Difference between revisions of "Team:Jilin China/Result/Version 1"

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   <p><span>TOOLKITS</span><br>VERSION 1.0</p>
 
   <p><span>TOOLKITS</span><br>VERSION 1.0</p>
 
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    <li class="full-width"><div><h2>余戎镇 正在添加底部内容</h2></div></li>
 
 
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<p>SynRT toolkits version 1.0 is the first generation thermosensors collection we provide to users. It contains 48 heat-induced RNA-based thermosensors. </p>
 
<p>SynRT toolkits version 1.0 is the first generation thermosensors collection we provide to users. It contains 48 heat-induced RNA-based thermosensors. </p>
 
<p>We constructed in a total of 150 heat-inducible RNA thermosensors using goldengate assembly described in the <a href="https://2018.igem.org/Team:Jilin_China/Construction">Construction page</a>. And the measurement method was describled in <a href="https://2018.igem.org/Team:Jilin_China/Construction">Measurement page</a>. </p>
 
<p>We constructed in a total of 150 heat-inducible RNA thermosensors using goldengate assembly described in the <a href="https://2018.igem.org/Team:Jilin_China/Construction">Construction page</a>. And the measurement method was describled in <a href="https://2018.igem.org/Team:Jilin_China/Construction">Measurement page</a>. </p>
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<p>We measured the activities of these thermosensors at six different temperatures: 29, 31, 35, 37, 39 and 42℃.Upon screening, we selected 48 heat-inducible RNA thermosensors out of the 150 members. And figure 1 shows the 48 thermosensors' activity. The positive control we designed won't form stem-loop structure with SD sequence by software prediction, and the experimental charaterization is described in Measurement page. The normalized fluorescence are used to describe the change of activity compared with positive control. We find that all of these thermosensors' expression level increase at elevated temperature, and the extent of increase relative to temperature is different. </p>
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<li><b>Activities of thermosensors increase at elevated temperature</b></li>
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<p>We measured the activities of these thermosensors at six different temperatures: 29, 31, 35, 37, 39 and 42℃, and then fitted curve. Through the curve fitting, we discarded some thermosensors, which cannot fit the curve or have too high or too low melting temperature. Upon screening, we selected 48 heat-inducible RNA thermosensors out of the 150 members. And <b>figure 1</b> shows the 48 thermosensors' activity. The positive control we designed won't form stem-loop structure with SD sequence by software prediction, and the experimental charaterization is described in <a href="https://2018.igem.org/Team:Jilin_China/Measurement">Measurement page</a>. The normalized fluorescence are used to describe the change of activity compared with positive control. We find that all of these thermosensors' expression level increase at elevated temperature, and the extent of increase relative to temperature is different. </p>
 
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<img src="https://static.igem.org/mediawiki/2018/8/82/T--Jilin_China--result--RTbar.png" width="95%" length="95%"></img>
 
<img src="https://static.igem.org/mediawiki/2018/8/82/T--Jilin_China--result--RTbar.png" width="95%" length="95%"></img>
 
<p>To quantify different features of this library, we computed the fold-change of the response in the given temperature range. As the figure shows, from 29 to 37℃, the fold change ranges from x fold to x fold. And from 37 to 42℃, the fold change ranges from x fold to x fold. These fold-changes were higher than positive control.  These results show that there is a diverse set of heat-inducible RNA thermosensors in toolkit.</p>
 
<p>To quantify different features of this library, we computed the fold-change of the response in the given temperature range. As the figure shows, from 29 to 37℃, the fold change ranges from x fold to x fold. And from 37 to 42℃, the fold change ranges from x fold to x fold. These fold-changes were higher than positive control.  These results show that there is a diverse set of heat-inducible RNA thermosensors in toolkit.</p>
 
<img src="https://static.igem.org/mediawiki/2018/6/62/T--Jilin_China--result--RTfold.png" width="95%" length="95%"></img>
 
