Difference between revisions of "Team:SDU-CHINA/Improve"

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{{SDU-CHINA}}
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<div class="column full_size judges-will-not-evaluate">
 
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<h3>★  ALERT! </h3>
 
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
 
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
 
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<h1>Improve</h1>
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<p>For teams seeking to improve upon a previous part or project, you should document all of your work on this page. Please remember to include all part measurement and characterization data on the part page on the Registry. Please include a link to your improved part on this page.</p>
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<h3>Gold Medal Criterion #2</h3>
 
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<p><b>Standard Tracks:</b> Create a new part that has a functional improvement upon an existing BioBrick part. The sequences of the new and existing parts must be different. You must perform experiments with both parts to demonstrate this improvement.  Document the experimental characterization on the Part's Main Page on the Registry for both the existing and new parts. Both the new and existing Main Page of each Part’s Registry entry must reference each other. Submit a sample of the new part to the Registry.
 
  
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The existing part must NOT be from your 2018 part number range and must be different from the part documented in bronze #4.
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<b>Special Tracks:</b> Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.</p>
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<body>
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<div class="body">
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    <div id="header">
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    <!--LOGO LINK--->
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    <div id="logo"><a href="https://2018.igem.org/Team:SDU-CHINA"><img src="https://static.igem.org/mediawiki/2018/7/72/T--SDU-China--duihui.jpg" alt="LOGO" /></a></div>
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                <a href="#">PROJECT &#9662;</a>
 +
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 +
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Description">Description</a></li>
 +
                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Design">Design</a></li>
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Results">Results</a></li>
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Demonstrate">Demonstrate</a></li>
 +
                </ul>
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                <a href="#">WETLAB &#9662;</a>
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 +
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Experiments">Protocols</a></li>
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Attributions">Attributoins</a></li>
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Collaborations">Collaborations</a></li>
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            <li><a href="https://2018.igem.org/Team:SDU-CHINA/InterLab">INTERLAB</a></li>
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            <li><a href="#">PARTS &#9662;</a>
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Parts">Parts</a></li>
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 +
                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Improve">Improve</a></li>
 +
                   
 +
                </ul>
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            <li><a href="https://2018.igem.org/Team:SDU-CHINA/Human_Practices">HUMAN PRACTICES &#9662;</a>
 +
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                    <!--links-->
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                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Human_Practices#Silver">Silver</a></li>
 +
                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Human_Practices#integrated">Gold & Integrated</a></li>
 +
                    <li><a href="https://2018.igem.org/Team:SDU-CHINA/Public_Engagement">Engagement</a></li>
 +
                </ul>
 +
                </li>
 +
        </ul>
 +
    </div>
 
