Difference between revisions of "Team:TCFSH Taiwan/Composite Part"

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{{TCFSH_Taiwan}}
 
{{TCFSH_Taiwan}}
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{{TCFSH_Taiwan/Navigation}}
 
 
<html>
 
<html>
 
<head>
 
<head>
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     line-height:1;
 
     line-height:1;
 
}
 
}
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h1{
 
−
background-image:url('https://static.igem.org/mediawiki/2017/a/a9/Tcfshtaiwan_background.jpeg');
 
−
}
 
−
 
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.bigphoto{
 
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      margin: auto;
 
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/*大圖*/
 
/*大圖*/
 
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@media screen and (min-width: 768px) {
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.line .red {
 
.line .red {
 
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   height:47%; /*line寬度*/
 
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.line .blue {
 
.line .blue {
 
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   height: 20%;  /*調line長度*/
 
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     width:60vw;
 
     height:auto;
 
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.title{
 
.title{
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.picture{
 
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           <div class="node">
 
           <div class="node">
−
           <div class="topic"><p class="text_color">Basic Parts</p></div>
+
           <div class="topic"><p class="text_color">Composite Parts</p></div>
 +
            <div class="active-circle">
 +
              <div class="splash"></div>
 +
              <div class="cover">   
 +
              </div>
 +
            </div>
 +
            <div class="inactive-circle">
 +
              <div class="small-cover">
 +
              </div>
 +
            </div>
 +
          </div>
 +
 
 +
          <div class="node">
 +
          <div class="topic"><p class="text_color">PANTIDE without Lectin</p></div>
 +
            <div class="active-circle">
 +
              <div class="splash"></div>
 +
              <div class="cover">   
 +
              </div>
 +
            </div>
 +
            <div class="inactive-circle">
 +
              <div class="small-cover">
 +
              </div>
 +
            </div>
 +
          </div>
 +
 
 +
         
 +
          <div class="node">
 +
          <div class="topic"><p class="text_color">PANTIDE with Lectin</p></div>
 
             <div class="active-circle">
 
             <div class="active-circle">
 
               <div class="splash"></div>
 
               <div class="splash"></div>
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           </div>
 
           </div>
  
−
    </nav>
 
  
−
<h1>
+
          <div class="node">
 +
          <div class="topic"><p class="text_color">PANTIDE with GS Linker</p></div>
 +
            <div class="active-circle">
 +
              <div class="splash"></div>
 +
              <div class="cover">   
 +
              </div>
 +
            </div>
 +
            <div class="inactive-circle">
 +
              <div class="small-cover">
 +
              </div>
 +
            </div>
 +
          </div>
 +
         
 +
<div class="node">
 +
          <div class="topic"><p class="text_color">PANTIDE with GS Linker</p></div>
 +
            <div class="active-circle">
 +
              <div class="splash"></div>
 +
              <div class="cover">   
 +
              </div>
 +
            </div>
 +
            <div class="inactive-circle">
 +
              <div class="small-cover">
 +
              </div>
 +
            </div>
 +
          </div>
 +
 
 +
<div class="node">
 +
          <div class="topic"><p class="text_color">PANTIDE with GS Linker</p></div>
 +
            <div class="active-circle">
 +
              <div class="splash"></div>
 +
              <div class="cover">   
 +
              </div>
 +
            </div>
 +
            <div class="inactive-circle">
 +
              <div class="small-cover">
 +
              </div>
 +
            </div>
 +
          </div>
 +
 
 +
 +
    </nav>
  
 
<!--圖片-->
 
<!--圖片-->
 
   <div class="img-container">
 
   <div class="img-container">
−
         <img src="https://static.igem.org/mediawiki/2017/a/a9/Basicparts.jpeg" class="main-img" width="100%">
+
         <img src="https://static.igem.org/mediawiki/2016/9/98/NCTU_COMPOSITE_BIG_PICTURE.png
−
</div>
+
" class="main-img" width="100%">
−
 
+
  </div>
−
<!------------------wiki content--------------------->
+
  <div>
 +
    <img src='https://static.igem.org/mediawiki/2016/f/f8/Web.png' class="web img-position" height="200px" id="web">
 +
  </div>
 +
  <div>
 +
    <img src="https://static.igem.org/mediawiki/2016/6/65/Right_%281%29.png" class="right img-position" id="right" height="600px">
 +
  </div>
  
