Contribution | Thinker-SC

1、inclusivity：

We believe that from the aspect of inclusivity we can help future IGEM teams in one aspect. The main thing in that area is innovation. Why do we say this? The main reason is that in terms of the theme of our "inclusive" section, the main content and people we target are people with Tourette syndrome, and our purpose is to help them through our planned actions and let society pay more attention to them. , give them more respect and attention. The reason is that Tourette's syndrome is often not mentioned in various current diseases, and it is also unpopular. Society will naturally ignore it when comparing other diseases. In addition, in the previous inclusive section of IGEM, almost no group had a project on Tourette syndrome. Almost everyone made reference and modifications based on previous excellent projects, so the theme did not change much. Therefore, we want to innovate. We will not draw on the themes and content based on the excellent projects in the "Inclusion" section. We think we will help Tourette in society through our own plans and our own actions. disease patients. We also hope that in the future, the IGEM team can also create more different and unpopular themes, including diseases and social stereotypes, just like Tourette's syndrome, so that more people can understand it. Recognizing it, understanding it, paying attention to it, and helping it are also our original intention and purpose for the entire "inclusive" section.

Therefore, in order to understand the public’s understanding of the Torrey Syndrome community, our team conducted an online questionnaire survey（seen in figure 1

figure 1 the survey report

The pie chart clearly show that only a few people in China understand or know what is Torre's Syndrome.This also makes our team more determined to help these patients with Torre's Syndrome so that they can be treated equally like ordinary people. We first found patients with Torre's syndrome through social media channels, introduced what igem is and our team, and arranged a time for online interviews, from which we learned a lot more about Torre's syndrome. . Picture 2 shows our team conducting an online meeting with patients

figure 2 part of interview

After communicating online with many patients, we compiled the content of our interviews and decided to create a platform to help the public understand Torre's syndrome.

figure 3 A social platform we created ourselves

We have published five articles since our creation and have received a lot of attention.

In the future, we plan to cooperate with relevant Torre's syndrome organizations and create our own science classes to help them learn.We also hope that the future igem team can, like us, focus more on these unpopular things that are easily ignored by the public. Let more people and problems get help and solutions

2,technological innovation：

biological crust

In order to make our products more innovative and create a good growth environment for seeds to promote seed germination, we use biological crusting to achieve the above purposes. To achieve biological crusting, the production of extracellular polysaccharides (EPS) is necessary. Extracellular polysaccharides play a key role in bacterial soil crusting. They can provide microbial adhesion to soil particles and help microorganisms grow and colonize better on the soil surface. However, the level of EPS will be low in areas with severe soil erosion. The low level of EPS can be improved by changing the levels of central metabolic enzymes that affect the production of precursor nucleotide sugars. We identified and cloned the gaIu gene encoding UPD glucose pyrophosphorylase (GaIU) in Streptococcus thermophilus LY03 to enhance EPS production

The gaIU gene is a gene encoding UDP glucose pyrophosphorylase, which exists in a variety of organisms, including Streptococcus thermophilus LY03.

We plan to overexpress the galU gene using the T7 promoter and the B0015 terminator in the pET23b vector. The constructed plasmid will then be transformed into E. coli Rosetta (host cells). A schematic diagram illustrating this process can be seen in figure 1

figure 1 the design of gaIU

figure 2 Gel electrophoresis of the galU.

Characterization

The successfully engineered strain will be expanded in culture.At predetermined time points (4, 8, 12, 16 hours), bacterial growth was monitored by measuring the OD600. Concurrently, cells were centrifuged, and the supernatant was used to quantify EPS using the anthrone-sulfuric acid method. In brief, a sample of the supernatant was combined with anthrone reagent, followed by the addition of concentrated sulfuric acid. The mixture was subsequently heated, and the developed color was measured spectrophotometrically at 620 nm. A standard curve, established using known concentrations of a standard polysaccharide, facilitated the determination of the EPS concentration in the samples. The EPS concentration was then normalized to the OD600 to obtain EPS production per unit of bacterial growth. To establish a standard curve, 0-200 mM glucose was used as the standard substance and quantified using the anthrone colorimetric method, with absorbance measured at 620 nm, as shown in Figure 3A. Compared to the control group, the engineered bacteria significantly increased EPS levels, which also increased with time, as shown in Figure 3B. The results indicate that the engineered strain can significantly enhance EPS production.

Figure 3 The expression effect of galU.

future contribution

Our biological crust not only provides a good living environment for microorganisms in the soil, but more importantly, it lays a foundation for the future igem team, who can think of more efficient and effective methods based on our biological crust. To solve soil erosion problems or soil problems in order to protect vegetation and trees in nature.For example, our team is currently using the gaIU gene to increase EPS production. Our ideas can serve as a reference for future teams. Maybe they will think of other more effective and low-cost methods to increase EPS production. This can greatly save their time and cost, allowing them to spend more time on innovation.