Difference between revisions of "Team:Tec-Chihuahua/Parts"

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<html>
  
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<style>
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<h1>Parts</h1>
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.giro {
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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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  animation-name: spinner;  
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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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  animation-iteration-count: infinite;
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  -webkit-transform-origin: center center;
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@-webkit-keyframes spinner {
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<div class="highlight decoration_background">
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    from {
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<h3>Note</h3>
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  -webkit-transform: rotateX(0deg);
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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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to {
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  -moz-transform: rotateX(-360deg);
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  -ms-transform: rotateX(-360deg);
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  transform: rotateX(-360deg);
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}
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}
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</style>
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    <div class="parts8-bg"><br><br>
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<center><img src="https://static.igem.org/mediawiki/2018/c/c9/T--Tec-Chihuahua--Partsletters.png" width="13%" height="4%"/></center>
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    </div>
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        <div class="content">  
 +
        <div class="container">
  
  
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<groupparts>
 +
  <h3><b>Basic Parts Used</b></h3><br>
  
 +
  <b>Promoter T7 and RBS <a href="http://parts.igem.org/Part:BBa_K525998">[BBa_K525998]</a></b><br>
 +
      <p align="justify"> This unregulated T7 promoter, which has an integrated RBS, has high levels of transcription when the T7 RNA polymerase is present; thus, in order to induce expression of this BioBrick™ under the control IPTG will be used.
 +
      </p><br>
  
 +
  <b>PelB leader sequence<a href="http://parts.igem.org/Part:BBa_J32015"> [BBa_J32015]</a></b>
 +
      <p align="justify">The pelB leader sequence is a sequence of amino acids which, when attached to a protein, directs the protein to the bacterial periplasm. Protein secretion can increase the stability of cloned gene products. It was shown that the half-life of the recombinant proinsulin is increased 10-fold when the protein is secreted to the periplasmic space.
 +
      </p><br>
  
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<div class="column two_thirds_size">
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  <b>Defensin 1 <a href="http://parts.igem.org/Part:BBa_K1104301">[BBa_K1104301]</a></b>
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<div class="highlight decoration_B_full">
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      <p align="justify">Defensin 1 is synthesized in salivary glands and it plays an important role in the social immunity system of the bee, meaning that it is commonly passed through generations.
 +
Defensins unpolarize inner membranes by causing an ionic outer flow using preexistent channels. Apart from these processes, they are responsible for respiratory inhibition 3 minutes after they are added by directly attacking the respiration chain components.
 +
      </p><br>
  
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<h3>Adding parts to the registry</h3>
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  <b> Abaecin <a href="http://parts.igem.org/Part:BBa_K1104300">[BBa_K1104300]</a></b>
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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 align="justify"> Antimicrobial peptide that shows activity against both Gram-negative and positive bacteria. When being alone, it shows no activity against E. coli, at least at concentrations up to 200 μM. Hymenoptaecin a native peptide showed activity starting at 2μM concentration. When working together, helps by opening channels for the abaecin, which then gets to attack the DnaK, getting to interfere with a proper folding of the proteins.
 +
      </p><br>
  
−
<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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  <b>T1 from <i>E. coli</i> rrnB <a href="http://parts.igem.org/Part:BBa_B0010">[BBa_B0010]</a></b>
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<div class="button_link">
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      <p align="justify">A transcriptional terminator consisting of a 64 bp stem-loop. For the construction of our genetic circuitry, an efficient and reliable terminator was needed.
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<a href="http://parts.igem.org/Add_a_Part_to_the_Registry">
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      </p><br>
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ADD PARTS
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</a>
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</div>
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</div>
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<b><h3>New Additions to the Parts Registry</h3></b><br>
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</div>
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 +
<b>Modified 6xHis-tag <a href=" http://parts.igem.org/Part:BBa_K2834002">[BBa_K2834002]</a></b>
 +
<p align="justify"> A silently mutated amino acid motif in proteins that consists of  six histidine (His) residues, often at the N- or C-terminus of the protein. Polyhistidine-tags are often used for affinity purification of polyhistidine-tagged recombinant proteins expressed in Escherichia coli  and other prokaryotic expression systems.</p><br>
  
