Team:NYMU-Taipei/ymiq2.html

From 2012.igem.org

(Difference between revisions)
 
(26 intermediate revisions not shown)
Line 80: Line 80:
<div class="title">Methods</div>
<div class="title">Methods</div>
<div align="left">
<div align="left">
-
   <p><span class="subtitle">Resistance of Synechococcus SP. PCC 7002 to 3 - (3,4-dichlorophenyl) - 1,1 – dimethylurea (DCMU)</span></p>
+
   <p><span class="subtitle">Resistance of Cyanobacteria (Synechococcus SP. PCC 7002) to Sulfide compound</span></p>
</div>
</div>
<div align="left">
<div align="left">
-
   <p>From the previous research, we discovered that the concentration of 3 - (3,4 - dichlorophenyl) - 1,1 – dimethylurea (DCMU) must be adjusted to meet our requirement. Under certain DCMU concentration, the presence of sulfide would be extreme decisive condition which determines whether the colonies live or die. In this experiment, DCMU is diluted with A2 medium to explore the relationship between DCMU concentration and cell growth. Sodium sulfide is added to the experimental group and its initial concentration is controlled to 10 mM. <br />
+
   <p>Several Cyanobacteria have Sulfide-Quinone Reductase (sqr) and thus the ability to deprive electron from sulfide compound. According to both databases of NCBI and KEGG, the sqr in Synechococcus SP. PCC 7002 shared great similarity with that of Oscillatoria limnetica, which is reported to exhibit anoxygenic photosynthesis by consumed sulfide anion. Since we planned to express sqr from Synechococcus SP. PCC 7002 in Synechococcus SP. PCC 7942 and Escherichia coli, the experiment was designed to testify the property of the sqr. DCMU was added in the medium to inhibit photosystem II, and therefore only sodium sulfide in the medium can provide electron for carbon photoassimilation. By creating different dilution of sodium sulfide, we expected that the more sodium sulfide was present, the better the cell grew.   <br />
    
    
    
    
-
   <div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/c/cb/Ymiq3.gif" width="538" height="190" border="0" align="center"  /><br />
+
   <div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/5/5e/Ymiq3.png" width="469" height="276" border="0" align="center"  /><br />
     DCMU structure and its mechanism on photosynthesis<br />
     DCMU structure and its mechanism on photosynthesis<br />
     http://en.wikipedia.org/wiki/File:Diuron.png <br />
     http://en.wikipedia.org/wiki/File:Diuron.png <br />
Line 96: Line 96:
</div>
</div>
     <p>From the previous  studies, it is suggested that <em>Synechococcus  SP. PCC 7002 </em>is able to metabolize sulfide compounds. We took advantage of  the results in our last experiment and adjusted the concentration of DCMU to an  appropriate degree. Since sulfide would become the main reducing energy for photoassimilation  under the effect of DCMU, we believe the more sulfide concentration in the  wells, the better cell growth would be observed.</p>
     <p>From the previous  studies, it is suggested that <em>Synechococcus  SP. PCC 7002 </em>is able to metabolize sulfide compounds. We took advantage of  the results in our last experiment and adjusted the concentration of DCMU to an  appropriate degree. Since sulfide would become the main reducing energy for photoassimilation  under the effect of DCMU, we believe the more sulfide concentration in the  wells, the better cell growth would be observed.</p>
-
       <div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/c/c3/Ymiq4.png" width="573" height="278" border="0" align="center"  /></span><br />
+
       <div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/2/23/Ymiq4-1.png" width="557" height="415" border="0" align="center"  /></span><br />
 +
<br />
 +
 
 +
 +
<p><span class="subtitle">The effect of sodium sulfide on Synechococcus SP. PCC 7942 growth rate</span></p>
 +
</div>
 +
 
 +
    <p>After thoroughly examined the ability of sqr in Synechococcus SP. PCC 7002, we planned to conduct a series of similar experiments on Synechococcus SP. PCC 7942. Except for the cultivation medium, other growing conditions remained the same. Instinctively, the strain expressing sqr should grow better than the wile type strain. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper.</p>
 +
      <div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/5/57/Ymiq4-2.png" width="573" height="299" border="0" align="center"  /></span><br />
 +
  <div align="left">
 +
      <p><span class="subtitle">Sulfide concentration and the growth of sqr expressing strain Synechococcus SP. PCC 7942</span></p>
 +
</div>
 +
  <div align="left">
 +
    <p>It was expected that SQR expressing strain and wild type counterpart would have different growth rate under the presence of sulfide compounds. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper. As the result, we analyze H2S amount to detect whether sqr gene work or not. Therefore, we perform Chemical microvolume turbidimetry method to detect H2S concentration (see Sulfur Oxide Terminator part)</div><br />
 +
 
