Difference between revisions of "Team:NEFU China/Suicide"

 
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<meta charset="utf-8">
 
<meta charset="utf-8">
  
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<title>Suicide</title>
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<title>Information Destruction</title>
  
 
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<ul class="firstmenu" style="float: left">
 
<ul class="firstmenu" style="float: left">
 
 
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<ul id="sub_02">
 
<ul id="sub_02">
 
<li><a href="https://2018.igem.org/Team:NEFU_China/Background" target="_self">BACKGROUND</a></li>
 
<li><a href="https://2018.igem.org/Team:NEFU_China/Background" target="_self">BACKGROUND</a></li>
−
<li><a href="https://2018.igem.org/Team:NEFU_China/Description" target="_self">DESCRIPTION</a></li>
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<li><a href="https://2018.igem.org/Team:NEFU_China/Description" target="_self">DESCRIPTION &amp; DESIGN</a></li>
−
<li><a href="https://2018.igem.org/Team:NEFU_China/Design" target="_self">DESIGN</a></li>
+
<li><a href="https://2018.igem.org/Team:NEFU_China/Coding book" target="_self">CODE BOOK</a></li>
−
<li><a href="https://2018.igem.org/Team:NEFU_China/Coding book" target="_self">CODING BOOK</a></li>
+
 
  </ul>
 
  </ul>
 
  </li>
 
  </li>
−
  <li class="mainlevel" id="mainlevel_03">
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<li class="mainlevel" id="mainlevel_03">
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  <a href="https://2018.igem.org/Team:NEFU_China/Basic parts"><img id="parts" src="https://static.igem.org/mediawiki/2018/5/58/T--NEFU_China--_PARTS.png">PARTS</a>
+
  <a href="https://2018.igem.org/Team:NEFU_China/Demonstrate"><img id="parts" src="https://static.igem.org/mediawiki/2018/6/62/T--NEFU_China--_RESULTS.png">EXPERIMENTS</a>
 
  <ul id="sub_03">
 
  <ul id="sub_03">
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Basic parts" target="_self">BASIC PARTS</a></li>
+
  <li><a href="https://2018.igem.org/Team:NEFU_China/Lock_Key" target="_self">LOCK &amp; KEY</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Composite parts" target="_self">COMPOSITE PARTS</a></li>
+
  <li><a href="https://2018.igem.org/Team:NEFU_China/Suicide" target="_self">INFORMATION DESTRUCTION</a></li>
 +
  <li><a href="https://2018.igem.org/Team:NEFU_China/Splicing" target="_self">Pre-RNA SPLICING</a></li>
 +
  <li><a href="https://2018.igem.org/Team:NEFU_China/Demonstrate" target="_self">DEMONSTRATE</a></li>
 +
  <hr>
 +
  <li><a href="https://2018.igem.org/Team:NEFU_China/Basic_Part" target="_self">BASIC PARTS</a></li>
 +
  <li><a href="https://2018.igem.org/Team:NEFU_China/Composite_Part" target="_self">COMPOSITE PARTS</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Improve" target="_self">IMPROVEMENT PARTS</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Improve" target="_self">IMPROVEMENT PARTS</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Parts collection" target="_self">PARTS COLLECTION</a></li>
+
  <li><a href="https://2018.igem.org/Team:NEFU_China/Part_Collection" target="_self">PARTS COLLECTION</a></li>
−
  </ul>
+
  <hr>
−
  </li>
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Notebook" target="_self">NOTEBOOK</a></li>
−
      <li class="mainlevel" id="mainlevel_04">
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Protocol" target="_self">PROTOCOL</a></li>
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  <a href="https://2018.igem.org/Team:NEFU_China/Lock_Key"><img id="results" src="https://static.igem.org/mediawiki/2018/6/62/T--NEFU_China--_RESULTS.png">RESULTS</a>
+
−
  <ul id="sub_04">
+
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Lock_Key" target="_self">LOCK &amp; KEY</a></li>
+
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Suicide" target="_self">SUICIDE</a></li>
+
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Splicing" target="_self">SPLICING</a></li>
+
 
