Difference between revisions of "Team:CHINA CD UESTC/Results"
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<h3>Part 1. Laccase</h3> | <h3>Part 1. Laccase</h3> | ||
<p> | <p> | ||
− | After our experiments, we successfully constructed the vector piGEM-R-Lac which combined Laccase with RFP by fusion expression. We measured different OD <sub>600</sub> | + | After our experiments, we successfully constructed the vector piGEM-R-Lac which combined Laccase with RFP by fusion expression. We measured different OD<sub>600</sub> of bacterium at different times, and record the color. We also detected the activity of laccase. We found that the activity of Laccase has positive correlation with OD<sub>600</sub> and the color. We made sure that the activity of Laccase have the catalytic property by using ABTS method. So that we could put them on cathode of EBFC and make it work. |
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<div class="clear"></div> | <div class="clear"></div> | ||
− | <p> <strong>(1)SDS-PAGE</strong> | + | <p> |
+ | <strong>(1)SDS-PAGE</strong> | ||
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<div class="project_pic"> | <div class="project_pic"> | ||
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<img src="https://static.igem.org/mediawiki/2015/2/2f/CHINA_CD_UESTC_RESULT01.jpg" width="40%" style="margin-left:100px;"> | <img src="https://static.igem.org/mediawiki/2015/2/2f/CHINA_CD_UESTC_RESULT01.jpg" width="40%" style="margin-left:100px;"> | ||
<p id="pic_illustration"> | <p id="pic_illustration"> | ||
− | Lane1: Control. Lane2: Induced expression (BL21). | + | Lane1: Control. Lane2: Induced expression (BL21). <br>Induction conditions: 37centigrades,200rpm,induce 10 hours with IPTG which final concentration is 0.5mM. Using Ultrasonic Cell Disruptor to crush the bacterium in ice-bath. |
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</p> | </p> | ||
<div class="clear"></div> | <div class="clear"></div> | ||
− | <p> <strong>(2)Bacterium liquid</strong> | + | <p> |
+ | <strong>(2)Bacterium liquid</strong> | ||
</p> | </p> | ||
<p>concentration of bacterium detected at different time</p> | <p>concentration of bacterium detected at different time</p> | ||
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<img class="left_pic" src="https://static.igem.org/mediawiki/2015/6/69/CHINA_CD_UESTC_RESULT02.jpg" width="380px" height="270px"> | <img class="left_pic" src="https://static.igem.org/mediawiki/2015/6/69/CHINA_CD_UESTC_RESULT02.jpg" width="380px" height="270px"> | ||
<img class="right_pic" src="https://static.igem.org/mediawiki/2015/3/3d/CHINA_CD_UESTC_RESULT03.png" width="380px" height="270px"></div> | <img class="right_pic" src="https://static.igem.org/mediawiki/2015/3/3d/CHINA_CD_UESTC_RESULT03.png" width="380px" height="270px"></div> | ||
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<strong>(3)Supernatant after crush:</strong> | <strong>(3)Supernatant after crush:</strong> | ||
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Using Ultrasonic Cell Disruptor to crush the bacterium in ice-bath. Collect the Supernatant and detect the activity of Laccase by using ABTS. The conclusion is that the activity of Laccasse is increasing with time passing by. | Using Ultrasonic Cell Disruptor to crush the bacterium in ice-bath. Collect the Supernatant and detect the activity of Laccase by using ABTS. The conclusion is that the activity of Laccasse is increasing with time passing by. | ||
</p> | </p> | ||
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− | < | + | <img class="left_pic" src="https://static.igem.org/mediawiki/2015/8/84/CHINA_CD_UESTC_RESULT04.jpg" width="380px" height="270px"> |
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<h3>Part2. Vectors construction</h3> | <h3>Part2. Vectors construction</h3> | ||
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<p id="pic_title">This is part has relation with the formation of magnetosome</p> | <p id="pic_title">This is part has relation with the formation of magnetosome</p> | ||
<img src="https://static.igem.org/mediawiki/2015/c/c0/CHINA_CD_UESTC_RESULT10.png" width="60%"> | <img src="https://static.igem.org/mediawiki/2015/c/c0/CHINA_CD_UESTC_RESULT10.png" width="60%"> | ||
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<p id="pic_illustration"></p> | <p id="pic_illustration"></p> | ||
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<h3>Part 3. Vectors table</h3> | <h3>Part 3. Vectors table</h3> | ||
<p> | <p> | ||
− | In order to improve the catalytic efficiency of laccase, we decided to immobilize this enzyme. After looking over lots of methods to immobilize laccase. We finally decide to bind laccase to magnetosome. To bind laccase to magnetosome, we designed a series of experiments to let Escherichia coli to express the proteins which are related with the information of the magnetosome, and to get magnetosomes. Through three mouth, we succeed to construct fifteen recombinant plasmids. Details are as follows: | + | In order to improve the catalytic efficiency of laccase, we decided to immobilize this enzyme. After looking over lots of methods to immobilize laccase. We finally decide to bind laccase to magnetosome. To bind laccase to magnetosome, we designed a series of experiments to let Escherichia coli to express the proteins which are related with the information of the magnetosome, and to get magnetosomes. Through three mouth, we succeed to construct fifteen recombinant plasmids. Details are as follows: |
</p> | </p> | ||
− | <p> | + | <p> <strong>1.For expression</strong> |
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</p> | </p> | ||
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</tbody> | </tbody> | ||
</table> | </table> | ||
− | <p> | + | <p> <strong>2.For detection</strong> |
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</p> | </p> | ||
<table class="table1"> | <table class="table1"> | ||
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Revision as of 15:54, 13 September 2015
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RESULTS
We present details on the various methods such as vectors design, domain linker selection and choose of enzyme insertion site that used in the experiment on this page, if you are willing to check or follow our work, you can scan the methods here. Any questions or advice to us are acceptable at any time.