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Team:Penn/LightActivatedLysis
From 2012.igem.org
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| + | <h1 align="center"><b>Light-Dependent Lysis of Mammalian Cells by Bacteria</b></h1><br /> | ||
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We then wanted to prove that our pDawn-ClyA construct was able to lyse mammalian cells in a light-dependent manner. To assess this, we plated BL21 bacteria transformed with pDawn-ClyA or pDawn-mCherry on Columbia Agar plates supplemented with 5% Sheep Blood (BD). These plates are used to qualitatively detect hemolytic activity in bacteria by visually confirming lysis through a color change in the media as the blood cells are lysed. After plating the bacteria, cultures were grown in non-inducing conditions at 37C until visible colonies were present (~12 hours). Plates were then grown at 25C under either inducing or non-inducing conditions for 24 hours and imaged. These results are visible in Figure 2.</p> | We then wanted to prove that our pDawn-ClyA construct was able to lyse mammalian cells in a light-dependent manner. To assess this, we plated BL21 bacteria transformed with pDawn-ClyA or pDawn-mCherry on Columbia Agar plates supplemented with 5% Sheep Blood (BD). These plates are used to qualitatively detect hemolytic activity in bacteria by visually confirming lysis through a color change in the media as the blood cells are lysed. After plating the bacteria, cultures were grown in non-inducing conditions at 37C until visible colonies were present (~12 hours). Plates were then grown at 25C under either inducing or non-inducing conditions for 24 hours and imaged. These results are visible in Figure 2.</p> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/1/14/Figure3_pDawn.PNG" width="500" | + | <div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/1/14/Figure3_pDawn.PNG" width="500"><br> |
| - | < | + | <b>Figure 4</b></div></div> |
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| + | <h1 align="center"><b>pDawn and Nissle 1917</b></h1><br /> | ||
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<p style="color:black;text-indent:30px;"> | <p style="color:black;text-indent:30px;"> | ||
In order to further develop our system for future in vivo therapeutic applications, we transformed Nissle 1917 with pDawn-mCherry to see if we could implement our system into a non-pathogenic strain of E. coli. We repeated our initial experiments and achieved light-dependent gene expression in Nissle 1917 for the first time ever. We are now hoping to clone in our pDawn-ClyA construct to show that Nissle 1917 is capable of light-dependent lysis of mammalian cells. Stay tuned! | In order to further develop our system for future in vivo therapeutic applications, we transformed Nissle 1917 with pDawn-mCherry to see if we could implement our system into a non-pathogenic strain of E. coli. We repeated our initial experiments and achieved light-dependent gene expression in Nissle 1917 for the first time ever. We are now hoping to clone in our pDawn-ClyA construct to show that Nissle 1917 is capable of light-dependent lysis of mammalian cells. Stay tuned! | ||
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| - | <img src="https://static.igem.org/mediawiki/2012/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="250" height="350" | + | <div class="fig"><div align="center"><img src="https://static.igem.org/mediawiki/2012/7/72/Nissle-1917-pDawn-mCherry-10-1-2012.jpg" width="250" height="350"><br> |
| - | < | + | <b>Figure 5</b></div></div> |
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Revision as of 14:39, 22 October 2012
Light-Dependent Lysis of Mammalian Cells by Bacteria
We then wanted to prove that our pDawn-ClyA construct was able to lyse mammalian cells in a light-dependent manner. To assess this, we plated BL21 bacteria transformed with pDawn-ClyA or pDawn-mCherry on Columbia Agar plates supplemented with 5% Sheep Blood (BD). These plates are used to qualitatively detect hemolytic activity in bacteria by visually confirming lysis through a color change in the media as the blood cells are lysed. After plating the bacteria, cultures were grown in non-inducing conditions at 37C until visible colonies were present (~12 hours). Plates were then grown at 25C under either inducing or non-inducing conditions for 24 hours and imaged. These results are visible in Figure 2.
Figure 4
pDawn and Nissle 1917
In order to further develop our system for future in vivo therapeutic applications, we transformed Nissle 1917 with pDawn-mCherry to see if we could implement our system into a non-pathogenic strain of E. coli. We repeated our initial experiments and achieved light-dependent gene expression in Nissle 1917 for the first time ever. We are now hoping to clone in our pDawn-ClyA construct to show that Nissle 1917 is capable of light-dependent lysis of mammalian cells. Stay tuned!
Figure 5