  </ul>
 
  </ul>
 
  </li>
 
  </li>
 +
 
 
  <li class="mainlevel" id="mainlevel_05">
 
  <li class="mainlevel" id="mainlevel_05">
 
  <a href="https://2018.igem.org/Team:NEFU_China/Model"><img id="model" src="https://static.igem.org/mediawiki/2018/0/0c/T--NEFU_China--_MODEL.png">MODEL</a>
 
  <a href="https://2018.igem.org/Team:NEFU_China/Model"><img id="model" src="https://static.igem.org/mediawiki/2018/0/0c/T--NEFU_China--_MODEL.png">MODEL</a>
 
  <ul id="sub_05">
 
  <ul id="sub_05">
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Model" target="_self">OVERVIEW</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Model" target="_self">OVERVIEW</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Model1" target="_self">MODEL1</a></li>
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Model1" target="_self">CORRESPONDING COEFFICIENT</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Model2" target="_self">MODEL2</a></li>
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Model2" target="_self">KILLING MODEL</a></li>
 
  </ul>
 
  </ul>
 
  </li>
 
  </li>
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  <ul id="sub_06">
 
  <ul id="sub_06">
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Software" target="_self">OVERVIEW</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Software" target="_self">OVERVIEW</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Software1" target="_self">SOFTWARE1</a></li>
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Software1" target="_self">CODING</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Software2" target="_self">SOFTWARE2</a></li>
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Software2" target="_self">MISLEADING</a></li>
 +
  <li><a href="https://2018.igem.org/Team:NEFU_China/Software3" target="_self">WORDSEGMENT</a></li>
 
  </ul>
 
  </ul>
 
  </li>
 
  </li>
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  <li><a href="https://2018.igem.org/Team:NEFU_China/Attributions" target="_self">ATTRIBUTIONS</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Attributions" target="_self">ATTRIBUTIONS</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Members" target="_self">MEMBERS</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Members" target="_self">MEMBERS</a></li>
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Sponsoring" target="_self">SPONSORING</a></li>
 
 
  </ul>
 
  </ul>
 
  </li>
 
  </li>
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  <a href="https://2018.igem.org/Team:NEFU_China/Human_Practices"><img id="humanpractice" src="https://static.igem.org/mediawiki/2018/9/91/T--NEFU_China--_HUMANPRACTICE.png">HUMAN PRACTICE</a>
 
  <a href="https://2018.igem.org/Team:NEFU_China/Human_Practices"><img id="humanpractice" src="https://static.igem.org/mediawiki/2018/9/91/T--NEFU_China--_HUMANPRACTICE.png">HUMAN PRACTICE</a>
 
  <ul id="sub_08">
 
  <ul id="sub_08">
 +
  <li><a href="https://2018.igem.org/Team:NEFU_China/Human_Practices" target="_self">OVERVIEW</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Gold_integrated" target="_self">GOLD INTEGRATED</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Gold_integrated" target="_self">GOLD INTEGRATED</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Silver" target="_self">SILVER</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Silver" target="_self">SILVER</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Collaborations" target="_self">COLLABORTION</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Collaborations" target="_self">COLLABORTION</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Public_Engagement" target="_self">EDUCATION &amp; PUBLIC ENGAGEMENT</a></li>
 
  <li><a href="https://2018.igem.org/Team:NEFU_China/Public_Engagement" target="_self">EDUCATION &amp; PUBLIC ENGAGEMENT</a></li>
−
  </ul>
 
−
  </li>
 
−
  <li class="mainlevel" id="mainlevel_09">
 
−
  <a href="https://2018.igem.org/Team:NEFU_China/Notebook"><img id="notebook" src="https://static.igem.org/mediawiki/2018/c/cb/T--NEFU_China--_NOTEBOOK.png">NOTEBOOK</a>
 
−
  <ul id="sub_09">
 
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Notebook" target="_self">OVERVIRW</a></li>
 
−
  <li><a href="https://2018.igem.org/Team:NEFU_China/Protocol" target="_self">PROTOCOL</a></li>
 
