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Team:Edinburgh/Project/Future
From 2010.igem.org
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<a name="Future" id="Future"></a><h2>Future Work: Sequential addition</h2> | <a name="Future" id="Future"></a><h2>Future Work: Sequential addition</h2> | ||
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| - | <p>One of the future expansions of BRIDGE that we have discussed involves using it to directly introduce genes in the genome next to each other without using the BioBrick method before-hand. For example, if you wanted to insert four genes with the steps described in the protocol section, it would take eight steps. If you do this with the method shown in Figure 1 below, it would only take four steps.</p><br> | + | <p>One of the future expansions of <a href="https://2010.igem.org/Team:Edinburgh/Project/Protocol">the BRIDGE protocol</a> that we have discussed involves using it to directly introduce genes in the genome next to each other without using the BioBrick method before-hand. For example, if you wanted to insert four genes with the steps described in the protocol section, it would take eight steps. If you do this with the method shown in <a href="https://static.igem.org/mediawiki/2010/7/71/Ed10-SequentialBridge.JPG">Figure 1</a> below, it would only take four steps.</p><br> |
<center><p><img src="https://static.igem.org/mediawiki/2010/7/71/Ed10-SequentialBridge.JPG" width="800" height="549" border="0" /></p> | <center><p><img src="https://static.igem.org/mediawiki/2010/7/71/Ed10-SequentialBridge.JPG" width="800" height="549" border="0" /></p> | ||
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<p>At the first step of the process, the first antibiotic resistance gene and <i>sacB</i> are introduced alongside the first gene. The antibiotic resistance gene can then be replaced with the next gene and a second antibiotic resistance gene, thereby cycling the antibiotic resistance such that selection is different at each step. At the last step, both markers are removed and the final constructs can be selected for by growth on sucrose (growth on sucrose can also be used as a negative control at each stage, although this would only be to confirm the persistence of the marker).</p> | <p>At the first step of the process, the first antibiotic resistance gene and <i>sacB</i> are introduced alongside the first gene. The antibiotic resistance gene can then be replaced with the next gene and a second antibiotic resistance gene, thereby cycling the antibiotic resistance such that selection is different at each step. At the last step, both markers are removed and the final constructs can be selected for by growth on sucrose (growth on sucrose can also be used as a negative control at each stage, although this would only be to confirm the persistence of the marker).</p> | ||
| - | <p>The final construct would look as shown in Figure 2 | + | <p>The final construct would look as shown in <a href="https://static.igem.org/mediawiki/2010/a/a7/Ed10-FinalBridge.JPG">Figure 2</a>.</p><br> |
<center><p><img src="https://static.igem.org/mediawiki/2010/a/a7/Ed10-FinalBridge.JPG" width="800" height="199" border="0" /></p> | <center><p><img src="https://static.igem.org/mediawiki/2010/a/a7/Ed10-FinalBridge.JPG" width="800" height="199" border="0" /></p> | ||
<p><b>Figure 2:</b> The theoretical final construct after using BRIDGE to directly insert genes into the genome.</p><br><br></center> | <p><b>Figure 2:</b> The theoretical final construct after using BRIDGE to directly insert genes into the genome.</p><br><br></center> | ||
| - | <p> | + | <p>The steps above are purely theoretical and have not yet been tested, but the principle behind them is not too distant from the original method, so it would be nice to attempt it if anyone ever gets the chance.</p> |
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Revision as of 14:08, 27 October 2010
Future Work: Sequential addition
One of the future expansions of the BRIDGE protocol that we have discussed involves using it to directly introduce genes in the genome next to each other without using the BioBrick method before-hand. For example, if you wanted to insert four genes with the steps described in the protocol section, it would take eight steps. If you do this with the method shown in Figure 1 below, it would only take four steps.
Figure 1: Using BRIDGE to directly insert genes into the genome.
At the first step of the process, the first antibiotic resistance gene and sacB are introduced alongside the first gene. The antibiotic resistance gene can then be replaced with the next gene and a second antibiotic resistance gene, thereby cycling the antibiotic resistance such that selection is different at each step. At the last step, both markers are removed and the final constructs can be selected for by growth on sucrose (growth on sucrose can also be used as a negative control at each stage, although this would only be to confirm the persistence of the marker).
The final construct would look as shown in Figure 2.
Figure 2: The theoretical final construct after using BRIDGE to directly insert genes into the genome.
The steps above are purely theoretical and have not yet been tested, but the principle behind them is not too distant from the original method, so it would be nice to attempt it if anyone ever gets the chance.
Future Work: A working protocol
So far we have been unable
Future Applications
We want it to take over the world!
The advantages of the BRIDGE protocol over more traditional methods of BioBrick insertion have been documented here.