</div>
 
</div>
  
 +
<div id="section">
 +
    <div class="conteudo">
 +
      <br> <h1 id="main-title">Improve</h1> <br>
 +
<div class="paragraphs">
 +
  <div class="bba"  style="text-align: center;"><h4><b><a href="http://parts.igem.org/Part:BBa_K2627000">BBa_K2627000</a></b></h4></div>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;This year we made an improvement on part <a href="http://parts.igem.org/Part:BBa_K592001">BBa_K592001</a> , which codes for green/red light sensing protein CcaS. CcaS functions in CcaS-CcaR two-component system responding to the existence of green or red light. CcaS consists of the following domains and regions: a N-terminal transmembrane helix, a sensor domain consisting of a cyanobacteriochrome-type cyclic guanosine monophosphate phosphodiesterase/adenylyl cyclase/formate hydrogen lyase transcriptional activator (GAF) domain, a linker region, two period/aryl hydrocarbon receptor nuclear translocator/single-minded (PAS) domains of unknown function, a second linker region, and a C-terminal HK domain (Fig. 1) <a href="#r1"><sup>[1]</sup></a>. Previously, the functions of some of the domains were studied, for example, a shape change can be observed in the GFA domain after CcaS was exposed to light, which could thus induce an autophosphorylation of the HK domain, then the phosphate group would be transferred to the regulator protein CcaR<a href="#r2"><sup>[2]</sup></a>. However, the function of PSA domains is remained unknown.
 +
<br></p>
 +
<img src="https://static.igem.org/mediawiki/2018/8/85/T--SDU-China--improve.png" alt=""> <br>
 +
<p class="reference"><b>Figure 1.</b> A brief illustration of the structure of CcaS. TF domain is located in the N-terminal while HK domain shows where the C-terminal is.</p>
 +
  <br>
 +
<p>&nbsp;&nbsp;&nbsp;&nbsp;We were inspired by Professor Koji’s work[1] in which he removed the two PSA domains to see what would happen. In his work, he created 11 variants of CcaS by deleting different numbers of base pairs, and these variants were characterized through measuring fluorescence/OD. We repeated his work and acquired this new part. This improved part is #4 in the variants collection. We found that this new part has a totally opposite response to green and red light compared with the original CcaS. As we can see from the figure below, #4 has a high fluorescence/Abs600 under red light but has a low one under green light.</p>
 +
<img src="https://static.igem.org/mediawiki/2018/2/26/T--SDU-China--improve2.png" alt=""> <br>
 +
<p class="reference"><b>Figure 2. </b>Characterization of part BBa_K2627000 and BBa_K592001. Part BBa_K2627000 shows a different effect after induced by green light and red light. Then fluorescence intensity of CcaS#4 was measured at 9h.</p>
 +
<br><br>
 +
<h4>References</h4>
 +
<p class="reference">
 +
[1].    Nakajima, M., et al., Construction of a Miniaturized Chromatic Acclimation Sensor from Cyanobacteria with Reversed Response to a Light Signal. Scientific Reports, 2016. 6(1).
 +
<br>
 +
[2].  Hirose, Y., Shimada, T., Narikawa, R., Katayama, M. & Ikeuchi M. Cyanobacteriochrome CcaS is the green light receptor that
 +
induces the expression of phycobilisome linker protein. Proc. Natl. Acad. Sci. USA 105, 9528–9533 (2008).
 +
</p>
 +
<!--<img src="https://static.igem.org/mediawiki/2018/e/e2/T--SDU-China--chara.png" alt="">-->
 +
</div>
 +
 +
        </div>
 +
    </div>
 +
 +
<div id="footer">
 +
    <div id="sponsors">
 +
        <img src="https://static.igem.org/mediawiki/2018/a/ab/T--SDU-CHINA--Sponsors0.svg" alt="SPONSORS" usemap="#Map-Sponsors" />
 +
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 +
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 +
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    <div id="contact">
 +
        <div><a href="mailto:sdubioigem@163.com">mailto:sdubioigem@163.com</a></div>
 +
        <div><a href="https://www.instagram.com/sdubiochina_2018/">www.instagram.com/sdubiochina_2018/</a></div>
 +
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Latest revision as of 03:58, 18 October 2018


Improve


BBa_K2627000

    This year we made an improvement on part BBa_K592001 , which codes for green/red light sensing protein CcaS. CcaS functions in CcaS-CcaR two-component system responding to the existence of green or red light. CcaS consists of the following domains and regions: a N-terminal transmembrane helix, a sensor domain consisting of a cyanobacteriochrome-type cyclic guanosine monophosphate phosphodiesterase/adenylyl cyclase/formate hydrogen lyase transcriptional activator (GAF) domain, a linker region, two period/aryl hydrocarbon receptor nuclear translocator/single-minded (PAS) domains of unknown function, a second linker region, and a C-terminal HK domain (Fig. 1) [1]. Previously, the functions of some of the domains were studied, for example, a shape change can be observed in the GFA domain after CcaS was exposed to light, which could thus induce an autophosphorylation of the HK domain, then the phosphate group would be transferred to the regulator protein CcaR[2]. However, the function of PSA domains is remained unknown.


Figure 1. A brief illustration of the structure of CcaS. TF domain is located in the N-terminal while HK domain shows where the C-terminal is.


    We were inspired by Professor Koji’s work[1] in which he removed the two PSA domains to see what would happen. In his work, he created 11 variants of CcaS by deleting different numbers of base pairs, and these variants were characterized through measuring fluorescence/OD. We repeated his work and acquired this new part. This improved part is #4 in the variants collection. We found that this new part has a totally opposite response to green and red light compared with the original CcaS. As we can see from the figure below, #4 has a high fluorescence/Abs600 under red light but has a low one under green light.


Figure 2. Characterization of part BBa_K2627000 and BBa_K592001. Part BBa_K2627000 shows a different effect after induced by green light and red light. Then fluorescence intensity of CcaS#4 was measured at 9h.



References

[1]. Nakajima, M., et al., Construction of a Miniaturized Chromatic Acclimation Sensor from Cyanobacteria with Reversed Response to a Light Signal. Scientific Reports, 2016. 6(1).
[2]. Hirose, Y., Shimada, T., Narikawa, R., Katayama, M. & Ikeuchi M. Cyanobacteriochrome CcaS is the green light receptor that induces the expression of phycobilisome linker protein. Proc. Natl. Acad. Sci. USA 105, 9528–9533 (2008).