Difference between revisions of "Team:NCTU Formosa/Composite Part"

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<div class="p02">
 
<div class="p02">
 
<div class="content"></div>
 
<div class="content"></div>
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<h2>Composite Part</h2>
  
 +
<table>
 +
  <tr><td width="20%"></td><td width="13%"></td><td width="67%"></td></tr>
 +
 +
  <tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694013">BBa_K1694013</a></span><br>
 +
                                  OmpA-anti-VEGF<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694014">BBa_K1694014</a></span><br>
 +
                                OmpA-anti-EGFR<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694015">BBa_K1694015</a></span><br>
 +
                                OmpA-anti-HER2<br>
 +
</td><td></td><td>In order to change the scFv parts easily, we added a NcoI restriction site between OmpA and scFv so that we can <B>change various scFv DNA sequence</B> using the NcoI restriction enzyme.</td></tr>
 +
 +
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694023">BBa_K1694023</a></span><br>
 +
                                  Pcons+RBS+OmpA-anti-VEGF<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694024">BBa_K1694024</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-EGFR<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694025">BBa_K1694025</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-HER2<br>
 +
</td><td></td><td>By ligating the constitutive promoter (BBa_J23101), strong ribosome binding site (BBa_B0034) and Lpp-OmpA-scFv, we were able to display scFv on the <i>E.coli</i> outer membrane continuously.
 +
Having this part, we can co-transform with other parts in order to produce color as the detection signal.<br><br>
 +
 +
In addition, by co-transforming these different types of <i>E.coli</i> with different fluorescence or color as signals, we are able to create a platform which can detect <B>multimarker</B> and consequently achieve combination therapy.
 +
</td></tr>
 +
 +
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694033">BBa_K1694033</a></span><br>
 +
                                  Pcons+RBS+OmpA-anti-VEGF+RBS+GFP+Ter<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694034">BBa_K1694034</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-EGFR+RBS+GFP+Ter<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694035">BBa_K1694035</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-HER2+RBS+GFP+Ter<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694044">BBa_K1694044</a></span><br>
 +
                      Pcons+RBS+OmpA-anti-EGFR+RBS+RFP+Ter<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694045">BBa_K1694045</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-HER2+RBS+BFP+Ter<br>
 +
                               
 +
</td><td></td><td>The most commonly constructing way of composite parts is to ligate the required parts together. We also provide this kind of parts.<br>
 +
At the back of the Lpp-OmpA-scFv part, we ligated the weaker ribosome biding site (BBa_B0030), different fluorescent protein and terminator (BBa_J61048) to make it continuously and simultaneously express the fluorescence and the scFv . We used a weak ribosome binding site to ensure that scFv's production will not be affected.
 +
</td></tr>
 +
 +
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694053">BBa_K1694053</a></span><br>
 +
                                  Pcons+RBS+OmpA-anti-VEGF+RBS+amilCP+Ter<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694054">BBa_K1694054</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-EGFR+RBS+amilCP+Ter<br>
 +
                      <span><a href="http://parts.igem.org/Part:BBa_K1694055">BBa_K1694035</a></span><br>
 +
                                Pcons+RBS+OmpA-anti-HER2+RBS+amilCP+Ter<br>
 +
</td><td></td><td>Chromoprotein is another example of what can be added when making your own probe.<br>
 +
We constructed this part as the Pcons+RBS+OmpA-scFv+RBS+fluorescent protein+Terminator part.
 +
</td></tr>
 +
 +
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694027">BBa_K1694027</a></span><br>
 +
                                  Pinduce+RBS+FadL-GBP<br>
 +
</td><td></td><td>By ligating the induced promoter (BBa_R0010), ribosome binding site (BBa_B0034), FadL-GBP and terminator (BBa_J61048), we can continuously display the GBP on the <i>E.coli</i> outer membrane.<br>
 +
Then we co-transform the OmpA-scFv parts and this FadL-GBP parts into the same <i>E.coli</i>, hence allowing our <i>E.coli</i> to bind on gold chips and detect antigens simultaneously.
 +
</td></tr>
 +
 +
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694037">BBa_K1694037</a></span><br>
 +
                                Pinduce+RBS+FadL-GBP+RBS+GFP+Ter<br>
 +
</td><td></td><td>With this part, we can testt the GBP function by observing the green fluorescence on the gold chip.
 +
</td></tr>
 +
 +
</table>
 +
</div>
 +
 +
<div class="content">
 +
<div class="reference">
 +
<b>Reference<br></b>
 +
[1] C Hartmann et al. (2010) Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce a functionally equivalent anti-EGFR immune response http://www.nature.com/onc/journal/v29/n32/pdf/onc2010195a.pdf<br>
 +
[2] DrugBank: Bevacizumab (DB00112) http://www.drugbank.ca/drugs/DB00112<br>
 +
[3] DrugBank: Cetuximab (DB00002) http://www.drugbank.ca/drugs/DB00002<br>
 +
[4] DrugBank: Trastuzumab (DB00072) http://www.drugbank.ca/drugs/DB00072<br>
 +
[5] Tae Jung Park et al. (2009) Development of a whole-cell biosensor by cell surface display of a gold-binding polypeptide on the gold surface<br>
 +
</div>
 +
</div>
  