  
 +
<!--------------------wiki content----------------------->
 
<section class="content_container">
 
<section class="content_container">
 
<div>
 
<div>
 
<p class="title">Composite Parts</p>
 
<p class="title">Composite Parts</p>
−
<p class="content">We provide few basic parts which are used in our Detecoli, including<a href="http://parts.igem.org/Part:BBa_K2383001" class="biobrick">BBa_K2383001</a>and<a href="http://parts.igem.org/Part:BBa_K2383010" class="biobrick">BBa_K2383010</a>.</p>
+
<p class="content">We have known the function of the three toxic peptides of PANTIDE, Omega-hexatoxin-Hv1a, µ-segestritoxin-Sf1a, and Orally Active Insecticidal Peptide(OAIP) respectively and they are easy to express with the help of the bacteria <i>E. coli</i> BL21 Rosetta Gami strain. So, we then start to create BioBricks of these three toxins which we have obtained the nucleotide sequences. We use a strong T7 promoter that is suitable with the strain of <i>E. coli</i> we chose and also powerful at expressing proteins with multiple disulfide bonds, and we add His-tag for protein purification.</p>
  
 +
<p class="title">PANTIDE without Lectin</p>
 +
<p class="content"> PANTIDE can be a single toxic peptide of the three toxins we chose, Hv1a, Sf1a, and OAIP respectively, which derive from the natural predators spiders. </p>
  
−
<!-------cjBlue-LVA------>
+
<!----------Hv1a-------->
−
<a href="http://parts.igem.org/Part:BBa_K2383001" class="biobrick"><p class="content-1">BBa_K2383001</p></a>
+
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974011" class="biobrick"><p class="content-1">BBa_K1974011</p></a>
  
−
<img src="https://static.igem.org/mediawiki/2017/5/5e/Cjblue%2Blva.jpeg" class="bigphoto" width=70%;>
+
<div>
 +
      <img src="https://static.igem.org/mediawiki/2016/f/f7/NCTU_FORMOSA_H1.png" class="picture">
 +
</div>
  
−
<p class="content">This chromoprotein from the ''Cnidopus japonicus'' sea anemone, cjBlue, naturally exhibits dark green color when expressed. Compared to some other chromoproteins, such as amilCP (<partinfo>BBa_K592009</partinfo>), amilGFP (<partinfo>BBa_K592010</partinfo>), spisPink (<partinfo>BBa_K1033932</partinfo>), asPink (<partinfo>BBa_K1033933</partinfo>) and aeBlue (<partinfo>BBa_K864401</partinfo>), the color development is slower. The color is readily observed in both LB or on agar plates after 24-48 hours of incubation. The sequence is codon optimized for expression in ''E coli''. We add a LVA tag after it to accelerate its degradation rate.</p>
+
<p class="content">
 +
T7 promoter+RBS+Hv1a+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for Hv1a linked with His-tag.
 +
</p>
  
 +
<!---------Sf1a----------->
 +
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974012" class="biobrick"><p class="content-1">BBa_K1974012</p></a>
  
−
<!--------LacI----------->
 
−
<a href="http://parts.igem.org/Part:BBa_K2383010" class="biobrick"><p class="content-1">BBa_K2383010</p></a>
 
−
<p class="content">LacI without LVA tag</p>
 
 
<div>
 
<div>
 +
      <img src="https://static.igem.org/mediawiki/2016/a/ad/NCTU_FORMOSA_S1.png" class="picture">
 +
</div>
 +
 +
<p class="content">
 +
T7 promoter+RBS+Sf1a+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for Sf1a linked with His-tag.
 +
</p>
 +
 +
<!-------OAIP------------->
 +
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974013" class="biobrick"><p class="content-1">BBa_K1974013</p></a>
 +
 
<div>
 
<div>
−
       <img src="https://static.igem.org/mediawiki/2017/0/0d/LACInew.jpeg" class="bigphoto" width=75%;>
+
       <img src="https://static.igem.org/mediawiki/2016/2/21/NCTU_FORMOSA_O.png" class="picture">
 
