Difference between revisions of "Team:TecCEM/Basic Part"

 
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         text-decoration: none;
 
         text-decoration: none;
 
     }
 
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     div.parts-div a:vistied {
 
     div.parts-div a:vistied {
 
         color: #333333 !important;
 
         color: #333333 !important;
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     }
 
     }
 
</style>
 
</style>
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<div id="gif-title" class="text-red">
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<div id="gif-title" class="text-parts">
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     <img src="https://static.igem.org/mediawiki/2018/a/aa/T--TecCEM--Cells.gif" alt="Cell Gif">
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     <img src="https://static.igem.org/mediawiki/2018/4/4e/T--TecCEM--Cells--Interlab.gif" alt="Cell Gif">
 
     <h1>Basic Parts</h1>
 
     <h1>Basic Parts</h1>
 
</div>
 
</div>
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                         GST and
 
                         GST and
 
                         the target protein (Harper & Speicher, 2013).</p>
 
                         the target protein (Harper & Speicher, 2013).</p>
−
 
+
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/4/4f/T--TecCEM--GST3DStructure.png" class="figure-img img-fluid rounded"
 +
                                alt="GST Coding Site">
 +
                            <figcaption class="figure-caption">Figure 1. GST 3D structure, modelled with Swiss-Model</figcaption>
 +
                        </figure>
 +
                    </div>
 
                     <h2 id="K2719001"><a href="http://parts.igem.org/Part:BBa_K2719001">BBa_K2719001</a>: Tenascin
 
                     <h2 id="K2719001"><a href="http://parts.igem.org/Part:BBa_K2719001">BBa_K2719001</a>: Tenascin
 
                         Domain V</h2>
 
                         Domain V</h2>
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                         been
 
                         been
 
                         reported that it participated stem cells signalling.</p>
 
                         reported that it participated stem cells signalling.</p>
−
 
+
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/3/30/T--TecCEM--TCD5DomainV3DStructure.png"
 +
                                class="figure-img img-fluid rounded" alt="Tenascin
 +
                                Domain V">
 +
                            <figcaption class="figure-caption">Figure 2. Tenascin 5 domain V 3D structure, modelled
 +
                                with Swiss-Model</figcaption>
 +
                        </figure>
 +
                    </div>
 
                     <h2 id="K2719002"><a href="http://parts.igem.org/Part:BBa_K2719002">BBa_K2719002</a>: GST-Tenascin
 
                     <h2 id="K2719002"><a href="http://parts.igem.org/Part:BBa_K2719002">BBa_K2719002</a>: GST-Tenascin
−
                         Domain V
+
                         Domain V Fusion Protein</h2>
−
                        Fusion
+
−
                        Protein</h2>
+
 
                     <p>This part is the fusion between both proteins and that helps us to simplify tenascin
 
                     <p>This part is the fusion between both proteins and that helps us to simplify tenascin
 
                         purification. GST
 
                         purification. GST
 
                         has tenascin fused on the N-terminal. For more information see BBa_K2719001 and BBa_K2719003.</p>
 
                         has tenascin fused on the N-terminal. For more information see BBa_K2719001 and BBa_K2719003.</p>
−
 
+
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/1/10/T--TecCEM--TCD5%2BGST3DStructure.png" class="figure-img img-fluid rounded"
 +
                                alt="GST-Tenascin Domain V Fusion Protein">
 +
                            <figcaption class="figure-caption">Figure 3. Tenascin V- GST fusion protein 3D structure,
 +
                                modeled with Swiss-Model</figcaption>
 +
                        </figure>
 +
                    </div>
 
                     <h2 id="K2719003"><a href="http://parts.igem.org/Part:BBa_K2719003">BBa_K2719003</a>: Optimized GST
 
                     <h2 id="K2719003"><a href="http://parts.igem.org/Part:BBa_K2719003">BBa_K2719003</a>: Optimized GST
 
                         Coding
 
                         Coding
 
                         site</h2>
 
                         site</h2>
−
                     <p>GST coding sequence (see part BBa_K2719000) with codon optimization for E.coli BL21 (DE3) to
+
                     <p>GST coding sequence (see part BBa_K2719000) with codon optimization for <i>E.coli</i> BL21 (DE3) to
 
                         avoid
 
                         avoid
 
                         complications in the production of this recombinant protein.</p>
 
                         complications in the production of this recombinant protein.</p>
−
 
+
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/4/4f/T--TecCEM--GST3DStructure.png" class="figure-img img-fluid rounded"
 +
                                alt="GST 3D structure, modelled with Swiss-Model">
 +
                            <figcaption class="figure-caption">Figure 4. GST 3D structure, modelled with Swiss-Model</figcaption>
 +
                        </figure>
 +
                    </div>
 
                     <h2 id="K2719004"><a href="http://parts.igem.org/Part:BBa_K2719004">BBa_K2719004</a>: Tenascin
 
                     <h2 id="K2719004"><a href="http://parts.igem.org/Part:BBa_K2719004">BBa_K2719004</a>: Tenascin
 
                         Fibronectin
 
                         Fibronectin
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                         is domain
 
                         is domain
 
                         V (see part BBa_K2719001).</p>
 
                         V (see part BBa_K2719001).</p>
−
 
+
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/e/e9/T--TecCEM--TCD53DStructure.png" class="figure-img img-fluid rounded"
 +
                                alt="Tenascin Fibronectin Domains">
 +
                            <figcaption class="figure-caption">Figura 5. Tenascin V Fibronectin Domains 3D Structure.
 +
                                Modelled with Swiss-Model</figcaption>
 +
                        </figure>
 +
                    </div>
 
