make a his-tag version of the antrifreezeprotein, to confirm the presence of the protein by western blotting.



One of our petri dishes with moss

Moss growing facility