Team:Harvard/vectors

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<p>Our goal is the creation of an "iGarden" with the BioBrick standard. To approach this project, we used agrobacterium-mediated transformation to move our constructs from the bacterium e. coli to agrobacterium, and finally into the plant arabidopsis (and maybe in the future, 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>The process began with the creation of desired constructs in e. coli - 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, or <a href="https://2010.igem.org/Team:Harvard/fences">fence</a> mechanisms. We next built a set of <a href="https://2010.igem.org/Team:Harvard/vectors/vectors">plant vectors</a> compatible with the BioBrick standard and moved our parts from e. coli into these vectors, which allowed us to electroporate our parts into agrobacteria. The final step was to transform arabidopsis flowers with our engineered agrobacteria and select for plants containing our designed constructs.</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.