Team:UCAS-China/Engineering

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Engineering Success

After brainstorming, we proposed the idea of microbial therapy for H. pylori. We believe that the three most important parts of medical microbiology are drug-related metabolic pathway, targeted gene design and biosafety module. In addition, the hardware and mathematical model should be considered in the complete treatment plan. Therefore, we also designed a mathematical model that can accurately control the drug administration and determine the situation of H. pylori infection in the stomach to assist in the formulation of personalized treatment plan.

We chose the combination of pfnrs and LL-37 as the drug metabolism pathway. Pfnrs was experimentally validated in our project last year and successfully expressed downstream CD47 nanoantibody under hypoxia.

LL-37 is an antibacterial peptide produced by the human immune system. According to literature research, it can be confirmed that it has inhibitory effect on H. pylori, and can be expressed in prokaryotic expression system. On the basis of last year's plasmid, we changed the gene and added the start codon and stop codon. Due to the epidemic situation, we can not finish the experiment before wiki freeze, so we plan to carry out experiments in the future to prove the feasibility of its expression and the antibacterial effect of antimicrobial peptides.

Through previous experiments and literature research, we can prove that this part of the project is successful.

For the design of ammonia sensor in engineering bacteria, we selected the transcription factor NtrC as the NOT gate protein switch, which combined with another downstream NOT gate protein to form an induction switch that can sense the change of ammonia concentration in the environment and turn on the downstream gene transcription. The related design of NtrC is verified by experiments in UCAS-China 2017 project. According to the new literature research and the proof of logic circuit, we can verify the engineering success of this gene circuit.

For the biosafety switch, we creatively connect the toxin-antitoxin system controlled by the cold-inducible on-switch with the "super DOC".Decline of ambient temperature can turn on the translation of DOC toxin protein and kill cells. The existence of degradation tag greatly reduces the possibility of the leaky expression, and improves the treatment efficiency. The toxin-antitoxin system controlled by temperature-sensitive switch was verified by experiments in last year's project, and it has high reliability. As a new design of this year's project, the new original of DOC toxin protein gene with SsrA degradation tag, after the theoretical proof and literature research, can preliminarily verify the success of the project. In the future, we will directly prove the correctness and reliability of such designs through experiments.