 
  </ul>
 
  </ul>
 
  </li>
 
  </li>
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</div>
 
</div>
 
<div id="banner">  
 
<div id="banner">  
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<img src="https://static.igem.org/mediawiki/2018/0/05/T--NEFU_China--DNA.png" alt="banner" id="banner-img">
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<img src="https://static.igem.org/mediawiki/2018/2/26/T--NEFU_China--result10.png" alt="banner" id="banner-img">
 
</div>
 
</div>
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<div id="background-content">
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<div class="layer-bottom">
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<h1>Suicide</h1>
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    <p>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.<br>
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        But since few have marked down their own prices in line with the metal's fall, they will be able to recoup much of the difference. Not so the producers, whose income is directly related to the fluctuating daily price on the London Metal Exchange.<br>
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        The Japanese have their electronics, the Germans their engineering. But when it comes to command of global markets, the U.S. owns the service sector.<br>
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        Meanwhile, pressure has been growing from the car companies. GM ships about 60% of its cars and trucks with Ryder, while Chrysler ships some 40%.<br>
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        First of all, current modest demand growth will not support any more increases that large. Second, now that manufacturers have worked to get their inventories lower, they will be cautious about adding goods in coming months.<br>
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These are two steps:
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</p>
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<ol>
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<li>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.
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</li>
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<li>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.
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</li>
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</ol>
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<p>
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The Japanese have their electronics, the Germans their engineering. But when it comes to command of global markets, the U.S. owns the service sector.<br>
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</p>
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<img src="https://static.igem.org/mediawiki/2018/5/58/T--NEFU_China--Figure_1.png" alt="Figure1" id="Figure1-img">
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<h2 id="Figure1-title">Figure 1: This is Figure 1.</h2>
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<p>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.<br>
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<h1>Suicide first title</h1>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.<br>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.<br>
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So far, the national trends in costs for wages, salaries, and benefits have glossed over these concerns. The growth in labor costs continued to slow in the second quarter - a pattern that held true in all major regions. However, the slowdown in labor costs is due solely to sharp cutbacks in what companies, mainly large corporations, are paying for benefits, which make up about a fourth of total compensation costs nationally. Because of slower growth in health care costs, workers' compensation, and state unemployment insurance, benefits grew only 2.6% during the past year, the lowest pace on record.<br>
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<h1 style="font-size: 65px;color: orange!important;height: 110px;"><br>
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Information Destruction</h1>
 
<p>
 
<p>
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The Japanese have their electronics, the Germans their engineering. But when it comes to command of global markets, the U.S. owns the service sector.<br>
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</p>
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    <strong><span style="
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<br><br><br>
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    color: orange;
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    font-size: 26px;
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">(α factor induced apoptosis)</span></strong><br>
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 +
    <br>
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<span style="
 +
    font-size: 45px;
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    color: orange;
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">Yeast: a-type yeast.</span><br><br>
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<strong><span>Function:</span></strong> The expression of the Fig2C promoter can be induced by adding a mating factor. We want to use this promoter to express the Bax(alpha) gene. We used the enhanced green fluorescent protein (EGFP) to test the effect of the Fig2C promoter. When the Fig2C promoter is induced, EGFP is expressed. In this way, we can detect the strengthen of the Fig2C promoter using EGFP as a reporter.<br><br>
 +
<strong>Vector construction:</strong>
 +
We first inserted the Fig2C promoter and EGFP coding sequence into the pesc-ura plasmid. And the constructed plasmid is shown in Figure 1.
 +
<br><br>
 +
</p><div align="center">
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<img src="https://static.igem.org/mediawiki/2018/c/cc/T--NEFU_China--inf01.png" style="width:60%;"><br>
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 +
<span style="
 +
    font-size: 24px;
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">Figure 1: A: pFig2C-EGFP  B: pFig2C-Bax(alpha)</span>
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<br>
 
