Difference between revisions of "Team:Shanghai HS ID/Results"

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<h1>Results</h1>
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<p>You can describe the results of your project and your future plans here. </p>
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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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<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 <b>all measurement and characterization data must also be on the Part's Main Page on the <a href="http://parts.igem.org/Main_Page">Registry</a>.</b> Otherwise these data will not be in consideration for any medals or part awards! </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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<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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<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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<ul>
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<li><a href="https://2019.igem.org/Team:Newcastle/Results">2019 Newcastle</a></li>
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<li><a href="https://2019.igem.org/Team:Munich/Results">2019 Munich </a></li>
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<li><a href="https://2019.igem.org/Team:Tec-Chihuahua/Results">2019 Tec Chihuahua</a></li>
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<li><a href="https://2020.igem.org/Team:Aalto-Helsinki/Results">2020 Aalto Helsinki</a></li>
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<li><a href="https://2020.igem.org/Team:GreatBay_SCIE/Results">2020 GreatBay SCIE</a></li>
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<li><a href="https://2020.igem.org/Team:Queens_Canada/Results">2020 Queens Canada</a></li>
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        <div class="sub-title">Results</div>
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            1. DNA Profiles
 +
        </div>
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        <div class="img-wrap no-margin">
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            <img src="https://static.igem.org/mediawiki/2021/5/54/T--Shanghai_HS_ID--results01.jpg" alt="" />
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            <span>Figure 1. DNA profile of the plasmid PLCNICK</span>
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        </div>
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        <div class="img-wrap no-margin">
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            <img src="https://static.igem.org/mediawiki/2021/c/c7/T--Shanghai_HS_ID--results02.jpg" alt="" />
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            <span>Figure 2. DNA Profile of plasmid PTB165 which was electrotransformed into L. casei for transformation
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                efficiency test</span>
 +
        </div>
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        <div class="img-wrap no-margin">
 +
            <img src="https://static.igem.org/mediawiki/2021/0/0f/T--Shanghai_HS_ID--results03.jpg" alt="" />
 +
            <span> Figure 3. DNA Profile of LSEI-2094+ upstream and downstream homologous arms</span>
 +
        </div>
 +
        <div class="article-title">2. Electrophoresis result after PCR</div>
 +
        <div class="img-wrap no-margin">
 +
            <img src="https://static.igem.org/mediawiki/2021/a/a8/T--Shanghai_HS_ID--results04.jpg" alt="" />
 +
            <span>Figure 4. Electrophoretogram of pLCNICK PCR resiult</span>
 +
        </div>
 +
        <div class="web info">1-4 are pLCNICK after enzyme digestion, 5 is pLCNICK plasmid before enzyme digestion, 6~7
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            are upstream
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            homologous arms after PCR, 8~9 are downstream homologous arms after PCR.</div>
 +
        <div class="article-title">3. Verification result of colony PCR</div>
 +
        <div class="img-wrap no-margin">
 +
            <img src="https://static.igem.org/mediawiki/2021/2/29/T--Shanghai_HS_ID--results05.jpg" alt="" />
 +
            <span>Figure 5. PCR verification result</span>
 +
        </div>
 +
        <div>As showing above, there are bands (1~3, 6, 8~12) at 1000 bp around which are consistent with the DNA
 +
            profile of the downstream homologous arms. Therefore, it indicated that we have successfully transformed the
 +
            plasmid in E. coli.</div>
 +
        <div class="article-title">4. Transformation test result</div>
 +
        <div class="img-wrap no-margin">
 +
            <img src="https://static.igem.org/mediawiki/2021/b/b9/T--Shanghai_HS_ID--results06.jpg" alt="" />
 +
            <span>Graph 1. Comparison between the wild L. caseil and modified L. casei in transformation</span>
 +
        </div>
 +
        <div class="article-content">After we electrotransformed the plasmid pIB165 as the foreign DNA to test the
 +
            transformation efficiency of our modified L. casei, it took several days to culture and finally saw the
 +
            comparison results. As showing above, we can see that the modified L. casei (KO) has higher transformation
 +
            efficiency with remarkably more colonies than the wild (Wild).</div>
 +
        <div class="img-wrap no-margin">
 +
            <img src="https://static.igem.org/mediawiki/2021/d/d8/T--Shanghai_HS_ID--results07.jpg" alt="" />
 +
            <span>Figure 6. Histogram comparison between wild strain and KO strain</span>
 +
        </div>
  
