"
Team:Penn/LightActivatedLysis
From 2012.igem.org
(Created page with "{{:Team:Penn/Template/Site}} <html> <div style="background-color:white;padding-left:20px; padding-right:20px;"> <h1><b>Light-Dependent Lysis of Mammalian Cells by Bacteria</b...") |
|||
| Line 4: | Line 4: | ||
| - | <div style=" | + | <div style="padding-left:20px; padding-right:20px;"> |
<h1><b>Light-Dependent Lysis of Mammalian Cells by Bacteria</b></h1> | <h1><b>Light-Dependent Lysis of Mammalian Cells by Bacteria</b></h1> | ||
Revision as of 04:45, 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
