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| class="logo"> | | class="logo"> |
| </div> | | </div> |
− | <p>EpiFlex aims to make the bacterium a chassis for synthetic biology. | + | <p>EpiFlex aims to make the <i>Staphylococcus epidermidis</i> bacterium a chassis for synthetic biology. |
| </p> | | </p> |
| | | |
| <p><b class="heading">MoClo Toolkit</b></p> | | <p><b class="heading">MoClo Toolkit</b></p> |
− | <p>The MoClo is a modular cloning method based on Golden Gate assembly. EpiFlex is a Moclo | + | <p>MoClo is a modular cloning method based on Golden Gate assembly. EpiFlex is a Moclo |
| toolkit developed with parts that function specifically in S.epidermidis. | | toolkit developed with parts that function specifically in S.epidermidis. |
| </p> | | </p> |
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| <!--Write the text explaining this section --> | | <!--Write the text explaining this section --> |
| <div class="text"> | | <div class="text"> |
− | <p>EpiGlow is the proof of concept of our EpiFlex toolkit. We choose to express mCherry | + | <p>EpiGlow is the proof of concept of our EpiFlex toolkit. We chose to express mCherry |
| in <i>S. epidermidis</i> to demonstrate that our parts are functional. | | in <i>S. epidermidis</i> to demonstrate that our parts are functional. |
| </p> | | </p> |
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| | | |
| <ul> | | <ul> |
− | <li>We explored optimised electroporation protocols<sup>1</sup> by testing different voltages and a | + | <li>We explored and optimized electroporation protocols<sup>1</sup> by testing different voltages and a |
| newer heat shock/ electroporation combination<sup>2</sup>, to see which protocol would yield | | newer heat shock/ electroporation combination<sup>2</sup>, to see which protocol would yield |
| more transformants. | | more transformants. |
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| <p><b class="heading">Results</b></p> | | <p><b class="heading">Results</b></p> |
| <ul> | | <ul> |
− | <li>Using our EpiFlex system we were able to successfully build a construct that | + | <li>Using our EpiFlex system, we were able to successfully build a construct that |
| expressed mCherry in <i>S. epidermidis</i> | | expressed mCherry in <i>S. epidermidis</i> |
| </li> | | </li> |
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| | | |
| <ul> | | <ul> |
− | <li>In our optimisation of the electroporation of <i>S. epidermidis</i>, we found that a | + | <li>In our optimization of the electroporation of <i>S. epidermidis</i>, we found that a |
| voltage of 2.5kV yielded the greatest transformation efficiency. We achieve around 3 | | voltage of 2.5kV yielded the greatest transformation efficiency. We achieve around 3 |
| - 7 transformants per plate using this protocol. | | - 7 transformants per plate using this protocol. |
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| temperatures as low as 33°C (hands, feet, nose), up to 40°C (severe conditions). | | temperatures as low as 33°C (hands, feet, nose), up to 40°C (severe conditions). |
| </p> | | </p> |
− | <p>For each temperature, the growth was measured in TSB for 15 h via optical density. | + | <p>At each temperature, the growth was measured in TSB for 15 hours via optical density measurements. |
| </p> | | </p> |
| <p><b>2. Acidity</b></p> | | <p><b>2. Acidity</b></p> |
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| genital area) depending on the region of the body it is covering. | | genital area) depending on the region of the body it is covering. |
| </p> | | </p> |
− | <p>We tested pH range from pH3 to pH10. The measurement has been done in the same way as for | + | <p>We tested in pH range from pH3 to pH10. The measurement has been done in the same way as for |
| the Temperature tests. | | the Temperature tests. |
| </p> | | </p> |
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| <p>With respect to the skin, there can be variations in salinity caused mainly by sweat and | | <p>With respect to the skin, there can be variations in salinity caused mainly by sweat and |
| sebaceous glands. The evaporation of water from the release of heat enables the salts to | | sebaceous glands. The evaporation of water from the release of heat enables the salts to |
− | remain present on the skin. | + | remain on the skin. |
| </p> | | </p> |
− | <p>We made vary the salinity of the media from 0.5% to 5.5% of NaCl. Again, the measurements | + | <p>We varied the salinity of the media from 0.5% to 5.5% of NaCl. Again, the measurements |
− | have been done over 15h of growth in TSB. | + | have been done over 15 hours of growth in TSB. |
| </p> | | </p> |
| | | |
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| </b></p> | | </b></p> |
| <ul> | | <ul> |
− | <li>Improve transformation efficiency form electroporation | + | <li>Improve transformation efficiency from electroporation |
| </li> | | </li> |
| <li>Build more constructs using the EpiFlex toolkit to characterize all the parts | | <li>Build more constructs using the EpiFlex toolkit to characterize all the parts |
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| </b></p> | | </b></p> |
| <ul> | | <ul> |
− | <li>Using EpiFlex to build a genetic circuit aiming to control population dynamic in the | + | <li>Using EpiFlex to build a genetic circuit aiming to control population dynamics in the |
| skin microbiome. | | skin microbiome. |
| </li> | | </li> |
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| <p><b class="heading">EpiGrow</b></p> | | <p><b class="heading">EpiGrow</b></p> |
| <ul> | | <ul> |
− | <li>We would like to investigate the difference of growth kinetic between <i>S. epidermidis</i> | + | <li>We would like to investigate the difference of growth kinetics between <i>S. epidermidis</i> |
| grown in 2D and in 3D media. | | grown in 2D and in 3D media. |
| </li> | | </li> |
− | <li>To do so, we decided to build an artificial skin model and develop a method to | + | <li>We decided to build an artificial skin model of the human palm to develop a method to |
| measure growth on 2D media while having comparable data with growth in 3D media. | | measure growth on 2D media while having comparable data with growth in 3D media. |
− |
| |
| </li> | | </li> |
| </ul> | | </ul> |