Difference between revisions of "Team:Hong Kong UCCKE/Design"

 
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<h3 class="sectiontitle">Mechanism of the Promoter</h3>
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          <p style="text-align:left !important;">Our Project is divided into three sub-projects, the uric acid detector, the uric acid decomposer and the uric acid transporter, and they  are mentioned in the page of project description. These three BioBricks enclose the same promoter, and the mechanism of it will be explained below.</p>
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<h3 class="sectiontitle">Mechanism of the Uric Acid Concentration-Sensitive Promoter<sup><a href="#ref1b" id="ref1a">[1]</a></sup></h3>
 
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                 <p style="text-align:left !important;">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nunc vel varius libero, a mattis risus. Praesent sit amet ipsum ac odio venenatis consectetur. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nunc vel varius libero, a mattis risus. Praesent sit amet ipsum ac odio venenatis consectetur.</p>
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                 <p style="text-align:left !important;">The promoter is designed to be sensitive to the concentration of uric acid. This promoter controls the expression of the downstream gene and its session consists of a constitutive promoter J23100, a RBS B0034, a strong repressor KRAB-HucR and a double terminator B0015. HucR is itself a repressor. Its repressing ability is enhanced by KRAB. The resulting repressor is a chimeric mammalian urate-dependent transsilencer (mUTS).</p>
                <p style="text-align:left !important;">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nunc vel varius libero, a mattis risus. Praesent sit amet ipsum ac odio venenatis consectetur. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nunc vel varius libero, a mattis risus. Praesent sit amet ipsum ac odio venenatis consectetur.</p>
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<h3 class="sectiontitle" style="clear:both;">Lorem ipsum dolor sit amet last super long super long super long super long</h3>
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    <a href="https://static.igem.org/mediawiki/parts/1/13/T--Hong_Kong_UCCKE--Krab.jpg" title="Expression of a strong repressor (mUTS)">
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    <img src="https://static.igem.org/mediawiki/parts/1/13/T--Hong_Kong_UCCKE--Krab.jpg" alt="Expression of a strong repressor (mUTS)" style="width:100%; max-width:500px;">
                <p style="text-align:left !important;">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nunc vel varius libero, a mattis risus. Praesent sit amet ipsum ac odio venenatis consectetur. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nunc vel varius libero, a mattis risus. Praesent sit amet ipsum ac odio venenatis consectetur.</p>
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    </a><span class="imgcaption">Expression of a strong repressor (mUTS)</span></div>
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<br><p> HucO is an operative site for mUTS to bind into. When mUTS is binded to hucO, the expression of downstream gene is restricted according to the concentration of substrates. The presence of uric acid limits the binding of mUTS to hucO. The limitation varies as the concentration of uric acid.</p>
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<div class="pop">
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    <a href="https://static.igem.org/mediawiki/2017/a/a0/T--Hong_Kong_UCCKE--hucOgoi.jpg" title="Operator site and Gene of Interest">
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    <img src="https://static.igem.org/mediawiki/2017/a/a0/T--Hong_Kong_UCCKE--hucOgoi.jpg" alt="Operator site and Gene of Interest" style="width:100%; max-width:500px;">
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    </a><span class="imgcaption">Operator site and Gene of Interest</span></div>
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<p style="text-align:left !important;">mUTS binds to hucO and GFP is not expressed when uric acid is absent or at very low concentration. Alternatively, the complex detaches from hucO and GFP is expressed according to the concentration of uric acid.</p>
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<a href="https://static.igem.org/mediawiki/2017/3/3a/T--Hong_Kong_UCCKE--genenoua.jpg"  title="Expression without uric acid (-UA)">
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            <img src="https://static.igem.org/mediawiki/2017/3/3a/T--Hong_Kong_UCCKE--genenoua.jpg" alt="Expression without uric acid (-UA)" style="width:100%; max-width:500px;">
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<span class="imgcaption">Expression without uric acid (-UA)</span></div>
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<a href="https://static.igem.org/mediawiki/2017/3/37/T--Hong_Kong_UCCKE--genewifua.jpg" title="Expression with uric acid (+UA)">
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<img src="https://static.igem.org/mediawiki/2017/3/37/T--Hong_Kong_UCCKE--genewifua.jpg" alt="Expression with uric acid (+UA)" style="width:100%; max-width:500px;">
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<span class="imgcaption">Expression with uric acid (+UA)</span>
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<h3 class="sectiontitle" style="clear:both;" id="aim">Reference</h3>
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<div class="divider"></div>
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<p id="ref1b" class="ref-text-left"><a href="#ref1a">[1]</a>: Christian Kemmer, Marc Gitzinger, Marie Daoud-El Baba, Valentin Djonov, Jorg Stelling & Martin Fussenegger (2010) Self-sufficient control of urate homeostasis in mice by a synthetic circuit. Nature Biotechnology doi: 10.1038/nbt.1617 </p>
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Latest revision as of 09:22, 1 November 2017

Our Project is divided into three sub-projects, the uric acid detector, the uric acid decomposer and the uric acid transporter, and they are mentioned in the page of project description. These three BioBricks enclose the same promoter, and the mechanism of it will be explained below.

Mechanism of the Uric Acid Concentration-Sensitive Promoter[1]

The promoter is designed to be sensitive to the concentration of uric acid. This promoter controls the expression of the downstream gene and its session consists of a constitutive promoter J23100, a RBS B0034, a strong repressor KRAB-HucR and a double terminator B0015. HucR is itself a repressor. Its repressing ability is enhanced by KRAB. The resulting repressor is a chimeric mammalian urate-dependent transsilencer (mUTS).

Expression of a strong repressor (mUTS) Expression of a strong repressor (mUTS)

HucO is an operative site for mUTS to bind into. When mUTS is binded to hucO, the expression of downstream gene is restricted according to the concentration of substrates. The presence of uric acid limits the binding of mUTS to hucO. The limitation varies as the concentration of uric acid.

Operator site and Gene of Interest Operator site and Gene of Interest

mUTS binds to hucO and GFP is not expressed when uric acid is absent or at very low concentration. Alternatively, the complex detaches from hucO and GFP is expressed according to the concentration of uric acid.

Expression without uric acid (-UA) Expression without uric acid (-UA)
Expression with uric acid (+UA) Expression with uric acid (+UA)

Reference

[1]: Christian Kemmer, Marc Gitzinger, Marie Daoud-El Baba, Valentin Djonov, Jorg Stelling & Martin Fussenegger (2010) Self-sufficient control of urate homeostasis in mice by a synthetic circuit. Nature Biotechnology doi: 10.1038/nbt.1617