Team:OUC-China/Results

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  <meta name="description" content="">
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<title>Results</title>
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<title>Overview</title>
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      <p>OUC-China IGEM 2013</p>
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                <a class="dropdown-toggle" id="drop4" role="button" data-toggle="dropdown" href="#"><font size="4.5px">Project</font><b class="caret"></b>
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<li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Overview">Overview</a>
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      <ul class="nav navbar-nav pull-right">
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                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Achievement">Achievement & judge criteria</a>
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      <li><a href="https://2013.igem.org/Team:OUC-China">Home</a></li>
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                  <a tabindex="-1" href="#">Intercellular compartment</a>
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                      <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Review">Review</a></li>
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          <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Overview">Overview</a></li>
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                      <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Design">Design</a></li>
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                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Achievement">Achievement & judge criteria</a></li>
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  <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Microfluidics">Microfluidics</a></li>
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                    <li class="dropdown-submenu">
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  <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Results">Results</a></li>
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                      <a tabindex="-1" href="#">Intracellular compartment</a>
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                        <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Review">Review</a></li>
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  </li>
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                        <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Design">Design</a></li>
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              <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Microfluidics">Microfluidics</a></li>
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              <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Results">Results</a></li>
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                   <a tabindex="-1" href="#">RNA guardian</a>
                   <a tabindex="-1" href="#">RNA guardian</a>
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                       <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Instruction">Instruction</a></li>
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                       <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Introduction">Introduction</a></li>
                       <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/RNA guardian/Design">Design</a></li>
                       <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/RNA guardian/Design">Design</a></li>
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  <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/RNA guardian/Results">Result</a></li>
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            <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/RNA guardian/Results">Result</a></li>
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  <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Reference">Reference</a></li>
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            <li><a tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Reference">Reference</a></li>
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          <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Part description">Part description</a></li>
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          <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Future work">Future work</a></li>
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  <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Part description">Part description</a></li>
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  <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Future work">Future work</a></li>
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                 </ul>
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                <a class="dropdown-toggle" id="drop5" role="button" data-toggle="dropdown" href="#"><font size="4.5px">Modeling</font><b class="caret"></b></a>
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                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Magnetic Analysis">Magnetic Analysis</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Magnetic Analysis">Magnetic Analysis</a></li>
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                  <li role="presentation" class="divider"></li>
 
