Difference between revisions of "Team:KUAS Korea/Results"

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<table id="intro-title">
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    <tbody>
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        <td class="align-middle text-center section-header"><h3>Results</h3></td>
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<div class="column full_size">
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<br><br>
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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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        <div class="container">
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                <h4><strong>1. Beta-glucosidase expression</strong></h4>
  
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              <div style="text-align:justify">
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                <ol><br>Cooperator expresses beta-glucosidase with constitutive promoter to degrade cellobiose around it. We had the cell stock of cooperator with pCEL vector containing beta-glucosidase and transformed it into E.coli BW25113. The medium plate contains cellobiose instead of glucose to check whether E.coli that we transformed expresses beta-glucosidase on its surface. The streaking side below shows that cooperator grows on cellobiose containing M9 minimal medium.
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<img src="https://static.igem.org/mediawiki/2018/0/0c/T--KUAS_Korea--BETAGlucosidase_expression.png" height="300px" style="float: center;"hspace="" vspace="10">
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<br>[Fig 1.] Cultured cooperator on M9 minimal medium with 0.48% cellobiose
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<h3>What should this page contain?</h3>
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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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                <h4><strong>2. GFP expression</strong></h4>
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<div style="text-align:justify">
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                <ol>Cheater expresses GFP when it obtains glucose. We had the cell stock of cheater with the vector encoding GFP as a reporter gene and transformed it firstly into E.coli DH5α and then to E.coli BW25113.
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<img src="https://static.igem.org/mediawiki/2018/4/48/T--KUAS_Korea--GFP_PART1.png" height="300px" style="float: center;" hspace="5" vspace="100"
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<br>[Fig 2.] Cultured cheater on M9 minimal medium with 0.48% glucose
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<img src="https://static.igem.org/mediawiki/2018/1/1b/T--KUAS_Korea--GFP_PART2.png" height="300px" style="float: center;" hspace="5" vspace="100">
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<br>[Fig 2-1.] Cultured cheater under fluorescence microscopy showing the GFP activity
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                <h4><strong>3. Co-culture</strong></h4>
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<h3>Describe what your results mean </h3>
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                <ol>As the cheater and cooperator reached OD600 1.8 in M9 minimal medium with glucose concentration 0.48%, we began co-culture in M9 minimal medium with cellobiose concentration 0.48%. Batch co-culture was conducted in 5ml of medium in ratio of cheater and cooperator 1:1, 1:2, 1:4, 2:1, 4:1. 1 was counted as 1ul and each medium was incubated for 1, 3 and 4 days. After each day, we streaked 100ul of the incubated medium on LB agar plate with ampicillin. However, expected bacteria did not grow at all after overnight incubation.
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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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<ol>We predicted possible causes of unsuccessful co-culture as follows; <ul>
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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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<li>Too little amount of bacteria inoculated in the medium</li>
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</ul>
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<li>Low survival rate of pre-cultured bacteria since the optimal OD600 was 1.0 and ours was 1.8 which is thought to be after stationary phase.</li>  
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<li>Too short period of incubation</li><br>
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We plan to conduct the experiment once again with newly grown bacteria. 
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<h3> Project Achievements </h3>
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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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<ul>
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<li>A list of linked bullet points of the successful results during your project</li>
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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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<h3>Inspiration</h3>
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<p>See how other teams presented their results.</p>
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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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Latest revision as of 00:52, 18 October 2018

Results


1. Beta-glucosidase expression


    Cooperator expresses beta-glucosidase with constitutive promoter to degrade cellobiose around it. We had the cell stock of cooperator with pCEL vector containing beta-glucosidase and transformed it into E.coli BW25113. The medium plate contains cellobiose instead of glucose to check whether E.coli that we transformed expresses beta-glucosidase on its surface. The streaking side below shows that cooperator grows on cellobiose containing M9 minimal medium.


[Fig 1.] Cultured cooperator on M9 minimal medium with 0.48% cellobiose



2. GFP expression

    Cheater expresses GFP when it obtains glucose. We had the cell stock of cheater with the vector encoding GFP as a reporter gene and transformed it firstly into E.coli DH5α and then to E.coli BW25113.


[Fig 2.] Cultured cheater on M9 minimal medium with 0.48% glucose



[Fig 2-1.] Cultured cheater under fluorescence microscopy showing the GFP activity




3. Co-culture

    As the cheater and cooperator reached OD600 1.8 in M9 minimal medium with glucose concentration 0.48%, we began co-culture in M9 minimal medium with cellobiose concentration 0.48%. Batch co-culture was conducted in 5ml of medium in ratio of cheater and cooperator 1:1, 1:2, 1:4, 2:1, 4:1. 1 was counted as 1ul and each medium was incubated for 1, 3 and 4 days. After each day, we streaked 100ul of the incubated medium on LB agar plate with ampicillin. However, expected bacteria did not grow at all after overnight incubation.

    We predicted possible causes of unsuccessful co-culture as follows;
    • Too little amount of bacteria inoculated in the medium
    • Low survival rate of pre-cultured bacteria since the optimal OD600 was 1.0 and ours was 1.8 which is thought to be after stationary phase.
    • Too short period of incubation

      • We plan to conduct the experiment once again with newly grown bacteria.





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About Us

Student Leader: Taehoon Lee
Wet Lab Leader: Bosun Ryu
Dry Lab Leader: Uijin Han
Human Practices Leader: Eunmin Kim

Contact Us

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Tel: 02)1234-5678
Email: kuaskorea@gmail.com

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