Team:Penn/LightActivatedLysis
From 2012.igem.org
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.
![](https://static.igem.org/mediawiki/2012/4/42/PDawn-mCherry-Dark-Grey-BG.jpg)
pDawn-mCherry Dark
![](https://static.igem.org/mediawiki/2012/c/c0/PDawn-mCherry-Light-Grey-BG.jpg)
pDawn-mCherry Light
![](https://static.igem.org/mediawiki/2012/d/db/PDawn-ClyA-His-Dark-Grey-BG.jpg)
pDawn-His-ClyA Dark
![](https://static.igem.org/mediawiki/2012/3/35/PDawn-ClyA-His-Light-Grey-BG.jpg)
pDawn-His-ClyA Light
Figure 4
Spatial Cell Lysis
![](https://static.igem.org/mediawiki/2012/7/76/Colony-Spatial-Lysis---Grey-BG.jpg )
Figure 5
Figure 5: Colony Spatial Control.
![](https://static.igem.org/mediawiki/2012/1/10/Penn-iGEM-Lysis---Grey-BG.jpg)
Figure 6
Figure 6: Penn iGEM spatial control
Verification of Expression of Cytolysin A (ClyA)
![](http://partsregistry.org/wiki/images/3/3a/Gel_pic_pdawn.png)
Figure 7
Figure 7: The production of clyA-his in BL21 in both bacteral lysate and culture medium
![](https://static.igem.org/mediawiki/2012/4/4f/HEK293T_Plot.jpg)