Template:Leanna newesttoggle
From 2010.igem.org
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<tr><td><br>In the beginning, there was a UV Toggle | <tr><td><br>In the beginning, there was a UV Toggle | ||
(<a href="http://www.nature.com/nature/journal/v403/n6767/abs/403339a0.html">Collins, 2000</a>). <br> The 2010 MIT iGEM team saw that it was good, and decided to implement the Collins toggle in E.coli to create cells with bistable phenotypes. | (<a href="http://www.nature.com/nature/journal/v403/n6767/abs/403339a0.html">Collins, 2000</a>). <br> The 2010 MIT iGEM team saw that it was good, and decided to implement the Collins toggle in E.coli to create cells with bistable phenotypes. | ||
- | <div style="display: inline; background-color: #boc4de; width: | + | <div style="display: inline; background-color: #boc4de; width: 170px; float: right; font-size: x-small; border: 1px solid #b0c4de;"><a href="https://static.igem.org/mediawiki/2010/8/80/First.png" class="thickbox" title="The first patterned image created by exposing masked cells to UV light. The cells were made by co-transforming the Collins toggle plasmid pTSMa with our composite biobrick K415013."><img height=100px src="https://static.igem.org/mediawiki/2010/8/80/First.png"></a><br>The first picture of mCherry fluorescence induced by UV light.</div> |
The team planned for the toggle to control fluorescence and phage polymerization in response to exposing the cells to UV light. After much fine-tuning of the power of the UV exposure, the concentrations of AHL and IPTG, and mask cutting, a pattern of fluorescence finally emerged -- the first image.<br> | The team planned for the toggle to control fluorescence and phage polymerization in response to exposing the cells to UV light. After much fine-tuning of the power of the UV exposure, the concentrations of AHL and IPTG, and mask cutting, a pattern of fluorescence finally emerged -- the first image.<br> | ||
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But the team noticed a circle of cell death where the UV exposure had killed some of the cells in the lawn. They decided to make a pLPTa, a low power toggle that | But the team noticed a circle of cell death where the UV exposure had killed some of the cells in the lawn. They decided to make a pLPTa, a low power toggle that | ||
- | <div style="display: inline; background-color: #boc4de; width: 300px; float: right; font-size: small; border: 1px solid #b0c4de;"> | + | <div style="display: inline; background-color: #boc4de; width: 300px; float: right; font-size: x-small; border: 1px solid #b0c4de;"> |
<a href="https://static.igem.org/mediawiki/2010/e/eb/Igem_uv_prelim.png" class="thickbox" title="Power Modulation using the Low Power Toggle and cells that fluoresce red with UV induction."><img src="https://static.igem.org/mediawiki/2010/e/eb/Igem_uv_prelim.png" width=300px></a><br>Here we see cells controlled by the Low Power Toggle. The cells fluoresce red with UV induction, but at higher UV levels cell death can be seen in the green field.</div> still provided bistability, but required less UV power to induce a toggle switch. | <a href="https://static.igem.org/mediawiki/2010/e/eb/Igem_uv_prelim.png" class="thickbox" title="Power Modulation using the Low Power Toggle and cells that fluoresce red with UV induction."><img src="https://static.igem.org/mediawiki/2010/e/eb/Igem_uv_prelim.png" width=300px></a><br>Here we see cells controlled by the Low Power Toggle. The cells fluoresce red with UV induction, but at higher UV levels cell death can be seen in the green field.</div> still provided bistability, but required less UV power to induce a toggle switch. | ||
By site-directed mutagenesis, the team changed the lambda repressor (cI) gene in the Collins pTSMa to a cI that is more sensitive to cleavage by Rec-A, the enzyme activated by UV light exposure. | By site-directed mutagenesis, the team changed the lambda repressor (cI) gene in the Collins pTSMa to a cI that is more sensitive to cleavage by Rec-A, the enzyme activated by UV light exposure. |
Revision as of 03:40, 10 October 2010
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Overview Toggle Phage Mammalian Summary Acknowledgements
Materials & Methods Biosafety Journal Club
the bacterial uv toggle |
In the beginning, there was a UV Toggle (Collins, 2000). The 2010 MIT iGEM team saw that it was good, and decided to implement the Collins toggle in E.coli to create cells with bistable phenotypes. The team planned for the toggle to control fluorescence and phage polymerization in response to exposing the cells to UV light. After much fine-tuning of the power of the UV exposure, the concentrations of AHL and IPTG, and mask cutting, a pattern of fluorescence finally emerged -- the first image. Then the team decided to make the signal amplify itself, and record a movie to see its propogation. But the team noticed a circle of cell death where the UV exposure had killed some of the cells in the lawn. They decided to make a pLPTa, a low power toggle that still provided bistability, but required less UV power to induce a toggle switch. By site-directed mutagenesis, the team changed the lambda repressor (cI) gene in the Collins pTSMa to a cI that is more sensitive to cleavage by Rec-A, the enzyme activated by UV light exposure. |