Team:Think Edu China/Partnership
Overview
While working on the project, our team DEBIOTICS embraced one of the main value of IGEM: meet with other teams and learn from them to spark more innovative inspirations.
Despite of COVID-limitation, we were still able to work closely with the other two high school teams: LZU-HS-China and XHD-Wuhan-Pro-China. Fortunately, we were able meet and to work in the same lab station in Shanghai XiaoTong University. We are graceful to the platform IGEM had provided, allowing us to contribute and engage with different people who are enthusiastic in the world of synthetic biology. Here is a record of our partnership being done throughout the course of our project.
Part1: Project design with XHD-Wuhan-Pro-China
After determine the direction of our project, degradation of antibiotics, we were impeded to choose an appropriate cell carrier. XHD_Wuhan_Pro_China inspired us with their project which is degrade alcohol consumption, specifically coincidentally their project also construct an intestinal probiotics. After discuss with our project instructor, Professor Liu, they suggest us and provided us the strain of Nissle 1917 (EcN). After the discussion, we have done some research on Nissle 1917, and found out that this probiotic has been widely used in synthetic engineering. The nature of EcN makes it easy to carry other foreign recombinant factors, also proven to be harmless.
This selection of appropriate strain advanced our project and we were ready to move on the the next stage of constructing an engineered strain for the degradation of antibiotics.
Figure 1. Experimental Data from XHD-Wuhan-Pro-China
Figure 2. Group picture of XHD-Wuhan-Pro-China
Their wiki link : https://2021.igem.org/Team:XHD-Wuhan-Pro-China
LZU-HS-China
During the process of discussion and communication, the other high school team, LZU_HS_China joined the laboratory.
After two weeks of investigate, their group discovered that because EcN contains secret plasmids, it can interfere with the expression of foreign genes at specific locations to some extent and disturbed the efficacy of the gene and protein. Thus, their project by adopt a new plasmid-PMB1A3. Moreover, the strong promoter before the expressive sequence and INP, an appropraite and stable protein transporter binding on cell membrane, were added in front of the target gene to promote the efficiency and stability.
LZU_HS_China's investigation helped us to avoid potential deficiencies in our experiments, their research result of the INP protein was further used in our experiment as its nature allowed the laccase enzyme to be fixed on cell surface to increase the efficiency of antibiotics degradation.
Influence on our design:
LZU-HS-China team helps us design of the plasmid for our project as we shared the result of their investigation on the INP-N protein. We inserted the gene fragment of the INP-N, Lacc6, and GEP to the plasmid to construct the engineering bacterial strain.
Their WIKI: https://2021.igem.org/Teamm:LZU-HS-China
Think Edu China (Cell Surface display technology)
When our team are reconsidering the value of the project, we are thinking what are some ways that can be more effective that distinguish ourselves from the conventional methods such as membrane bioreactor and selected wastewater treatment plants.
Through research and discussion with our project instructor, instead of expressing our enzymes and proteins within the cell membrane, we figure out another feasible way to express our foreign functional protein. We applyed the microbial cell-surface display technology to make our project more economically and effectively advantageous. We discovered that this system can also enhance the stability and efficiency of reactive enzymes as well as prevent the degradation of foreign factors by intracellular enzymes produced by the host.
The cell surface display technology is a suitable solution for the SAs degradation; this system prevent the effect of before the start of degradation, the bacterial cells could be killed by the antibiotics. This system allows the laccase enzyme to bind to the cell surface and do its job.
Figure 6. Cell surface display system adopted by LZU-HS-China team in designing their project.
We shared our discovery with the other two high school teams, LZU-HS-China adopted this idea as well since their project is similar. The construction of a novel whole cell biocatalyst that exhibits the selenite reductive SerV01 on the cell surface of EcN as a new engineered bacterium (EcN-IS).
Wiki link: https://2021.igem.org/Team:Think_Edu_China
Final Stage
The three teams worked together at SJTU for two weeks to do the wet lab experiment. After the designing the project, we were able to express the laccase in vitro, and successfully to see the degradation of Sulfonamide antibiotics.
