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<specialh3>Colour proof</specialh3><br><br> | <specialh3>Colour proof</specialh3><br><br> | ||
<p> | <p> | ||
− | In order to demonstrate one of the many possible applications of our ratio control circuit, we decided to use chromoproteins as visual proof of concept. We were able to create a multitude of colours by mixing different ratios of <i>E. coli</i> cultures expressing the chromoproteins. We hope that this provided a glimpse of the power of co-culture in the production of composite biomaterials. <br> | + | In order to demonstrate one of the many possible applications of our ratio control circuit, we decided to use chromoproteins as visual proof of concept. We were able to create a multitude of colours by mixing different ratios of <i>E. coli</i> cultures expressing the chromoproteins. We hope that this provided a glimpse of the power of co-culture in the production of composite biomaterials. <br><br> |
<specialh3>Key Achievements</specialh3><br><br> | <specialh3>Key Achievements</specialh3><br><br> | ||
Built constructs for expressing 7 different chromoproteins <br> | Built constructs for expressing 7 different chromoproteins <br> | ||
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<br><br><specialh3>Results</specialh3><br><br> | <br><br><specialh3>Results</specialh3><br><br> | ||
In our cell mixing experiment, we were able to produce over 70 different colors from the 7 base chromoproteins, indicating that a wide range of biological colours can potentially be produced by chromoprotein mixing. <br><br> | In our cell mixing experiment, we were able to produce over 70 different colors from the 7 base chromoproteins, indicating that a wide range of biological colours can potentially be produced by chromoprotein mixing. <br><br> | ||
− | + | </p> | |
<center> | <center> | ||
<img src="https://static.igem.org/mediawiki/2016/a/a6/T--Imperial_College--Proof1.png" height="500"/><br> | <img src="https://static.igem.org/mediawiki/2016/a/a6/T--Imperial_College--Proof1.png" height="500"/><br> | ||
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</center> | </center> | ||
− | The cells transformed with the efoRed+GP2 construct showed a decrease in growth rate when induced with arabinose, suggesting that our circuit can be a suitable system for controlling the growth of colored cells.</p> | + | <p>The cells transformed with the efoRed+GP2 construct showed a decrease in growth rate when induced with arabinose, suggesting that our circuit can be a suitable system for controlling the growth of colored cells.</p> |
<br><br> | <br><br> | ||
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</center> | </center> | ||
− | <br><br><specialh3>What's Next?</specialh3><br><br> | + | <p><br><br><specialh3>What's Next?</specialh3><br><br> |
− | We aim to demonstrate different chromoprotein expressing cells growing together in co-culture. To do this, we plan on inoculating the two populations at a 1:1 ratio and recording the peak absorption over time. This would allow us to observe if the population composition is changing, as it would be reflected in a change in color and therefore in absorption properties. We could then determine if the slower growing population, as determined from our earlier growth rate experiments, is outcompeted by the other population as expected.<br> | + | We aim to demonstrate different chromoprotein expressing cells growing together in co-culture. To do this, we plan on inoculating the two populations at a 1:1 ratio and recording the peak absorption over time. This would allow us to observe if the population composition is changing, as it would be reflected in a change in color and therefore in absorption properties. We could then determine if the slower growing population, as determined from our earlier growth rate experiments, is outcompeted by the other population as expected.<br><br> |
− | We also hope to grow the cells transformed with the efoRed+GP2 under the control of the arabinose promoter alongside cells expressing the chromoprotein amajLime. We could then monitor the absorption of the sample after induction with different amounts of arabinose to see if varying levels of induction changed the colour of the co-culture. | + | We also hope to grow the cells transformed with the efoRed+GP2 under the control of the arabinose promoter alongside cells expressing the chromoprotein amajLime. We could then monitor the absorption of the sample after induction with different amounts of arabinose to see if varying levels of induction changed the colour of the co-culture. |
Once these preliminary experiments are successfully completed, we hope to integrate different chromoprotein genes into the genomes of two populations of <i> E. coli</i> and transform our circuit plasmids into the cells. We would then induce the co-culture with different ratios of AHLs to set various population ratios, which would be visible as different colours. <br><br> | Once these preliminary experiments are successfully completed, we hope to integrate different chromoprotein genes into the genomes of two populations of <i> E. coli</i> and transform our circuit plasmids into the cells. We would then induce the co-culture with different ratios of AHLs to set various population ratios, which would be visible as different colours. <br><br> | ||
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<specialh3>Experience</specialh3><br><br> | <specialh3>Experience</specialh3><br><br> | ||
− | Chromoproteins allow easy identification of successfully transformed cells. However different chromoproteins have different maturation time and provide different colour intensity. This further complicate the mixing process as more intense colours are better at being mixed that the less intense ones. <br> | + | Chromoproteins allow easy identification of successfully transformed cells. However different chromoproteins have different maturation time and provide different colour intensity. This further complicate the mixing process as more intense colours are better at being mixed that the less intense ones. <br></p> |
Revision as of 21:13, 19 October 2016