 +
<div align="left">
 +
      <p><span class="subtitle">Sulfide oxidation in Escherichia coli expressing sulfide-quinone reductase gene</span></p>
 +
</div>
 +
  <div align="left">
 +
    <p>Repots have it that Escherichia coli can express functional sulfide-quinone reductase (SQR). Therefore, we slightly adjusted the previous experiment and applied to the SQR gene from Synechococcus SP. PCC 7002. With methylene blue method, we would test the efficiency of SQR sulfide oxidation. Since such method involved in measurement of optical density, it is more appropriate to perform such experiment on colorless bacteria instead of engineered cyanobacteria strain.</div><br />
 +
      <br />
 +
 
   </div>
   </div>
</div>
</div>
Line 114: Line 135:
                 <li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li>
                 <li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li>
                 <li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li>
                 <li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li>
-
                 <li><a title="Measurements" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Measurements</a></li>
+
                 <li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li>
-
                 <li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li>
+
                 <li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li>
             </ul>
             </ul>
         </li>
         </li>
Line 130: Line 151:
                 Industrial Waste Detection</a></li>
                 Industrial Waste Detection</a></li>
                 <li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li>
                 <li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li>
-
                 <li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li>
+
                 <li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymis8.html">Further Experiments after Asia Jamboree</a></li>
             </ul>
             </ul>
         </li>
         </li>

Latest revision as of 01:48, 27 October 2012

NYMU iGEM

Methods

Resistance of Cyanobacteria (Synechococcus SP. PCC 7002) to Sulfide compound

Several Cyanobacteria have Sulfide-Quinone Reductase (sqr) and thus the ability to deprive electron from sulfide compound. According to both databases of NCBI and KEGG, the sqr in Synechococcus SP. PCC 7002 shared great similarity with that of Oscillatoria limnetica, which is reported to exhibit anoxygenic photosynthesis by consumed sulfide anion. Since we planned to express sqr from Synechococcus SP. PCC 7002 in Synechococcus SP. PCC 7942 and Escherichia coli, the experiment was designed to testify the property of the sqr. DCMU was added in the medium to inhibit photosystem II, and therefore only sodium sulfide in the medium can provide electron for carbon photoassimilation. By creating different dilution of sodium sulfide, we expected that the more sodium sulfide was present, the better the cell grew.


DCMU structure and its mechanism on photosynthesis
http://en.wikipedia.org/wiki/File:Diuron.png

Sodium sulfide concentration and cell growth

From the previous studies, it is suggested that Synechococcus SP. PCC 7002 is able to metabolize sulfide compounds. We took advantage of the results in our last experiment and adjusted the concentration of DCMU to an appropriate degree. Since sulfide would become the main reducing energy for photoassimilation under the effect of DCMU, we believe the more sulfide concentration in the wells, the better cell growth would be observed.



The effect of sodium sulfide on Synechococcus SP. PCC 7942 growth rate

After thoroughly examined the ability of sqr in Synechococcus SP. PCC 7002, we planned to conduct a series of similar experiments on Synechococcus SP. PCC 7942. Except for the cultivation medium, other growing conditions remained the same. Instinctively, the strain expressing sqr should grow better than the wile type strain. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper.


Sulfide concentration and the growth of sqr expressing strain Synechococcus SP. PCC 7942

It was expected that SQR expressing strain and wild type counterpart would have different growth rate under the presence of sulfide compounds. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper. As the result, we analyze H2S amount to detect whether sqr gene work or not. Therefore, we perform Chemical microvolume turbidimetry method to detect H2S concentration (see Sulfur Oxide Terminator part)


Sulfide oxidation in Escherichia coli expressing sulfide-quinone reductase gene

Repots have it that Escherichia coli can express functional sulfide-quinone reductase (SQR). Therefore, we slightly adjusted the previous experiment and applied to the SQR gene from Synechococcus SP. PCC 7002. With methylene blue method, we would test the efficiency of SQR sulfide oxidation. Since such method involved in measurement of optical density, it is more appropriate to perform such experiment on colorless bacteria instead of engineered cyanobacteria strain.