<img src="https://static.igem.org/mediawiki/2018/6/62/T--Jilin_China--result--RTfold.png" width="95%" length="95%"></img>
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<p>Discussion: as these results show, the heat-inducible RNA thermosensor we designed can work. The fluorescence value increases with temperature elevated. Besides, the difference in fluorescence intensity and the rate of increase points to the diversity in thermosensor response. We think the sequence change in stem length, loop size, and mismatched or bulges in the stem give rise to the diversity in thermosensor response.</p>
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<p><b>Conclusion:</b> as these results show, the heat-inducible RNA thermosensor we designed can work. The fluorescence value increases with temperature elevated. Besides, the difference in fluorescence intensity and the rate of increase points to the diversity in thermosensor response. We think the sequence change in stem length, loop size, and mismatched or bulges in the stem give rise to the diversity in thermosensor response.</p>
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  <li><b>Thermosensors' features can be computed through fitted curve</b></li>
 
<p>Combined with our human practice work we have mentioned in the Overview page. We also have built switching behavior fitting model to describe the continuous switching behavior of thermosenors.</p>
 
<p>Combined with our human practice work we have mentioned in the Overview page. We also have built switching behavior fitting model to describe the continuous switching behavior of thermosenors.</p>
 
<center><a href="https://2018.igem.org/Team:Jilin_China/Model">>For more details about our model, you can visit Model page.<</a></center>
 
<center><a href="https://2018.igem.org/Team:Jilin_China/Model">>For more details about our model, you can visit Model page.<</a></center>
 
<p>From the fitted curve, we can get the melting temperature, intensity and sensitivity of thermosensors. Based on these datas, we classified thermosensors, and develop a search engine -- SynRT Explorer. You can visit our <a href="https://2018.igem.org/Team:Jilin_China/Part/Search_Engine">Search Engine page</a> to use it. </p>
 
<p>From the fitted curve, we can get the melting temperature, intensity and sensitivity of thermosensors. Based on these datas, we classified thermosensors, and develop a search engine -- SynRT Explorer. You can visit our <a href="https://2018.igem.org/Team:Jilin_China/Part/Search_Engine">Search Engine page</a> to use it. </p>
 
<p>You can see the individual result as followed:</p>
 
<p>You can see the individual result as followed:</p>
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Revision as of 21:54, 14 October 2018

TOOLKITS
VERSION 1.0


VERSION 1.0

  • Results

    SynRT toolkits version 1.0 is the first generation thermosensors collection we provide to users. It contains 48 heat-induced RNA-based thermosensors.

    We constructed in a total of 150 heat-inducible RNA thermosensors using goldengate assembly described in the Construction page. And the measurement method was describled in Measurement page.

  • Activities of thermosensors increase at elevated temperature

    • We measured the activities of these thermosensors at six different temperatures: 29, 31, 35, 37, 39 and 42℃, and then fitted curve. Through the curve fitting, we discarded some thermosensors, which cannot fit the curve or have too high or too low melting temperature. Upon screening, we selected 48 heat-inducible RNA thermosensors out of the 150 members. And figure 1 shows the 48 thermosensors' activity. The positive control we designed won't form stem-loop structure with SD sequence by software prediction, and the experimental charaterization is described in Measurement page. The normalized fluorescence are used to describe the change of activity compared with positive control. We find that all of these thermosensors' expression level increase at elevated temperature, and the extent of increase relative to temperature is different.


    To quantify different features of this library, we computed the fold-change of the response in the given temperature range. As the figure shows, from 29 to 37℃, the fold change ranges from x fold to x fold. And from 37 to 42℃, the fold change ranges from x fold to x fold. These fold-changes were higher than positive control. These results show that there is a diverse set of heat-inducible RNA thermosensors in toolkit.

    Conclusion: as these results show, the heat-inducible RNA thermosensor we designed can work. The fluorescence value increases with temperature elevated. Besides, the difference in fluorescence intensity and the rate of increase points to the diversity in thermosensor response. We think the sequence change in stem length, loop size, and mismatched or bulges in the stem give rise to the diversity in thermosensor response.

  • Thermosensors' features can be computed through fitted curve

    • Combined with our human practice work we have mentioned in the Overview page. We also have built switching behavior fitting model to describe the continuous switching behavior of thermosenors.

    >For more details about our model, you can visit Model page.<

    From the fitted curve, we can get the melting temperature, intensity and sensitivity of thermosensors. Based on these datas, we classified thermosensors, and develop a search engine -- SynRT Explorer. You can visit our Search Engine page to use it.

    You can see the individual result as followed: