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

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<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694027">BBa_K1694027</a></span><br>
 
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694027">BBa_K1694027</a></span><br>
−
                                   P<sub>induce</sub>+RBS+FadL-GBP<br>  
+
                                   P<sub>cons</sub>+RBS+FadL-GBP<br>  
−
</td><td></td><td>By ligating the induced promoter (<a href="http://parts.igem.org/Part:BBa_R0010">BBa_R0010</a>), ribosome binding site (<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>) and FadL-GBP, we can continuously display the GBP on the <i>E.coli</i> outer membrane.<br>
+
</td><td></td><td>By ligating the induced promoter (<a href="http://parts.igem.org/Part:BBa_J23110">BBa_J23
 +
110</a>), ribosome binding site (<a href="http://parts.igem.org/Part:BBa_B0034">BBa_B0034</a>) and FadL-GBP, 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.  
 
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>  
 
</td></tr>  
  
 
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694037">BBa_K1694037</a></span><br>
 
<tr><td class="part"><span><a href="http://parts.igem.org/Part:BBa_K1694037">BBa_K1694037</a></span><br>
−
                                 P<sub>induce</sub>+RBS+FadL-GBP+RBS+GFP+Ter<br>  
+
                                 P<sub>cons</sub>+RBS+FadL-GBP+RBS+GFP+Ter<br>  
 
</td><td></td><td>With this part, we can test the GBP function by observing the green fluorescence on the gold chip. Terminator  is (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>),
 
</td><td></td><td>With this part, we can test the GBP function by observing the green fluorescence on the gold chip. Terminator  is (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>),
 
</td></tr>  
 
</td></tr>  

Revision as of 23:32, 18 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_K1694055
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
Pcons+RBS+FadL-GBP
By ligating the induced promoter (BBa_J23 110), ribosome binding site (BBa_B0034) and FadL-GBP, 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
Pcons+RBS+FadL-GBP+RBS+GFP+Ter
With this part, we can test the GBP function by observing the green fluorescence on the gold chip. Terminator is (BBa_B0015),
Reference
 [1] C Hartmann et al. (2010) Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce a functionally equivalent anti-EGFR immune response
[2] DrugBank: Bevacizumab (DB00112)
[3] DrugBank: Cetuximab (DB00002)
[4] DrugBank: Trastuzumab (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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