Difference between revisions of "Team:UPMC PARIS/Parts"

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<h1>Parts</h1>
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    <h2 class="centered">Part submission : BBa_K2504000</h2>
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<p>Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The <code>&lt;groupparts&gt;</code> tag (see below) will generate a table with all of the parts that your team adds to your team sandbox.</p>
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<p>Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without needing to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.</p>
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      <h4>Introduction</h4>
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      <p>
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        The 17 kilodalton protein called Skp is a molecular chaperone that resides in the periplasm of Escherichia coli. The SKP protein folding enables the expression of high-level recombinant proteins which contributes to the development of therapeutic molecules to treat misfolded proteins in some diseases. Drug development of such molecules can lead to production with bacteria of anti-cancer vaccines. Consequently, Skp has been shown to improve expression of various scFvs, in addition to larger Fab fragments and intact immunoglobulins (Raphael Levy and all,2011). This improvement of bacterial strain by genomic integration could brings a robust biological tool related to our project and an alternative in the use of antibiotics. Other genomic integrations will be realized in the continuity of our project to avoid maintaining plasmids through antibiotics, and to facilitate the use of our factory. In a second time it could be also relevant to permit production of others helper proteins like GroEL/ES.
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      <br><br>
  
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      <h4>Achievements</h4>
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      <p>
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        We submitted our biobrick sequence to the iGEM community, and we documented the experimental characterization of this part on the Main Page of that Part’s Registry entry.
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      <br><br>
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      <h4>Method</h4>
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      <br>
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        We started from the cytosolic form of Skp under the control of T7 promoter, ended by a T7 terminator and flanked by iGEM prefix and suffix by synthesized by IDT.
  
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        <br><br>We used the NotI site to insert the sequence into the iGEM plasmid pSB1C3 and we transformed competent E. coli DH5α.
  
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        <br><br>After transformation, we performed a plasmid DNA extraction. We selected recombinante clone by PCR using VF and VF2 primers. We collaborate with sequencing platform Eurofin in order to verify our DNA sequence by sequencing and dry the remaining DNA to send it to the iGEM foundation.
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      <h4>Results</h4>
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<h5>Note</h5>
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<p>Note that parts must be documented on the <a href="http://parts.igem.org/Main_Page"> Registry</a>. This page serves to <i>showcase</i> the parts you have made. Future teams and other users and are much more likely to find parts by looking in the Registry than by looking at your team wiki.</p>
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      Figure 6 Agarose gel electrophoresis (1.5%) after migration of PCR products for the verification of the insertion the biobrick BBa_K2504000
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      <br><br><br><br>
  
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      The presence of a band at 695pb suggest that the integration in the pSB1C3 worked as expected. We submit our biobrick with n=5 and sand a sample to sequencing to ensure the quality of our biobricks.
  