</div>
 
</div>
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<div id="background-reference">
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<br>
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<h1>Reference</h1>
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<p> <a href="#"> [1] Pu, Jinyue and Zinkus-Boltz, Julia and Dickinson, Bryan C. (2017) Evolution of a split RNA polymerase as a versatile biosensor platform. <em><em>Nat Chem Biol 13</em></em> , 432-438 </a>
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<p><strong style="
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    font-size: 26px;
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">Functional verification: </strong>
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    <span style="
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    /* text-align: justify; */
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">
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In order to verify whether α factor induces the expression of the Fig2C promoter, we transferred the constructed plasmids into yeast, and induced them with α factor. We observed the fluorescence in the transformed yeast cells under a fluorescence microscope (Figure 2). In addition, we also did quantitative PCR for EGFP mRNA. The results showed that EGFP expression significantly increased at 12 h (Figure 3), compared with the control group. These results showed that α factor can induce expression of the Fig2C promoter.
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</span>
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</p><br>
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<br>
 
<br>
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<a href="#"> [2] Pu, Jinyue and Zinkus-Boltz, Julia and Dickinson, Bryan C. (2017) Evolution of a split RNA polymerase as a versatile biosensor platform. <em><em>Nat Chem Biol 13</em></em> , 432-438 </a>
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<div align="center">
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<br>
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    <img src="https://static.igem.org/mediawiki/2018/e/eb/T--NEFU_China--inf02.png" style="width:60%;"><br>
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<a href="#"> [3] Pu, Jinyue and Zinkus-Boltz, Julia and Dickinson, Bryan C. (2017) Evolution of a split RNA polymerase as a versatile biosensor platform. <em><em>Nat Chem Biol 13</em></em> , 432-438 </a>
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<span style="
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<br>
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    font-size: 24px;
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<a href="#"> [4] Pu, Jinyue and Zinkus-Boltz, Julia and Dickinson, Bryan C. (2017) Evolution of a split RNA polymerase as a versatile biosensor platform. <em><em>Nat Chem Biol 13</em></em> , 432-438 </a>
+
">Figure 2: Fluorescence image of transformed yeast cells at 12h time <br>point after cultivation with (A) and without (B) 0.4 g of α factor dry powder.</span>
−
<br>
+
    <br>
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<a href="#"> [5] Pu, Jinyue and Zinkus-Boltz, Julia and Dickinson, Bryan C. (2017) Evolution of a split RNA polymerase as a versatile biosensor platform. <em><em>Nat Chem Biol 13</em></em> , 432-438 </a>
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        <br>
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<br>
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    <img src="https://static.igem.org/mediawiki/2018/6/67/T--NEFU_China--inf03.png" style="width:60%;"><br>
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<a href="#"> [6] Pu, Jinyue and Zinkus-Boltz, Julia and Dickinson, Bryan C. (2017) Evolution of a split RNA polymerase as a versatile biosensor platform. <em><em>Nat Chem Biol 13</em></em> , 432-438 </a>s
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    <span style="
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</p>
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    font-size: 24px;
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<br><br><br>
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">Figure 3: Quantitative PCR for EGFP mRNA from Yeast with or without α factor.</span><br>
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    <strong>Functional verification of Bax(alpha) gene</strong><br>
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<td>
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We transferred the constructed plasmid pFig2C-EGFP into yeast, we call it “Spy Yeast”, and induced them with α factor. And we define yeast that secretes alpha factor as “Killer yeast”. 10 ml of the spy yeast culture solution and 50ul of the killer yeast culture solution(OD600nm is about 1.4) were mixed for cocultivation.<br>
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<h1>Userfull links</h1>
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Meanwhile, we used the cocultured spy yeast without integrated Bax gene and the killer yeast as a control group. Finally, we could determine that the spy yeast could be completely eliminated after 14 hours of the cocultivation (Figure 4).<br>
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<h2>
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    </p>
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<a href="https://2018.igem.org/Team:NEFU_China/NEFU_China">Home</a>
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    <br>
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&nbsp;&nbsp;&nbsp;&nbsp;
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<div align="center">
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<a href="https://2018.igem.org/Team:NEFU_China/MODEL/Overview">Model</a>
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<img src="https://static.igem.org/mediawiki/2018/9/9f/T--NEFU_China--inf04.png" style="width:60%;"><br>
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&nbsp;&nbsp;&nbsp;
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<a href="https://2018.igem.org/Team:NEFU_China/PROJECT/Description">Project</a>
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    <span style="
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    &nbsp;&nbsp;
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    font-size: 24px;
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    <a href="https://2018.igem.org/Team:NEFU_China/SOFTWARE/Overview">Software</a>
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">Figure 4: OD(600nm) value of two experiment groups.</span>
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</h2>
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    <br>
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</div>
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<h2>
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<a href="https://2018.igem.org/Team:NEFU_China/PARTS/Basic parts">Parts</a>
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<p></p>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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<a href="https://2018.igem.org/Team:NEFU_China/TEAM/Members">Teams</a>
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&nbsp;&nbsp;
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<a href="https://2018.igem.org/Team:NEFU_China/RESULTS/Leaders">Results</a>
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&nbsp;
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<a href="https://2018.igem.org/Team:NEFU_China/NOTEBOOK/Overview">Notebook</a>
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</h2>
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<h2>
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<a href="https://2018.igem.org/Team:NEFU_China/HUMAN PRACTICE/Gold_integrated">Human &nbsp;Practice</a>
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</h2>
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</td>
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<td style="padding-left: 70px!important;">
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<h1>Follow us</h1>
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<img alt="facebook" src="https://static.igem.org/mediawiki/2018/b/b5/T--NEFU_China--facebook.png">
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<img alt="wehchat" src="https://static.igem.org/mediawiki/2018/c/ca/T--NEFU_China--wechat.png">
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<img alt="wehchat" src="https://static.igem.org/mediawiki/2018/c/ca/T--NEFU_China--wechat.png">
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</td>
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<td style="padding-left: 70px!important;">
+
−
<h1>Contact us</h1>
+
−
<h2>iGEM-NEFU_China2018</h2>
+
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<h2>Email: hexinglu@nefu.edu.cn</h2>
+
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<h3>No.26 Hexing Road, Xiangfang <br>District, Harbin, Heilongjiang <br>Province 150000</h3>
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Latest revision as of 20:58, 17 October 2018

