Team:Harvard/fences/results

From 2010.igem.org

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<h1>results</h1>
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<h2>induction of barnase (death gene) reduces cell growth</h2>
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<b>Barnase Growth Control</b>
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<p>The fatal gene is the key element of the genetic fence, without which no containment will result. To confirm the ability of barnase to contain plants outside the genetic fence, we characterized an inducible barnase system created by the Berkeley 2008 iGEM team.</p>
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<p>In this construct, arabinose induction results in Barnase expression. Because of the presence of low background levels of the inhibitor Barstar, low levels of Barnase expression will not immediately kill the cells. Higher and higher levels of Barnase expression results in lower and lower rates of growth in the cells, affirming the principle of Barnase-based growth control in the genetic fence.</p>
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<div>Barnase Growth Control in E.Coli &nbsp; <a href="http://openwetware.org/images/2/2b/M4-growth_curves-10-4-10.jpg" style="font-size:12px">click to enlarge</a></div><hr/>
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<p>This barnase system works by expressing background levels of Barstar with Barnase inducible such that it will overwhelm Barstar when induced.
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<p>We characterized the activity of Barnase on an inducible plasmid constructed by <a href="https://2007.igem.org/BerkiGEM2007Present5">UC Berkeley for iGEM 2007</a> (parts BBa_I716408C and BBa_I716211). This construct works by expressing background levels of Barstar with in the presence of an inducible Barnase. When induced, Barnase will overwhelm Barstar. Higher levels of Barnase expression resulted in lower rates of growth in the bacteria, affirming the principle of Barnase-based growth control for the genetic fence, and confirming the results from Berkeley 2008. We characterized the growth repression of Barnase under a range of arabinose inducer concentrations.<br><br>
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A plate reader assay demonstrated the ability of Barnase to inhibit growth in Bacteria, confirming the results from Berkeley 2008.</p>
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Our results show that expression of Barnase is effective in reducing cell growth, suggesting that Barnase will enable the genetic fence to prevent growth of iGarden plants outside of their designated areas.
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<div><b>barnase growth control in <i>E. Coli</i></b> &nbsp; <a href="http://openwetware.org/images/2/2b/M4-growth_curves-10-4-10.jpg" style="font-size:12px">[click to enlarge]</a></div><hr/>
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<img src="http://openwetware.org/images/2/2b/M4-growth_curves-10-4-10.jpg" width="600px" border=0>
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</a> </div>
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<b>Barstar Inhibition of Barnase</b>
 
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Latest revision as of 18:09, 27 October 2010



induction of barnase (death gene) reduces cell growth

We characterized the activity of Barnase on an inducible plasmid constructed by UC Berkeley for iGEM 2007 (parts BBa_I716408C and BBa_I716211). This construct works by expressing background levels of Barstar with in the presence of an inducible Barnase. When induced, Barnase will overwhelm Barstar. Higher levels of Barnase expression resulted in lower rates of growth in the bacteria, affirming the principle of Barnase-based growth control for the genetic fence, and confirming the results from Berkeley 2008. We characterized the growth repression of Barnase under a range of arabinose inducer concentrations.

Our results show that expression of Barnase is effective in reducing cell growth, suggesting that Barnase will enable the genetic fence to prevent growth of iGarden plants outside of their designated areas.


barnase growth control in E. Coli   [click to enlarge]