Team:Aberdeen Scotland/Project/Design

From 2014.igem.org

(Difference between revisions)
(Created page with "<html> <head> <!-- Charset --> <meta charset="UTF-8"> <title>Team:Aberdeen Scotland/Project/Design - 2014.ogem.org</title> <!-- JavaScript --> <script src="http://2014.i...")
Line 61: Line 61:
<!-- SECTION HEAD -->
<!-- SECTION HEAD -->
<div class="t_overview">
<div class="t_overview">
-
<h1><i>E. coli</i>-based Trypanosomiasis Diagnostic System</h1>
+
<h1>Our System</h1>
<p>Getting back at the Sleeping sickness by detecting it early.</p>
<p>Getting back at the Sleeping sickness by detecting it early.</p>
</div> <br class="clear"> <!-- END OF HEAD -->
</div> <br class="clear"> <!-- END OF HEAD -->
Line 67: Line 67:
<!-- PAGE CONTENT -->
<!-- PAGE CONTENT -->
<div class="main_content">
<div class="main_content">
-
<p>The goal of our project is to to develop a novel method for diagnosing Trypanosomiasis. Our aim is to provide a simpler, cheaper alternative to current methods that would be more versatile in developing countries and their remote regions. We wish to create a test that would be portable, endure harsh environmental conditions and most importantly be sensitive to the early stages of the disease.</p>
+
<p>We aim to design an E. coli-based detection system that can be used for the diagnosis of trypanosomiasis using patient serum polyclonal antibodies.</p>
 +
<p>We will create Antigen-43 and Ice Nucleation Protein constructs, to be expressed on surface of E. coli, that carry an additional sequence of amino acids referred to as a ‘mimotope’. A mimotope (mimic epitope), functions as a target for disease-specific antibodies in the blood, which may be present due to an active immune response to a pathogen. A green fluorescence response indicates a positive disease diagnosis.</p>
 +
<img src="https://static.igem.org/mediawiki/2014/c/c0/Design_diag.PNG">
 +
<p>Antigen-43 (Ag43) and Ice Nucleation Protein (INP), are autotransporter proteins that exist endogenously on the surface of E. coli; they can transport large peptides for cell-surface display. We have concentrated on creating ‘BioBrick’ versions of these for submission to the iGEM 2014 repository.</p>
 +
</p>The Ag43 has had its PstI restriction sites removed to make it BioBrick compatible, the two Beta-hairpins removed to prevent cellular auto-aggregation. A BglII/HindIII restriction site has been added for easy future insertions of desired sequences. We have also introduced a FLAG-tag to create a demonstrable ‘proof of concept’ for antibody aggregation-based detection. We intend to also create a His-tag version.</p>
 +
<p>We began with an INP-YFP (Yellow Fluorescent Protein) BioBrick created by a previous team. To this we added a BglII/HindIII restriction site. We also introduced a FLAG-tag and His-tag to this site in 2 separate versions.</p>
</div> <br class="clear"> <!-- END OF PAGE CONTENT -->
</div> <br class="clear"> <!-- END OF PAGE CONTENT -->
</div> <!-- END OF CONTAINER -->
</div> <!-- END OF CONTAINER -->

Revision as of 23:05, 14 August 2014

Team:Aberdeen Scotland/Project/Design - 2014.ogem.org



Our System

Getting back at the Sleeping sickness by detecting it early.


We aim to design an E. coli-based detection system that can be used for the diagnosis of trypanosomiasis using patient serum polyclonal antibodies.

We will create Antigen-43 and Ice Nucleation Protein constructs, to be expressed on surface of E. coli, that carry an additional sequence of amino acids referred to as a ‘mimotope’. A mimotope (mimic epitope), functions as a target for disease-specific antibodies in the blood, which may be present due to an active immune response to a pathogen. A green fluorescence response indicates a positive disease diagnosis.

Antigen-43 (Ag43) and Ice Nucleation Protein (INP), are autotransporter proteins that exist endogenously on the surface of E. coli; they can transport large peptides for cell-surface display. We have concentrated on creating ‘BioBrick’ versions of these for submission to the iGEM 2014 repository.

The Ag43 has had its PstI restriction sites removed to make it BioBrick compatible, the two Beta-hairpins removed to prevent cellular auto-aggregation. A BglII/HindIII restriction site has been added for easy future insertions of desired sequences. We have also introduced a FLAG-tag to create a demonstrable ‘proof of concept’ for antibody aggregation-based detection. We intend to also create a His-tag version.

We began with an INP-YFP (Yellow Fluorescent Protein) BioBrick created by a previous team. To this we added a BglII/HindIII restriction site. We also introduced a FLAG-tag and His-tag to this site in 2 separate versions.