Line 35: | Line 35: | ||
We measured the growth rate of cells producing different chromoproteins by using a plate reader to measure the optical density of isolated populations over 10 hours In order to determine if different types of chromoprotein expression were influencing the growth rate differently.<br><br> | We measured the growth rate of cells producing different chromoproteins by using a plate reader to measure the optical density of isolated populations over 10 hours In order to determine if different types of chromoprotein expression were influencing the growth rate differently.<br><br> | ||
− | We want to show that growth control can be used to produce a stable co-culture that can maintain its ratio overtime. Therefore we combined the arabinose-inducible | + | We want to show that growth control can be used to produce a stable co-culture that can maintain its ratio overtime. Therefore we combined the arabinose-inducible Gp2 construct with a construct for the chromoprotein eforRed. We then induced the cells with arabinose to observe the effect of Gp2 on the population stability of the colored cells.<br><br> |
Line 54: | Line 54: | ||
</center> | </center> | ||
− | <p>The cells transformed with the efoRed+ | + | <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> | ||
<center> | <center> | ||
<img src="https://static.igem.org/mediawiki/2016/a/ae/T--Imperial_College--Proof3.png" height="470"/><br> | <img src="https://static.igem.org/mediawiki/2016/a/ae/T--Imperial_College--Proof3.png" height="470"/><br> | ||
− | <p><b>Figure 3:</b> Plot of Growth rate versus time for the eforRed with growth control | + | <p><b>Figure 3:</b> Plot of Growth rate versus time for the eforRed with growth control Gp2 construct <i> Escherichia coli</i> Top 10 cells. </p> |
</center> | </center> | ||
Line 65: | Line 65: | ||
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 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+ | + | 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> |
Revision as of 00:56, 20 October 2016