Difference between revisions of "Team:IISc-Bangalore"

 
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{{Team:IISc-Bangalore}}
 
 
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<h1> iFLOAT: Isolation of Floater-Linked Overexpressed-protein Ascender Technology </h1>
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<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>
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<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>
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          <div style="position: absolute; right: 5%; width: 35%; font-size: 2.3em; line-height: 1.4; font-family: 'Poppins'; top: 50%; transform: translateY(-50%);">Gas vesicles (GVs) are hollow protein nanostructures synthesized by phototrophic haloarchaea and cyanobacteria to regulate their flotation in aquatic habitats. </div>
  
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<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>
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<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>
 
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<h5> Uploading pictures and files </h5>
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<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 />
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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>
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UPLOAD FILES
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          <div style="position: absolute; left: 3%; width: 33%; font-size: 2.3em; line-height: 1.4; font-family: 'Poppins', sans-serif; top: 55%; transform: translateY(-50%);">Bioengineered GVs have been genetically modified for diverse purposes; ultrasonic molecular imaging, gauging cellular turgor pressures, and vaccine delivery - but none of their current applications exploit their most fundamental characteristic: buoyancy.</div>
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          <div class="gifAnimated" style="background-image: url('https://static.igem.org/mediawiki/2017/4/45/T--IISc-Bangalore--home-3.gif'); background-position: left;"></div>
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          <div style="position: absolute; right: 5%; width: 20%; font-size: 2.3em; line-height: 1.4; font-family: 'Poppins'; top: 55%; transform: translateY(-50%);">Our modelling shows that clusters of GVs float enormously better than individual GVs.</div>
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          <div class="gifAnimated" style="background-image: url('https://static.igem.org/mediawiki/2017/7/71/T--IISc-Bangalore--home-4.gif'); background-position: right;"></div>
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          <div style="position: absolute; left: 3%; width: 30%; font-size: 2.3em; line-height: 1.4; font-family: 'Poppins', sans-serif; top: 55%; transform: translateY(-50%);">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</div>
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          <div style="position: absolute; right: 5%; width: 25%; font-size: 2.3em; line-height: 1.4; font-family: 'Poppins'; top: 55%; transform: translateY(-50%);"> Future applications of these clusters can include bioremediation of oil spills, flotation-based separation, and purification of specific targets from mixtures!</div>
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              <div style="font-size: 2.5em; font-family: 'Raleway-ExtraBold', sans-serif; margin-top: 20px; text-align: center; color: black;" class='textclass'>ABOUT</div></a>
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              <a href="https://2017.igem.org/Team:IISc-Bangalore/Description"><img src='https://static.igem.org/mediawiki/2017/3/3b/T--IISc-Bangalore--icons-project.svg' width=100% />
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              <div style="font-size: 2.5em; font-family: 'Raleway-ExtraBold', sans-serif; margin-top: 20px; text-align: center; color: black;" class='textclass'>PROJECT</div></a>
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              <a href="https://2017.igem.org/Team:IISc-Bangalore/Assembly"><img src='https://static.igem.org/mediawiki/2017/6/6f/T--IISc-Bangalore--icons-lab.svg' width=100% />
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Latest revision as of 03:41, 2 November 2017

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; ultrasonic molecular imaging, gauging cellular turgor pressures, and vaccine delivery - but none of their current applications exploit their most fundamental characteristic: buoyancy.
Our modelling shows that clusters of GVs float enormously better than individual GVs.
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
Future applications of these clusters can include bioremediation of oil spills, flotation-based separation, and purification of specific targets from mixtures!