  

Revision as of 17:43, 16 September 2015

Composite Parts

Composite Part

BBa_K1694013
OmpA-anti-VEGF
BBa_K1694014
OmpA-anti-EGFR
BBa_K1694015
OmpA-anti-HER2
In order to change the scFv parts easily, we added a NcoI restriction site between OmpA and scFv so that we can change various scFv DNA sequence using the NcoI restriction enzyme.
BBa_K1694023
Pcons+RBS+OmpA-anti-VEGF
BBa_K1694024
Pcons+RBS+OmpA-anti-EGFR
BBa_K1694025
Pcons+RBS+OmpA-anti-HER2
By ligating the constitutive promoter (BBa_J23101), strong ribosome binding site (BBa_B0034) and Lpp-OmpA-scFv, we were able to display scFv on the E.coli outer membrane continuously. Having this part, we can co-transform with other parts in order to produce color as the detection signal.

In addition, by co-transforming these different types of E.coli with different fluorescence or color as signals, we are able to create a platform which can detect multimarker and consequently achieve combination therapy.
BBa_K1694033
Pcons+RBS+OmpA-anti-VEGF+RBS+GFP+Ter
BBa_K1694034
Pcons+RBS+OmpA-anti-EGFR+RBS+GFP+Ter
BBa_K1694035
Pcons+RBS+OmpA-anti-HER2+RBS+GFP+Ter
BBa_K1694044
Pcons+RBS+OmpA-anti-EGFR+RBS+RFP+Ter
BBa_K1694045
Pcons+RBS+OmpA-anti-HER2+RBS+BFP+Ter
The most commonly constructing way of composite parts is to ligate the required parts together. We also provide this kind of parts.
At the back of the Lpp-OmpA-scFv part, we ligated the weaker ribosome biding site (BBa_B0030), different fluorescent protein and terminator (BBa_J61048) to make it continuously and simultaneously express the fluorescence and the scFv . We used a weak ribosome binding site to ensure that scFv's production will not be affected.
BBa_K1694053
Pcons+RBS+OmpA-anti-VEGF+RBS+amilCP+Ter
BBa_K1694054
Pcons+RBS+OmpA-anti-EGFR+RBS+amilCP+Ter
BBa_K1694035
Pcons+RBS+OmpA-anti-HER2+RBS+amilCP+Ter
Chromoprotein is another example of what can be added when making your own probe.
We constructed this part as the Pcons+RBS+OmpA-scFv+RBS+fluorescent protein+Terminator part.
BBa_K1694027
Pinduce+RBS+FadL-GBP
By ligating the induced promoter (BBa_R0010), ribosome binding site (BBa_B0034), FadL-GBP and terminator (BBa_J61048), we can continuously display the GBP on the E.coli outer membrane.
Then we co-transform the OmpA-scFv parts and this FadL-GBP parts into the same E.coli, hence allowing our E.coli to bind on gold chips and detect antigens simultaneously.
BBa_K1694037
Pinduce+RBS+FadL-GBP+RBS+GFP+Ter
With this part, we can testt the GBP function by observing the green fluorescence on the gold chip.
Reference
 [1] C Hartmann et al. (2010) Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce a functionally equivalent anti-EGFR immune response http://www.nature.com/onc/journal/v29/n32/pdf/onc2010195a.pdf
[2] DrugBank: Bevacizumab (DB00112) http://www.drugbank.ca/drugs/DB00112
[3] DrugBank: Cetuximab (DB00002) http://www.drugbank.ca/drugs/DB00002
[4] DrugBank: Trastuzumab (DB00072) http://www.drugbank.ca/drugs/DB00072
[5] Tae Jung Park et al. (2009) Development of a whole-cell biosensor by cell surface display of a gold-binding polypeptide on the gold surface


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