                     <h2 id="K2719006"><a href="http://parts.igem.org/Part:BBa_K2719006">BBa_K2719006</a>: Collagen V-
 
                     <h2 id="K2719006"><a href="http://parts.igem.org/Part:BBa_K2719006">BBa_K2719006</a>: Collagen V-
 
                         like
 
                         like
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                         proline. To overcome this challenge, we designed this part to incorporate hydroxyproline, a
 
                         proline. To overcome this challenge, we designed this part to incorporate hydroxyproline, a
 
                         non-canonical amino acid. For its proper incorporation, this part is optimized for its use on
 
                         non-canonical amino acid. For its proper incorporation, this part is optimized for its use on
−
                         E.coli
+
                         <i>E.coli</i>
 
                         Rosetta (DE3).</p>
 
                         Rosetta (DE3).</p>
−
 
+
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/0/0e/T--TecCEM--ColV2DStructure.png" class="figure-img img-fluid rounded"
 +
                                alt="Collagen V-
 +
                                like
 +
                                protein coding
 +
                                site">
 +
                            <figcaption class="figure-caption">Figure 6. 2D structure of Collagen V-like</figcaption>
 +
                        </figure>
 +
                    </div>
 
                     <h2 id="K2719008"><a href="http://parts.igem.org/Part:BBa_K2719008">BBa_K2719008</a>: Leptin coding
 
                     <h2 id="K2719008"><a href="http://parts.igem.org/Part:BBa_K2719008">BBa_K2719008</a>: Leptin coding
 
                         site.</h2>
 
                         site.</h2>
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                         thus,
 
                         thus,
 
                         accelerating cell proliferation.</p>
 
                         accelerating cell proliferation.</p>
 +
                    <div class="text-center">
 +
                        <figure class="figure text-left">
 +
                            <img src="https://static.igem.org/mediawiki/2018/d/d3/T--TecCEM--Lep3DStructure.png" class="figure-img img-fluid rounded"
 +
                                alt="Leptin coding site">
 +
                            <figcaption class="figure-caption">Figure 7. Leptin 3D prediction modelled with Swiss Model</figcaption>
 +
                        </figure>
 +
                    </div>
 
                 </div>
 
                 </div>
 
             </div>
 
             </div>

Latest revision as of 00:44, 18 October 2018

Cell Gif

Basic Parts

Basic Parts

BBa_K2719000: GST Coding site

Glutathione S-transferase (GST) is a peptide derived from Schistosoma japonicum, in which a target protein may be fused on the N-terminal. This fusion allows the purification of the target protein using glutathione affinity and maximize the presence of the protein in soluble state, rather than on inclusion bodies. This tag may be later removed by adding a protease recognition site between GST and the target protein (Harper & Speicher, 2013).

GST Coding Site
Figure 1. GST 3D structure, modelled with Swiss-Model

BBa_K2719001: Tenascin Domain V

Tenascin is a human protein that is involved in tissue regeneration processes. The domain V contains heparin binding sites that are fundamental for our scaffold. Also contains multiple growth factor binding sites and, even when we are only working with leptin is important because it would help in the improvement of tissue regeneration. In addition, it has been reported that it participated stem cells signalling.

Tenascin Domain V
Figure 2. Tenascin 5 domain V 3D structure, modelled with Swiss-Model

BBa_K2719002: GST-Tenascin Domain V Fusion Protein

This part is the fusion between both proteins and that helps us to simplify tenascin purification. GST has tenascin fused on the N-terminal. For more information see BBa_K2719001 and BBa_K2719003.

GST-Tenascin Domain V Fusion Protein
Figure 3. Tenascin V- GST fusion protein 3D structure, modeled with Swiss-Model

BBa_K2719003: Optimized GST Coding site

GST coding sequence (see part BBa_K2719000) with codon optimization for E.coli BL21 (DE3) to avoid complications in the production of this recombinant protein.

GST 3D structure, modelled with Swiss-Model
Figure 4. GST 3D structure, modelled with Swiss-Model

BBa_K2719004: Tenascin Fibronectin Domains

Coding sequence from tenascin fibronectin domains I-V have the property of interacting with the proliferation of fibroblast, but the domain with the main characteristic previously mentioned is domain V (see part BBa_K2719001).

Tenascin Fibronectin Domains
Figura 5. Tenascin V Fibronectin Domains 3D Structure. Modelled with Swiss-Model

BBa_K2719006: Collagen V- like protein coding site

Collagen V is one of the main proteins involved in the extracellular matrix, thus it would be important in the structure of our scaffold. The main challenge for producing recombinant collagen V is that for its correct structure it requires post-traductional modifications, specially the hydroxylation of proline. To overcome this challenge, we designed this part to incorporate hydroxyproline, a non-canonical amino acid. For its proper incorporation, this part is optimized for its use on E.coli Rosetta (DE3).

Collagen V- like protein coding site
Figure 6. 2D structure of Collagen V-like

BBa_K2719008: Leptin coding site.

This is one of the most important parts because is the growth factor that we are using for accelerating the process of skin regeneration. Is a protein that is found in adipocytes cells on many living beings and it has been discovered that it has a regenerating property, because it has been found in many wound repair processes. It has been shown that it increases glucose intake speeding up metabolism, thus, accelerating cell proliferation.

Leptin coding site
Figure 7. Leptin 3D prediction modelled with Swiss Model