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Model in RNA guardian">Model in RNA guardian</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Model in RNA guardian">Model in RNA guardian</a></li>
                 </ul>
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            <li><a  href="https://2013.igem.org/Team:OUC-China/Human Practice">Human Practice</a></li>
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                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Members">Members</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Members">Members</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Instructors">Instructors</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Instructors">Instructors</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Advisor">Advisor</a></li>
                   <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Advisor">Advisor</a></li>
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  <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Lab">Lab</a></li>
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          <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Lab">Lab</a></li>
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  <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Acknowledgement">Acknowledgement</a></li>
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          <li role="presentation"><a role="menuitem" tabindex="-1" href="https://2013.igem.org/Team:OUC-China/Acknowledgement">Acknowledgement</a></li>
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      <li class="dropdown"><a  href="https://2013.igem.org/Team:OUC-China/Safety">Safety</a></li>
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           <p><img src="https://static.igem.org/mediawiki/2013/8/8b/Ouc-result1.jpg" width="400"/><br />Figure. 1 The laser confocal microscopy result of JM109 with no plasmid. There is no GFP gathering in view.<br /><img src="https://static.igem.org/mediawiki/2013/2/26/Ouc-result5.jpg" width="400"/><br />Figure. 2 The laser confocal microscopy results of JM109 with MamC::GFP expression plasmid(the left) and the JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster(the right).<br /><br />As we see there are obvious GFP signals generally equally distributed in JM109 with MamC::GFP expression plasmid, only a few bacteria contain the GFP gatherings at both ends of cells. And there no GFP signals in middle of the cell. There are GFP gatherings at both ends of cells. Compared with the JM109 with MamC::GFP expression plasmid, the figure(on the right) of JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster shows that there are clear MamC::GFP fluorescence gatherings at both ends and in middle area of the cell, which is different from the results of the other two samples. Additionally, we observed that the shape of the strain with both plasmids has changed. It has become much longer than normal <i>E.coli</i>, as the figure you can see(on the right). And the length of strain with both plasmids is almost three times longer than normal <i>E.coli.</i><br /> <br />We cannot be completely certain that the protein is actually attached to the membrane as we assumed, because the mechanism of the membrane formation is still not studied thoroughly. We cannot do TEM for the engineered bacteria at the moment due to forces beyond our control. You are welcome to follow us in our future work!
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      <div class="span9"><p style="font-weight:normal;"><font size="2px">Now we come to the most exciting part, our result. In this part we will show you our intracellular compartment in E.coli with laser confocal microscopy.<br />We detected three samples in total: The JM109 strain, the JM109 strain with MamC::GFP expression plasmid, and the JM109 strain with double MamC::GFP expression plasmid and our artificial gene cluster plasmid.<br /><br />Firstly, the result of JM109. Just as we see, there is only some noise and no obvious GFP in view.<br /><br /><img src="https://static.igem.org/mediawiki/2013/8/8b/Ouc-result1.jpg" height="500" width="600" /><br />Figure. 1 <br />The laser confocal microscopy result of JM109 with no plasmid. There is no GFP gathering in view.</font></p>
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  <div class="span9"><p style="font-weight:normal;"><font size="2px">Thirdly, the result of the JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster. The figure shows that there are clearly MamC::GFP fluorescence gatherings at both ends and in middle area of cell, which is different from the result of two samples above. <br /><br /><img src="https://static.igem.org/mediawiki/2013/c/cb/Ouc-result3.jpg" height="500" width="600"  /><br />Figure. 3<br />Additionally, we observed that the form of the strain with both plasmids has changed. It has become much longer than normal E.coli, like the figure below.<br /><br /><img src="https://static.igem.org/mediawiki/2013/6/65/Ouc-result4.jpg" height="500" width="600"  /><br />Figure. 4<br />The form of JM109 bacteria strain with both MamC::GFP expression plasmid under the 400X light microscope multiplication. The length of bacteria cell has become almost three times longer than normal <i>E.coli.</i></font></p></div>
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  <div class="span9"><p style="font-weight:normal;"><font size="2px"><b>About Dectection:</b><br />Our samples were all cultured in tubes with 20 mL LB medium in 37℃ in 170rpm incubator for 10h. Then we concentrated 1ml samples into 10μL, smeared it onto slides, sealed the slides, and observed slides using laser confocal microscopy (arouse wavelength is 488nm).</font></p></div>
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Latest revision as of 04:04, 28 September 2013

Overview


Figure. 1 The laser confocal microscopy result of JM109 with no plasmid. There is no GFP gathering in view.

Figure. 2 The laser confocal microscopy results of JM109 with MamC::GFP expression plasmid(the left) and the JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster(the right).

As we see there are obvious GFP signals generally equally distributed in JM109 with MamC::GFP expression plasmid, only a few bacteria contain the GFP gatherings at both ends of cells. And there no GFP signals in middle of the cell. There are GFP gatherings at both ends of cells. Compared with the JM109 with MamC::GFP expression plasmid, the figure(on the right) of JM109 strain with both MamC::GFP expression plasmid and artificial gene cluster shows that there are clear MamC::GFP fluorescence gatherings at both ends and in middle area of the cell, which is different from the results of the other two samples. Additionally, we observed that the shape of the strain with both plasmids has changed. It has become much longer than normal E.coli, as the figure you can see(on the right). And the length of strain with both plasmids is almost three times longer than normal E.coli.

We cannot be completely certain that the protein is actually attached to the membrane as we assumed, because the mechanism of the membrane formation is still not studied thoroughly. We cannot do TEM for the engineered bacteria at the moment due to forces beyond our control. You are welcome to follow us in our future work!

Retrieved from "http://2013.igem.org/Team:OUC-China/Results"