Team:DLUT China/Demonstrate

From lab to the real world

D emonstrate

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1
We intend to use the lethal system 1 we designed and decided to introduce them into the Nissle E. coli strain, which is shown below.

2
Due to the limitations of the experimental conditions (we lack experimental materials and equipment that can simulate the intestinal environment), we only performed a demonstration of the lethal system.
We also determined the feasibility of inducing a lethal system in vitro. We collaborated with Northeastern University to verify the cold lethal system induced by pCold (using toxic proteins mazF) and the results are shown below.

Boom !

3

Our data indicate that due to the high toxicity of mazF, even low expression of toxic proteins is sufficient for cell death. In contrast, the cold shock expression Lysis showed a better kill switch property. In vivo at 37℃, low expression of lysis is not enough to cause functional probiotics to die, while short-term (4h) cold shock induction in vitro can fully inhibit cell growth. In short, cold shock expression of Lysis as a kill switch is a better choice.

4

However, there are many limitations in inducing lethality in vitro, such as the induction of 16 degrees Celsius and the need to add inducers, so this lethal method is not a perfect method of lethality.
We plan to add our recombinant strains to fermented beverages or to make biobacteria for patients to take. However, due to the influence of time and other factors, the work of this part has not been completed, we plan to follow up on this part of the work.