Difference between revisions of "Team:IISc-Bangalore"

(Updated abstract and title)
Line 12: Line 12:
 
<div class="column full_size" >
 
<div class="column full_size" >
 
<h1>iFLOAT</h1>
 
<h1>iFLOAT</h1>
−
<p>iFLOAT is our attempt to develop a novel protein purification strategy using cyanobacterial gas vesicle tagged proteins. Our project strives to reduce downstream recombinant protein purification costs and will vastly improve cost-effectiveness and productivity of industries reliant on recombinant protein production. </p>
+
<h2>a multifaceted approach to cluster bioengineered gas vesicles in vitro and enhance their flotation</h2>
  
−
<p>To facilitate this, we are engineering E. coli to express gas vesicle proteins (gvpA and gvpC) from cyanobacteria (Planktothrix rubescens). The recombinant protein of interest will be fused to gvpC using an amino acid linker comprising the sequence specific to the enzyme TEV protease (EDLYFQ|S). </p>
+
<p>Gas vesicles (GVs) are hollow protein nanostructures synthesized by phototrophic haloarchaea and cyanobacteria to regulate their flotation in aquatic habitats. Bioengineered GVs have been genetically modified for diverse purposes including ultrasonic molecular imaging, gauging cellular turgor pressures, and vaccine delivery — harnessing unique acoustic, mechanical, and surface properties of GVs — but none of their current applications exploits their most fundamental characteristic: buoyancy. </p>
  
−
<p>When the cells have overexpressed enough of the recombinant protein, we will lyse the cells and obtain a protein extract. Since the recombinant protein of interest is fused to a gas vesicle, it will float to the surface of the protein extract and can be skimmed off. After the desired level of purity is obtained (by repeated resuspension and skimming), TEV protease can be added to cleave the recombinant protein of interest from the gas vesicle, dispersing it into the solution while the pure gas vesicles remain at the surface.</p>
+
<p>Our modelling indicates that clusters of GVs float several orders of magnitude better than individual GVs, as buoyancy scales with volume while Stokes’ drag scales with effective radius. Our project iFLOAT aims to improve the flotation of gas vesicles by clustering them using three distinct methods — charge-based flocculation, biotin-streptavidin interaction, and SpyCatcher-SpyTag heterodimerization — and simultaneously develop robust, reproducible flotation assays. Potential future applications of buoyant clusters of bioengineered gas vesicles include bioremediation of oil spills and flotation-based separation and purification of specific targets from mixtures.</p>
  
−
<p>For our project, we will demonstrate a proof of concept using the recombinant protein sfGFP, which we will tag to gvpC. Following this, we will demonstrate the same system in Pichia pastoris to show that eukaryotic protein purification can also be simplified by our method.</p>
 
 
</div>  
 
</div>  
  
−
<div class="clear"></div>
 
−
<div class="column half_size" >
 
−
<h5> Uploading pictures and files </h5>
 
−
<p> You can upload your pictures and files to the iGEM 2017 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. <br />
 
−
When you upload, set the "Destination Filename" to <br><code>T--YourOfficialTeamName--NameOfFile.jpg</code>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)<br><br>
 
−
 
−
<a href="https://2017.igem.org/Special:Upload">
 
−
UPLOAD FILES
 
−
</a>
 
−
</p>
 
−
</div>
 
  
  

Revision as of 13:00, 2 September 2017

Template:Team:IISc-Bangalore

iFLOAT

a multifaceted approach to cluster bioengineered gas vesicles in vitro and enhance their flotation

Gas vesicles (GVs) are hollow protein nanostructures synthesized by phototrophic haloarchaea and cyanobacteria to regulate their flotation in aquatic habitats. Bioengineered GVs have been genetically modified for diverse purposes including ultrasonic molecular imaging, gauging cellular turgor pressures, and vaccine delivery — harnessing unique acoustic, mechanical, and surface properties of GVs — but none of their current applications exploits their most fundamental characteristic: buoyancy.

Our modelling indicates that clusters of GVs float several orders of magnitude better than individual GVs, as buoyancy scales with volume while Stokes’ drag scales with effective radius. Our project iFLOAT aims to improve the flotation of gas vesicles by clustering them using three distinct methods — charge-based flocculation, biotin-streptavidin interaction, and SpyCatcher-SpyTag heterodimerization — and simultaneously develop robust, reproducible flotation assays. Potential future applications of buoyant clusters of bioengineered gas vesicles include bioremediation of oil spills and flotation-based separation and purification of specific targets from mixtures.


Template:Team:IISc-Bangalore/IISc17 footer