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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.
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