 +
<b>Apidaecin <a href="http://parts.igem.org/Part:BBa_K2834000">[BBa_K2834000]</a></b>
 +
<p align="justify"> It affects mainly Gram-negative bacteria, killing it impressively fast. Its mechanism of action is pretty unique. It has a total lack of pore-forming activity and its efficiency depends on the enantiomer used, meaning it has stereospecificity. Apidaecin gets to the ribosome 70s and the DnaK protein, leading to a protein synthesis inhibition when getting in contact with the ribosome, however, DnaK is the final target. This DnaK has an essential role in the folding of the proteins and in the DNA replication process, especially in the lambda phage.</p>
 +
<p id="validated"></p><br>
  
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<div class="column third_size">
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<b><h3>Composite Parts in our Creation</h3></b><br>
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<div class="highlight decoration_A_full">
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<h3>Inspiration</h3>
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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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<ul>
 
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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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<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Parts">2014 Tokyo Tech</a></li>
 
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</ul>
 
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</div>
 
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</div>
 
  
 +
  <b><p align="center">(T7 promoter + RBS) + (PelB) + (Defensin1) + (6x His-Tag) + (T1 terminator)</p> 
 +
  <center><a href="http://parts.igem.org/Part:BBa_K2834005">[BBa_K2834005]</a></b></center><br>
 +
  <img class="giro" src="https://static.igem.org/mediawiki/2018/a/ae/T--Tec-Chihuahua--A.png"/>
 +
  <i>(BBa_K525998) + (BBa_J32015) + (BBa_K1104301) + (BBa_K2834002) + (BBa_B0010)
 +
</i><br>
 +
  <p align="justify">This BioBrick™ contains the necessary genetic circuitry to be induced by IPTG and express the Defensin 1 peptide which will be transferred to the periplasmic region by the pelB. Post sonication this peptide can be isolated with the 6x His-Tag using an antiHis column.</p><br>
  
−
<div class="clear extra_space"></div>
 
  
 +
    <p align="center"><b>(T7 promoter + RBS) + (PelB) + (Abaecin) + (6x His-Tag) + (T1 terminator)</p> 
 +
    <center><a href="http://parts.igem.org/Part:BBa_K2834006">[BBa_K2834006]</a></b></center><br>
 +
    <img class="giro" src="https://static.igem.org/mediawiki/2018/9/91/T--Tec-Chihuahua--B.png"/>
 +
    <i>(BBa_K525998) + (BBa_J32015) + (BBa_K1104300) + (BBa_K2834002) + (BBa_B0010)</i><br>
 +
    <p align="justify">This BioBrick™ contains the necessary genetic circuitry to be induced by IPTG and express the Abaecin peptide which will be transferred to the periplasmic region by the pelB. Post sonication this peptide can be isolated with the 6x His-Tag using a antiHis column.</p><br>
  
 +
    <p align="center"><b>(T7 promoter + RBS) + (PelB) + (Apidaecin) + (6x His-Tag) + (T1 terminator)</b></p>
 +
    <center><b><a href="http://parts.igem.org/Part:BBa_K2834003">[BBa_K2834003]</a></b></center><br>
 +
    <img class="giro" src="https://static.igem.org/mediawiki/2018/0/03/T--Tec-Chihuahua--D.png" width="100%" height=80%"/><br>
 +
    <i>(BBa_K525998) + (BBa_J32015) + (BBa_K2834000) + (BBa_K2834002) + (BBa_B0010)</i><br>
 +
    <p align="justify">This BioBrick™ contains the necessary genetic circuitry to be induced by IPTG and express the Apidaecin peptide which will be transferred to the periplasmic region by the pelB. Post sonication this peptide can be isolated with the 6x His-Tag using a antiHis column.</p><br>
  
