Difference between revisions of "Team:GZHS-United/Results"

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{{GZHS-United}}
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<html>
 
<html>
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<head>
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<title>result</title>
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<link href="https://2017.igem.org/Team:GZHS-United/CSS/bootstrapmin?action=raw&ctype=text/css" rel="stylesheet" type="text/css"/>
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<style>
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#GZHS_content img {
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<body>
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<div class="header-bottom">
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<nav class="cl-effect-5">
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<li role="presentation">
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Attributions">Attributions</a>
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<li role="presentation">
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Collaborations">Collaborations</a>
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<span data-hover="PROJECT">PROJECT</span>
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Description">Description</a>
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<li role="presentation">
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Design">Design</a>
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<li role="presentation">
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Experiments">Experiments</a>
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</ul>
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<li class="dropdown">
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<span data-hover="RESULTS">RESULTS</span>
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Results">Results</a>
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<a role="menuitem" tabindex="-1" href="https://2017.igem.org/Team:GZHS-United/Entrepreneurship">Entrepreneurship</a>
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</ul>
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</nav>
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</ul>
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</div>
  
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<div class="column full_size" >
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</div>
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</div>
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<h1>Results</h1>
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<p>Here you can describe the results of your project and your future plans. </p>
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<h5>What should this page contain?</h5>
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<ul>
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<li> Clearly and objectively describe the results of your work.</li>
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<li> Future plans for the project. </li>
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<li> Considerations for replicating the experiments. </li>
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</ul>
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<h5>You should also describe what your results mean: </h5>
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<ul>
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<li> Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for. </li>
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<li> Show data, but remember all measurement and characterization data must be on part pages in the Registry. </li>
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<li> Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project. </li>
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</ul>
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</div>
 
</div>
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<div class="content" id="GZHS_content">
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<div class="clear"></div>
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<div class="container">  
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<div class="row" style="padding:60px 0;">
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<div class="column half_size" >
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<h1>Result</h1>
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<img src="https://static.igem.org/mediawiki/2017/2/2d/T--GZHS-United--results_top.png" class="top_img">
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<p>The first step of our project is genome DNA extraction, lane 1 is the genome DNA of <em>bacillus thuringiensis .Israelensis</em>, lane 2 is the genome DNA of <em>Bacillus sphaericus</em>. According to the gel, the length of the two genomes are correct. </p>
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<h5> Project Achievements </h5>
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<div class="result_img">
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<img src="https://static.igem.org/mediawiki/2017/e/e5/T--GZHS-United--result1.png">
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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</div>
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<p>After  genome DNA extraction, we used them as template to amplify Cry4Ba and Mtx1 by PCR, and the results of the amplification were shown in figure 2. </p>
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<ul>
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<div class="result_img">
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<li>A list of linked bullet points of the successful results during your project</li>
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<img src="https://static.igem.org/mediawiki/2017/a/a7/T--GZHS-United--result2.png">
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
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</div>
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</ul>
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<p>After the target genes were amplified, we have constructed our vector by linking target gene into pEASY-BluntE1 through topo cloning, then transferred the expression vector into <em>E. coli</em> BL21 for protein expression and purification. Figure 3 is the expression vector of <em>Cry4Ba</em> and the outcome after purification. According to the gel, lane 1 is our negative control containing bacterial protein of <em>E. coli</em> BL21, lane 2 is our target protein before purification containing bacterial protein of <em>Cry4Ba E.coli</em> BL21, lane 3 is the protein <em>Cry4Ba</em> after purification. It can be seen from the graph that we have successfully obtained the target protein with high purity.</p>
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<div class="row">
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<div class="col-sm-6 col-md-6 col-lg-6 result_img_left">
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<img src="https://static.igem.org/mediawiki/2017/1/12/T--GZHS-United--result3.png">
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</div>
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<div class="col-sm-6 col-md-6 col-lg-6 result_img_right">
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<img src="https://static.igem.org/mediawiki/2017/7/72/T--GZHS-United--result4.png">
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</div>
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</div>
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<p>To produce target protein in a large scale, we tried different induction time and IPTG concentration to find the optimum condition for <em>Cry4Ba</em> expression. It can be seen from figure 4 and 5 that the expressing quantity increased as the increasing of induction time, but the protein expression level has no correlation with IPTG concentration. Therefore, concerning the parameters we mentioned above, we choose to incubate for 6 hours with 0.25mM IPTG as our experimental procedure. </p>
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<div class="result_img">
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<img src="https://static.igem.org/mediawiki/2017/7/79/T--GZHS-United--result5.png">
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</div>
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</div>
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</div>
 
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</body>
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<div id="to_top">
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<div class="column half_size" >
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<img src="https://static.igem.org/mediawiki/2017/9/97/T--GZHS-United--toTop_buttom.png" onClick="toTop()"/>
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<h5>Inspiration</h5>
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<p>See how other teams presented their results.</p>
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<ul>
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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</ul>
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<div id="next_page">
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<a href="https://2017.igem.org/Team:GZHS-United/Model"><img src="https://static.igem.org/mediawiki/2017/b/bb/T--GZHS-United--right_buttom.png"/></a>
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Latest revision as of 17:41, 28 October 2017

result

Result

The first step of our project is genome DNA extraction, lane 1 is the genome DNA of bacillus thuringiensis .Israelensis, lane 2 is the genome DNA of Bacillus sphaericus. According to the gel, the length of the two genomes are correct.

After genome DNA extraction, we used them as template to amplify Cry4Ba and Mtx1 by PCR, and the results of the amplification were shown in figure 2.

After the target genes were amplified, we have constructed our vector by linking target gene into pEASY-BluntE1 through topo cloning, then transferred the expression vector into E. coli BL21 for protein expression and purification. Figure 3 is the expression vector of Cry4Ba and the outcome after purification. According to the gel, lane 1 is our negative control containing bacterial protein of E. coli BL21, lane 2 is our target protein before purification containing bacterial protein of Cry4Ba E.coli BL21, lane 3 is the protein Cry4Ba after purification. It can be seen from the graph that we have successfully obtained the target protein with high purity.

To produce target protein in a large scale, we tried different induction time and IPTG concentration to find the optimum condition for Cry4Ba expression. It can be seen from figure 4 and 5 that the expressing quantity increased as the increasing of induction time, but the protein expression level has no correlation with IPTG concentration. Therefore, concerning the parameters we mentioned above, we choose to incubate for 6 hours with 0.25mM IPTG as our experimental procedure.