Team:Penn/LightActivatedLysis

From 2012.igem.org

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<b>Figure 7</b><br>
<b>Figure 7</b><br>
Figure 7: The production of clyA-his in BL21 in both bacteral lysate and culture medium</div></div>  
Figure 7: The production of clyA-his in BL21 in both bacteral lysate and culture medium</div></div>  
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Revision as of 00:13, 27 October 2012

Penn 2012 iGEM Wiki

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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 37°C until visible colonies were present (~12 hours). Plates were then grown at 25°C under either inducing or non-inducing conditions for 24 hours and imaged. These results are visible in Figure 4.


pDawn-mCherry Dark

pDawn-mCherry Light


pDawn-His-ClyA Dark

pDawn-His-ClyA Light
Figure 4
Spatial Cell Lysis


Figure 5
Figure 5: Colony Spatial Control.

Figure 6
Figure 6: Penn iGEM spatial control
Verification of Expression of Cytolysin A (ClyA)



Figure 7
Figure 7: The production of clyA-his in BL21 in both bacteral lysate and culture medium

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