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  <div class="container">
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<div class="column full_size">
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      <button class="collapsible"><h4 align="center"><b>References</b></h4></button>
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      <div class="contentk">
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<h3>What information do I need to start putting my parts on the Registry?</h3>
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  <ol><br>
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<p>The information needed to initially create a part on the Registry is:</p>
+
      <li>Ilyasov, R. A., Gaifullina, L. R., Saltykova, E. S., Poskryakov, A. V., & Nikolaenko, A. G. (2013). Defensins in the honeybee antiinfectious protection. Journal of Evolutionary Biochemistry and Physiology, 49(1), 1–9. https://doi.org/10.1134/s0022093013010015</li>
−
<ul>
+
      <li>Cociancich, S., Ghazi, A., Hetru, C., Hoffman, J. A., & Letellier, L. (1993). Insect Defensin, an Inducible Antibacterial Peptide, Forms Voltage-dependent Channels in Micrococcus luteus. The Journal of Biological Chemistry, 268(26), 19239-19245
−
<li>Part Name</li>
+
      <li>Shen, X., Ye, G., Cheng, X., Yu, C., Hu, C., Allosaar, I. (2010). Characterization of an abaecin-like antimicrobial peptide identified from a Pteromalus puparum cDNA clone. Retrieved from: https://www.sciencedirect.com/science/article/pii/S0022201110001114</li>
−
<li>Part type</li>
+
      <li>Castle, M., Nazarian, A., Yi, S., Tempst, P. (1999). Lethal effects of apidaecin on Escherichia coli involve sequential molecular interactions with diverse targets. Retrieved from: http://www.jbc.org/content/274/46/32555.full.pdf</li>
−
<li>Creator</li>
+
      <li>Cociancich, S., Ghazi, A., Hetru, C., Hoffman, J. A., & Letellier, L. (1993). Insect Defensin, an Inducible Antibacterial Peptide, Forms Voltage-dependent Channels in Micrococcus luteus. The Journal of Biological Chemistry, 268(26), 19239-19245</li>
−
<li>Sequence</li>
+
  </ol>
−
<li>Short Description (60 characters on what the DNA does)</li>
+
<br>
−
<li>Long Description (Longer description of what the DNA does)</li>
+
  </div>
−
<li>Design considerations</li>
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</ul>
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+
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<p>
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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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<style>
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.collapsible {
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    background-color: #777;
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    cursor: pointer;
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    width: 100%;
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    border: none;
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    text-align: left;
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    outline: none;
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    font-size: 15px;
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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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<groupparts>iGEM18 Tec-Chihuahua</groupparts>
 
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{{:Team:Tec-Chihuahua/Templates/footer}}

Latest revision as of 23:15, 17 October 2018

Erwinions



Basic Parts Used


Promoter T7 and RBS [BBa_K525998]

This unregulated T7 promoter, which has an integrated RBS, has high levels of transcription when the T7 RNA polymerase is present; thus, in order to induce expression of this BioBrick™ under the control IPTG will be used.


PelB leader sequence [BBa_J32015]

The pelB leader sequence is a sequence of amino acids which, when attached to a protein, directs the protein to the bacterial periplasm. Protein secretion can increase the stability of cloned gene products. It was shown that the half-life of the recombinant proinsulin is increased 10-fold when the protein is secreted to the periplasmic space.


Defensin 1 [BBa_K1104301]

Defensin 1 is synthesized in salivary glands and it plays an important role in the social immunity system of the bee, meaning that it is commonly passed through generations. Defensins unpolarize inner membranes by causing an ionic outer flow using preexistent channels. Apart from these processes, they are responsible for respiratory inhibition 3 minutes after they are added by directly attacking the respiration chain components.


Abaecin [BBa_K1104300]

Antimicrobial peptide that shows activity against both Gram-negative and positive bacteria. When being alone, it shows no activity against E. coli, at least at concentrations up to 200 μM. Hymenoptaecin a native peptide showed activity starting at 2μM concentration. When working together, helps by opening channels for the abaecin, which then gets to attack the DnaK, getting to interfere with a proper folding of the proteins.


T1 from E. coli rrnB [BBa_B0010]

A transcriptional terminator consisting of a 64 bp stem-loop. For the construction of our genetic circuitry, an efficient and reliable terminator was needed.