</div>
 
</div>
 +
 +
<p class="content">
 +
T7 promoter+RBS+OAIP+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for OAIP linked with His-tag.
 +
</p>
 +
 +
 +
 +
<!---------------lectin---------------->
 +
 +
 +
 +
<p class="title">PANTIDE with Lectin</p>
 +
<p class="content"> PANTIDE can be a single toxic peptide of the three toxins we chose, Hv1a, Sf1a, and OAIP respectively with the addition of Galanthus nivalis agglutinin (GNA), for GNA has the function of acting as a carrier of PANTIDE from insect gut to the haemolymph and finally PANTIDE can transmit to the action site. </p>
 +
 +
<!----------Hv1a lectin----------->
 +
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974021" class="biobrick"><p class="content-1">BBa_K1974021</p></a>
 +
 +
<div>
 +
      <img src="https://static.igem.org/mediawiki/2016/b/b6/NCTU_FORMOSA_HL.png" class="picture">
 
</div>
 
</div>
  
−
<!-------cjBlue-LVA1------>
+
<p class="content">
−
<a href="http://parts.igem.org/Part:BBa_K2383001" class="biobrick"><p class="content-1">BBa_K2383001</p></a>
+
T7 promoter+RBS+Hv1a+linker+lectin+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for Hv1a and the snowdrop lectin linked with His-tag.
  
−
<img src="https://static.igem.org/mediawiki/2017/5/5e/Cjblue%2Blva.jpeg" class="bigphoto" width=70%;>
+
</p>
  
−
<p class="content">This chromoprotein from the ''Cnidopus japonicus'' sea anemone, cjBlue, naturally exhibits dark green color when expressed. Compared to some other chromoproteins, such as amilCP (<partinfo>BBa_K592009</partinfo>), amilGFP (<partinfo>BBa_K592010</partinfo>), spisPink (<partinfo>BBa_K1033932</partinfo>), asPink (<partinfo>BBa_K1033933</partinfo>) and aeBlue (<partinfo>BBa_K864401</partinfo>), the color development is slower. The color is readily observed in both LB or on agar plates after 24-48 hours of incubation. The sequence is codon optimized for expression in ''E coli''. We add a LVA tag after it to accelerate its degradation rate.</p>
+
<!---------Sf1a lectin---------->
 +
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974022" class="biobrick"><p class="content-1">BBa_K1974022</p></a>
  
−
<!-------cjBlue-LVA2------>
+
<div>
−
<a href="http://parts.igem.org/Part:BBa_K2383001" class="biobrick"><p class="content-1">BBa_K2383001</p></a>
+
      <img src="https://static.igem.org/mediawiki/2016/c/c3/NCTU_FORMOSA_SL.png" class="picture">
 +
</div>
  
−
<img src="https://static.igem.org/mediawiki/2017/5/5e/Cjblue%2Blva.jpeg" class="bigphoto" width=70%;>
+
<p class="content">
 +
T7 promoter+RBS+Sf1a+linker+lectin+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for Sf1a and the snowdrop lectin linked with His-tag.
  
−
<p class="content">This chromoprotein from the ''Cnidopus japonicus'' sea anemone, cjBlue, naturally exhibits dark green color when expressed. Compared to some other chromoproteins, such as amilCP (<partinfo>BBa_K592009</partinfo>), amilGFP (<partinfo>BBa_K592010</partinfo>), spisPink (<partinfo>BBa_K1033932</partinfo>), asPink (<partinfo>BBa_K1033933</partinfo>) and aeBlue (<partinfo>BBa_K864401</partinfo>), the color development is slower. The color is readily observed in both LB or on agar plates after 24-48 hours of incubation. The sequence is codon optimized for expression in ''E coli''. We add a LVA tag after it to accelerate its degradation rate.</p>
+
</p>
  
−
<!-------cjBlue-LVA3------>
+
<!--------OAIP lectin------>
−
<a href="http://parts.igem.org/Part:BBa_K2383001" class="biobrick"><p class="content-1">BBa_K2383001</p></a>
+
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974023" class="biobrick"><p class="content-1">BBa_K1974023</p></a>
  
−
<img src="https://static.igem.org/mediawiki/2017/5/5e/Cjblue%2Blva.jpeg" class="bigphoto" width=70%;>
+
<div>
 +
      <img src="https://static.igem.org/mediawiki/2016/2/27/NCTU_FORMOSA_OL.png" class="picture">
 +
</div>
  
−
<p class="content">This chromoprotein from the ''Cnidopus japonicus'' sea anemone, cjBlue, naturally exhibits dark green color when expressed. Compared to some other chromoproteins, such as amilCP (<partinfo>BBa_K592009</partinfo>), amilGFP (<partinfo>BBa_K592010</partinfo>), spisPink (<partinfo>BBa_K1033932</partinfo>), asPink (<partinfo>BBa_K1033933</partinfo>) and aeBlue (<partinfo>BBa_K864401</partinfo>), the color development is slower. The color is readily observed in both LB or on agar plates after 24-48 hours of incubation. The sequence is codon optimized for expression in ''E coli''. We add a LVA tag after it to accelerate its degradation rate.</p>
+
<p class="content">
 +
T7 promoter+RBS+OAIP+linker+lectin+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for OAIP and the snowdrop lectin linked with His-tag.
  