 +
        <div class="article-content">
 +
            In addition, we measured OD<sub>600</sub> of these strain groups which were pre-spreaded plates with
 +
            different volumes of bacteria solutions so as to quantify the transformation results as showing above (Fig.
 +
            6).<br />
 +
            In conclusion, it indicates that our modified L. casei has much higher efficiency of the foreign plasmid
 +
            transformation than the wild and the modified L. casei has great potential to be used as the recombinant
 +
            carrier in various areas.
 +
        </div>
 +
        <div class="article-title">5. Future plan and our concern</div>
 +
        <div class="article-content">
 +
            As we have primarily tested the transformation effiency of our modified L. casei, we would think about to
 +
            further explore the expression performance of the foreign plasmid as well. Besides, we have built the
 +
            partnership with EV71 Terminator (2021 iGEM team Shanghai_Metropolis ) who we will work together to develop
 +
            a HFMD Oral Vaccines using L. casei as the live carrier. Therefore, in next step we will electrotransform
 +
            the plasmid constructed by EV71 Terminator into our modified L. casei and test the expression performance of
 +
            this recombinant L. casei.<br />
 +
            About our modified L. casei as well as its derived recombinant bacterium, we also take the safety and
 +
            stability into acount, it would be important for us to do more literature research and seek more
 +
            professional advice on future development of our product before we step into the real application.
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Revision as of 16:49, 14 October 2021

Shanghai_HS_ID

Results
1. DNA Profiles
Figure 1. DNA profile of the plasmid PLCNICK
Figure 2. DNA Profile of plasmid PTB165 which was electrotransformed into L. casei for transformation efficiency test
Figure 3. DNA Profile of LSEI-2094+ upstream and downstream homologous arms
2. Electrophoresis result after PCR
Figure 4. Electrophoretogram of pLCNICK PCR resiult
1-4 are pLCNICK after enzyme digestion, 5 is pLCNICK plasmid before enzyme digestion, 6~7 are upstream homologous arms after PCR, 8~9 are downstream homologous arms after PCR.
3. Verification result of colony PCR
Figure 5. PCR verification result
As showing above, there are bands (1~3, 6, 8~12) at 1000 bp around which are consistent with the DNA profile of the downstream homologous arms. Therefore, it indicated that we have successfully transformed the plasmid in E. coli.
4. Transformation test result
Graph 1. Comparison between the wild L. caseil and modified L. casei in transformation
After we electrotransformed the plasmid pIB165 as the foreign DNA to test the transformation efficiency of our modified L. casei, it took several days to culture and finally saw the comparison results. As showing above, we can see that the modified L. casei (KO) has higher transformation efficiency with remarkably more colonies than the wild (Wild).
Figure 6. Histogram comparison between wild strain and KO strain
In addition, we measured OD600 of these strain groups which were pre-spreaded plates with different volumes of bacteria solutions so as to quantify the transformation results as showing above (Fig. 6).
In conclusion, it indicates that our modified L. casei has much higher efficiency of the foreign plasmid transformation than the wild and the modified L. casei has great potential to be used as the recombinant carrier in various areas.
5. Future plan and our concern
As we have primarily tested the transformation effiency of our modified L. casei, we would think about to further explore the expression performance of the foreign plasmid as well. Besides, we have built the partnership with EV71 Terminator (2021 iGEM team Shanghai_Metropolis ) who we will work together to develop a HFMD Oral Vaccines using L. casei as the live carrier. Therefore, in next step we will electrotransform the plasmid constructed by EV71 Terminator into our modified L. casei and test the expression performance of this recombinant L. casei.
About our modified L. casei as well as its derived recombinant bacterium, we also take the safety and stability into acount, it would be important for us to do more literature research and seek more professional advice on future development of our product before we step into the real application.
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