 +
      <br><br>Thus our Biobrick encodes for a cytoplasmic form of the chaperones Seventeen kilodalton protein (Skp). The Biobrick construction is composed of a T7 promoter, the coding DNA sequence of Skp protein truncated by its localisation signal sequence and a T7 terminator.
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<h5>Adding parts to the registry</h5>
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<p>You can add parts to the Registry at our <a href="http://parts.igem.org/Add_a_Part_to_the_Registry">Add a Part to the Registry</a> link.</p>
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<p>We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better you will remember all the details about your parts. Remember, you don't need to send us the DNA sample before you create an entry for a part on the Registry. (However, you <b>do</b> need to send us the DNA sample before the Jamboree. If you don't send us a DNA sample of a part, that part will not be eligible for awards and medal criteria.)</p>
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<h5>What information do I need to start putting my parts on the Registry?</h5>
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        <br>
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<p>The information needed to initially create a part on the Registry is:</p>
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        <h3>Sponsors</h3>
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<ul>
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<li>Part Name</li>
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<li>Part type</li>
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<li>Creator</li>
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<li>Sequence</li>
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<li>Short Description (60 characters on what the DNA does)</li>
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<li>Long Description (Longer description of what the DNA does)</li>
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            <img height="30%" width="30%" src="sponsors/UPMC_team_2017_sponsors_Ambassade_france.jpg" alt="">
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<li>Design considerations</li>
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We encourage you to put up <em>much more</em> information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page. </p>
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<h5>Inspiration</h5>
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<p>We have a created  a <a href="http://parts.igem.org/Well_Documented_Parts">collection of well documented parts</a> that can help you get started.</p>
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<p> You can also take a look at how other teams have documented their parts in their wiki:</p>
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<li><a href="https://2014.igem.org/Team:MIT/Parts"> 2014 MIT </a></li>
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<li><a href="https://2014.igem.org/Team:Heidelberg/Parts"> 2014 Heidelberg</a></li>
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            <img height="30%" width="30%" src="sponsors/UPMC_team_2017_sponsors_NEB.png" alt="">
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<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Parts">2014 Tokyo Tech</a></li>
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<h5>Part Table </h5>
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<p>Please include a table of all the parts your team has made during your project on this page. Remember part characterization and measurement data must go on your team part pages on the Registry. </p>
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            <img src="sponsors/UPMC_team_2017_sponsors_CFA_UPMC.png" alt="">
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<groupparts>iGEM17 UPMC_PARIS</groupparts>
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            <img height="100%" width="100%" src="sponsors/UPMC_team_2017_sponsors_MAster_BMC.jpg" alt="">
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            <img height="100%" width="100%" src="sponsors/UPMC_team_2017_sponsors_UFR_bio.jpg" alt="">
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Revision as of 18:28, 24 November 2017

Impact UPMC



Part submission : BBa_K2504000

Introduction

The 17 kilodalton protein called Skp is a molecular chaperone that resides in the periplasm of Escherichia coli. The SKP protein folding enables the expression of high-level recombinant proteins which contributes to the development of therapeutic molecules to treat misfolded proteins in some diseases. Drug development of such molecules can lead to production with bacteria of anti-cancer vaccines. Consequently, Skp has been shown to improve expression of various scFvs, in addition to larger Fab fragments and intact immunoglobulins (Raphael Levy and all,2011). This improvement of bacterial strain by genomic integration could brings a robust biological tool related to our project and an alternative in the use of antibiotics. Other genomic integrations will be realized in the continuity of our project to avoid maintaining plasmids through antibiotics, and to facilitate the use of our factory. In a second time it could be also relevant to permit production of others helper proteins like GroEL/ES.



Achievements

We submitted our biobrick sequence to the iGEM community, and we documented the experimental characterization of this part on the Main Page of that Part’s Registry entry.



Method


We started from the cytosolic form of Skp under the control of T7 promoter, ended by a T7 terminator and flanked by iGEM prefix and suffix by synthesized by IDT.

We used the NotI site to insert the sequence into the iGEM plasmid pSB1C3 and we transformed competent E. coli DH5α.

After transformation, we performed a plasmid DNA extraction. We selected recombinante clone by PCR using VF and VF2 primers. We collaborate with sequencing platform Eurofin in order to verify our DNA sequence by sequencing and dry the remaining DNA to send it to the iGEM foundation.


Results


Figure 6 Agarose gel electrophoresis (1.5%) after migration of PCR products for the verification of the insertion the biobrick BBa_K2504000



The presence of a band at 695pb suggest that the integration in the pSB1C3 worked as expected. We submit our biobrick with n=5 and sand a sample to sequencing to ensure the quality of our biobricks.

Thus our Biobrick encodes for a cytoplasmic form of the chaperones Seventeen kilodalton protein (Skp). The Biobrick construction is composed of a T7 promoter, the coding DNA sequence of Skp protein truncated by its localisation signal sequence and a T7 terminator.


Sponsors