 +
</p>
 +
 +
<!---------------GS linker----------------->
 +
 +
<p class="title">PANTIDE with GS Linker</p>
 +
<p class="content"> In this part, to facilitate the expression of PANTIDE, we extended the linker with the substitution of GS linker that is even better than the original linker for the more correcter folding of the proteins structure.</p>
 +
 +
<!-----------OAIP GS linker--------------->
 +
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1974033" class="biobrick"><p class="content-1">BBa_K1974033</p></a>
 +
 +
<div>
 +
      <img src="https://static.igem.org/mediawiki/2016/d/d6/NCTU_FORMOSA_GS1.png" class="picture">
 +
</div>
 +
 +
<p class="content">
 +
T7 promoter+RBS+Hv1a+GS linker+lectin+linker+His-tag+terminator
 +
<br>PANTIDE that contains the gene coding for Hv1a and the snowdrop lectin. And improve the correct folding of PANTIDE by substituting with GS linker linked with His-tag.
 +
 +
 +
</p>
 +
 +
 +
 +
</div>
 
</section>
 
</section>
−
</h1>
+
 
 +
 
 +
 
 +
 
  
  

Revision as of 07:40, 26 October 2017

Composite Parts

We have known the function of the three toxic peptides of PANTIDE, Omega-hexatoxin-Hv1a, µ-segestritoxin-Sf1a, and Orally Active Insecticidal Peptide(OAIP) respectively and they are easy to express with the help of the bacteria E. coli BL21 Rosetta Gami strain. So, we then start to create BioBricks of these three toxins which we have obtained the nucleotide sequences. We use a strong T7 promoter that is suitable with the strain of E. coli we chose and also powerful at expressing proteins with multiple disulfide bonds, and we add His-tag for protein purification.

PANTIDE without Lectin

PANTIDE can be a single toxic peptide of the three toxins we chose, Hv1a, Sf1a, and OAIP respectively, which derive from the natural predators spiders.

BBa_K1974011

T7 promoter+RBS+Hv1a+linker+His-tag+terminator
PANTIDE that contains the gene coding for Hv1a linked with His-tag.

BBa_K1974012

T7 promoter+RBS+Sf1a+linker+His-tag+terminator
PANTIDE that contains the gene coding for Sf1a linked with His-tag.

BBa_K1974013

T7 promoter+RBS+OAIP+linker+His-tag+terminator
PANTIDE that contains the gene coding for OAIP linked with His-tag.

PANTIDE with Lectin

PANTIDE can be a single toxic peptide of the three toxins we chose, Hv1a, Sf1a, and OAIP respectively with the addition of Galanthus nivalis agglutinin (GNA), for GNA has the function of acting as a carrier of PANTIDE from insect gut to the haemolymph and finally PANTIDE can transmit to the action site.

BBa_K1974021

T7 promoter+RBS+Hv1a+linker+lectin+linker+His-tag+terminator
PANTIDE that contains the gene coding for Hv1a and the snowdrop lectin linked with His-tag.

BBa_K1974022

T7 promoter+RBS+Sf1a+linker+lectin+linker+His-tag+terminator
PANTIDE that contains the gene coding for Sf1a and the snowdrop lectin linked with His-tag.

BBa_K1974023

T7 promoter+RBS+OAIP+linker+lectin+linker+His-tag+terminator
PANTIDE that contains the gene coding for OAIP and the snowdrop lectin linked with His-tag.

PANTIDE with GS Linker

In this part, to facilitate the expression of PANTIDE, we extended the linker with the substitution of GS linker that is even better than the original linker for the more correcter folding of the proteins structure.

BBa_K1974033

T7 promoter+RBS+Hv1a+GS linker+lectin+linker+His-tag+terminator
PANTIDE that contains the gene coding for Hv1a and the snowdrop lectin. And improve the correct folding of PANTIDE by substituting with GS linker linked with His-tag.