New Additions to the Parts Registry


Modified 6xHis-tag [BBa_K2834002]

A silently mutated amino acid motif in proteins that consists of six histidine (His) residues, often at the N- or C-terminus of the protein. Polyhistidine-tags are often used for affinity purification of polyhistidine-tagged recombinant proteins expressed in Escherichia coli and other prokaryotic expression systems.


Apidaecin [BBa_K2834000]

It affects mainly Gram-negative bacteria, killing it impressively fast. Its mechanism of action is pretty unique. It has a total lack of pore-forming activity and its efficiency depends on the enantiomer used, meaning it has stereospecificity. Apidaecin gets to the ribosome 70s and the DnaK protein, leading to a protein synthesis inhibition when getting in contact with the ribosome, however, DnaK is the final target. This DnaK has an essential role in the folding of the proteins and in the DNA replication process, especially in the lambda phage.


Composite Parts in our Creation


(T7 promoter + RBS) + (PelB) + (Defensin1) + (6x His-Tag) + (T1 terminator)

[BBa_K2834005]

(BBa_K525998) + (BBa_J32015) + (BBa_K1104301) + (BBa_K2834002) + (BBa_B0010)

This BioBrick™ contains the necessary genetic circuitry to be induced by IPTG and express the Defensin 1 peptide which will be transferred to the periplasmic region by the pelB. Post sonication this peptide can be isolated with the 6x His-Tag using an antiHis column.


(T7 promoter + RBS) + (PelB) + (Abaecin) + (6x His-Tag) + (T1 terminator)

[BBa_K2834006]

(BBa_K525998) + (BBa_J32015) + (BBa_K1104300) + (BBa_K2834002) + (BBa_B0010)

This BioBrick™ contains the necessary genetic circuitry to be induced by IPTG and express the Abaecin peptide which will be transferred to the periplasmic region by the pelB. Post sonication this peptide can be isolated with the 6x His-Tag using a antiHis column.


(T7 promoter + RBS) + (PelB) + (Apidaecin) + (6x His-Tag) + (T1 terminator)

[BBa_K2834003]


(BBa_K525998) + (BBa_J32015) + (BBa_K2834000) + (BBa_K2834002) + (BBa_B0010)

This BioBrick™ contains the necessary genetic circuitry to be induced by IPTG and express the Apidaecin peptide which will be transferred to the periplasmic region by the pelB. Post sonication this peptide can be isolated with the 6x His-Tag using a antiHis column.



  1. Ilyasov, R. A., Gaifullina, L. R., Saltykova, E. S., Poskryakov, A. V., & Nikolaenko, A. G. (2013). Defensins in the honeybee antiinfectious protection. Journal of Evolutionary Biochemistry and Physiology, 49(1), 1–9. https://doi.org/10.1134/s0022093013010015
  2. Cociancich, S., Ghazi, A., Hetru, C., Hoffman, J. A., & Letellier, L. (1993). Insect Defensin, an Inducible Antibacterial Peptide, Forms Voltage-dependent Channels in Micrococcus luteus. The Journal of Biological Chemistry, 268(26), 19239-19245
  3. Shen, X., Ye, G., Cheng, X., Yu, C., Hu, C., Allosaar, I. (2010). Characterization of an abaecin-like antimicrobial peptide identified from a Pteromalus puparum cDNA clone. Retrieved from: https://www.sciencedirect.com/science/article/pii/S0022201110001114
  4. Castle, M., Nazarian, A., Yi, S., Tempst, P. (1999). Lethal effects of apidaecin on Escherichia coli involve sequential molecular interactions with diverse targets. Retrieved from: http://www.jbc.org/content/274/46/32555.full.pdf
  5. Cociancich, S., Ghazi, A., Hetru, C., Hoffman, J. A., & Letellier, L. (1993). Insect Defensin, an Inducible Antibacterial Peptide, Forms Voltage-dependent Channels in Micrococcus luteus. The Journal of Biological Chemistry, 268(26), 19239-19245


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