Information Destruction


Information Destruction

(α factor induced apoptosis)

Yeast: a-type yeast.

Function: The expression of the Fig2C promoter can be induced by adding a mating factor. We want to use this promoter to express the Bax(alpha) gene. We used the enhanced green fluorescent protein (EGFP) to test the effect of the Fig2C promoter. When the Fig2C promoter is induced, EGFP is expressed. In this way, we can detect the strengthen of the Fig2C promoter using EGFP as a reporter.

Vector construction: We first inserted the Fig2C promoter and EGFP coding sequence into the pesc-ura plasmid. And the constructed plasmid is shown in Figure 1.


Figure 1: A: pFig2C-EGFP B: pFig2C-Bax(alpha)

Functional verification: In order to verify whether α factor induces the expression of the Fig2C promoter, we transferred the constructed plasmids into yeast, and induced them with α factor. We observed the fluorescence in the transformed yeast cells under a fluorescence microscope (Figure 2). In addition, we also did quantitative PCR for EGFP mRNA. The results showed that EGFP expression significantly increased at 12 h (Figure 3), compared with the control group. These results showed that α factor can induce expression of the Fig2C promoter.




Figure 2: Fluorescence image of transformed yeast cells at 12h time
point after cultivation with (A) and without (B) 0.4 g of α factor dry powder.


Figure 3: Quantitative PCR for EGFP mRNA from Yeast with or without α factor.

Functional verification of Bax(alpha) gene
We transferred the constructed plasmid pFig2C-EGFP into yeast, we call it “Spy Yeast”, and induced them with α factor. And we define yeast that secretes alpha factor as “Killer yeast”. 10 ml of the spy yeast culture solution and 50ul of the killer yeast culture solution(OD600nm is about 1.4) were mixed for cocultivation.
Meanwhile, we used the cocultured spy yeast without integrated Bax gene and the killer yeast as a control group. Finally, we could determine that the spy yeast could be completely eliminated after 14 hours of the cocultivation (Figure 4).



Figure 4: OD(600nm) value of two experiment groups.