Team:Harvard/vectors

From 2010.igem.org

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<p>Our goal is the creation of an "iGarden" with the biobrick standard and in order for such a concept to work, we must get our constructs into plants. We used agrobacterium-mediated transformation, first in arabidopsis, then in strawberries. </p>
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<p>Our aim in creating the iGarden was to create a framework for plant engineering following the BioBrick standard. We began by modifying a set of <a href="https://2010.igem.org/Team:Harvard/vectors/vectors">vectors</a> designed to facilitate agrobacterium-mediated transformation to be compatible with the BioBrick standard, and creating a collection of constructs to be expressed in plants - such as <a href="https://2010.igem.org/Team:Harvard/flavor">flavor</a> elements, <a href="https://2010.igem.org/Team:Harvard/allergy">allergen</a> knockdown, <a href="https://2010.igem.org/Team:Harvard/color">pigment</a> accumulation, and <a href="https://2010.igem.org/Team:Harvard/fences">containment</a> mechanisms. We built all constructs in <i>E. coli</i>, and once complete, cloned them into the BioBrick agrobacterium vectors. We then transformed agrobacteria with our constructs via electroporation, and subsequently transformed <i>Arabidopsis</i> seeds by dipping flowers into the transgenic agrobacteria. In our final step, we harvested the transformed seeds and screened them for our designed constructs using antibiotic selection.</p>
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<p>To do so, we created a set of <a href="vectors.html">vectors</a> compatible with the biobrick standard. To express our constucts - such as <a href="flavor">flavor</a> elements, <a href="2010.igem.org/Team:Harvard/allergy">allergen</a> knockdown, <a href="2010.igem.org/Team:Harvard/color">pigment</a> accumulation, or <a href="2010.igem.org/Team:Harvard/fence">fence</a> mechanisms, we cloned the constucts into the agrobacterium vectors with E. coli as the chassis, then transformed into agrobacteria via electroporation, then innoculated our plants with the contruct containing agrobacteria. </p>
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<img src="https://static.igem.org/mediawiki/2010/0/03/Harvard2010roadmap.png" width="600px">
<img src="https://static.igem.org/mediawiki/2010/0/03/Harvard2010roadmap.png" width="600px">

Latest revision as of 23:45, 26 October 2010



abstract

Our aim in creating the iGarden was to create a framework for plant engineering following the BioBrick standard. We began by modifying a set of vectors designed to facilitate agrobacterium-mediated transformation to be compatible with the BioBrick standard, and creating a collection of constructs to be expressed in plants - such as flavor elements, allergen knockdown, pigment accumulation, and containment mechanisms. We built all constructs in E. coli, and once complete, cloned them into the BioBrick agrobacterium vectors. We then transformed agrobacteria with our constructs via electroporation, and subsequently transformed Arabidopsis seeds by dipping flowers into the transgenic agrobacteria. In our final step, we harvested the transformed seeds and screened them for our designed constructs using antibiotic selection.