Initial stage of the project
At the beginning of the project, the teams of both parties initially determined the direction of the project, they contacted each other and conducted preliminary exchanges and discussions, introduced their respective inspirations and preliminary plans, and respectively put forward their opinions on the overall idea. Since the two teams are in Beijing, and the projects of the two teams are very closely related and sought together, BIT-China and BUCT-Chinahave developed the idea of long-term cooperation between the two teams.
At the beginning of the project, the two teams discussed all aspects of the project:
Project mid-term communication
After a period of experimentation, the two parties made slight changes to their respective project plans based on the degree of the experiment and combined with the newly generated inspiration, and initially established the type of track to be selected.
Because of the impact of the epidemic, everyone held an online meeting.
In the first stage of the meeting, the two teams each introduced their overall project structure. Our theme is "Creative Tasting Officer", and the plan is to construct a biosensor using Saccharomyces cerevisiae as the base cell to detect the ratio of sweetness and some bitter substances in food.The students from Beijing University of Chemical Technology chose the synthetic artificial meat project based on the damage caused by the current animal husbandry to the environment. In addition, detailed discussions were carried out on the current project content.
In this conference, the BIT-China team and the BUCT-China team cooperated to conduct popular science lectures in the form of live broadcast.The students of the BUCT-China team gave a science lecture on artificial meat, explaining to the audience the development process of artificial meat, the synthesis process of cell culture meat, the knowledge of culturing meat and traditional livestock meat, and cultivating meat and plant-based artificial meat. The students of the BIT-China team let the audience systematically understand synthetic biology and explained the basic concepts of synthetic biology in easy-to-understand language, making the emerging discipline of synthetic biology more deeply rooted in the hearts of the people.
In addition, we also discussed how to carry out specific cooperation in social practice, popular science education, art, modeling and experiment.
At the same time, we discussed the content of the project and watched the project publicity video of both parties. Through consultation, we worked out the cooperation plan and implementation rules in five stages of social practice, popular science education, art, modeling and experiment. At the same time, a new idea of constructing food safety evaluation system by using modeling method is put forward.
The following is the exchange experience of the two sides in the mid-term:
The suggestions we provided to BUCT-China:
BUCT-China's artificial meat project involves cell culture, so we have some questions about cell culture. In response, the students of BUCT-China team said that the cells were extracted from chickens about 30 days old and then cultured for 3 generations for differentiation culture. It was estimated that the success rate of the experiment was high.
The suggestions we received from BUCT-China:
BUCT-China proposed that sweet receptors and umami receptors were heterodimers, and the assembly position was intracellular assembly and located on the membrane or directly assembled on the membrane. Bitchina responded that the current experimental scheme was intracellular assembly and located on the membrane, because it was difficult to assemble on the membrane and the experimental success rate was low.
Modeling and prediction
Students in the modeling group preliminatively worked out a specific plan to build the food safety evaluation system. Tianyi Bai from BIT-China shared some information obtained through literature review. The construction of food safety system mostly adopts analytic hierarchy process (AHP) or fuzzy comprehensive evaluation method, and the main indicators are the description of hazard characteristics, intake evaluation, risk characteristics and so on. BUCT-China student Yang Yang proposed, using the analytic hierarchy or food safety evaluation system model is set up priority is we want to get enough true data, therefore, two groups of students for specific planning on modeling work done in the late, two teams need to find their own product evaluation index, after we discuss, To find common indicators and carry out the next step.
In addition, it is worth mentioning that in this conference, students from BUCT-China team shared some of their gains on modeling in the CCiC conference. He proposed that the ideal modeling should be derived from real experiments, and the model should be modified and fitted repeatedly through the experimental conditions and obtained data. Moreover, the model we finally established should be helpful to the experimental model. This point of view facilitated the modeling work of the two teams.
In the cooperation between the students of the social practice groups of the two teams at this stage, the main purpose is to help each other forward the questionnaires and related pushes of the respective teams, in order to obtain richer statistical results and feedback.
Later stage of the project
After consultation with the students of the BIT-China team and the BUCT-China team, everyone thinks that avoiding the large group discussion and adopting the form of group communication is more conducive to the exchange of opinions and can better improve each section. In response to this decision, post-communication is basically conducted online meetings in a group mode.
Because the two teams have already conducted preliminary discussions on some aspects of Human practice in previous meetings. Therefore, when we design social practices, we mainly communicate from the aspects of product research, experimentation, and putting into use. In the communication with COFCO, the biosensor part of our experiment has been recognized by the other party, and it has been evaluated for its broad application prospects. This will be a powerful grasp for the detection part in the later stage of the experiment. The two parties intend to conduct a joint test.
In the later period, most of the experiments on both sides have basically been carried out, and some relatively ideal experimental results have been obtained. But it cannot be denied that everyone still encountered some difficult problems in the experiment. The two sides exchanged views on the difficulties in their experiments and both put forward their own suggestions.
We also conducted exchanges with students in the modeling group, art group, and website group. The students from the two teams expressed their opinions and talked with each other, and everyone benefited.
After getting a more detailed understanding of the experimental schemes of both parties, everyone put forward the idea of sharing the experimental tasks with each other.
The suggestions we provide to BUCT-China:
The most important part of the project for the students at BUCT-China is the polyester synthesis part. The ability to synthesize polyester using the cells themselves is the key to synthesizing the structure of artificial meat fibers. Also, the use of cells to produce polyester materials with a longer carbon skeleton is a new breakthrough.
In this part of the work, in addition to the construction of the intracellular polyester synthesis pathway is the focus of this part of the work, how to extract the product and how to detect it is also a major headache in this part of the work.
In the early stage of the experiment, due to the low yield of the product and the poor solubility of the product, the BUCT-China team used common organic solvents such as hexane, cyclohexane and ethyl acetate for the extraction of the product, followed by gas chromatography for the detection of the product. However, after several attempts, it was found that the solubility of the products in these conventional solvents was not high, so the students of BUCT-China did not get more satisfactory data results in the gas phase in the initial stage.
To solve this problem, we had a discussion with the BUCT-China team, and our students provided the products they speculated might exist,such as: nonanoic acid hydroxydodecanoate, dimeric hydroxydodecanoic acid, etc. After the exchange and discussion, we concluded that extraction of dimeric hydroxydodecanoic acid with conventional organic solvents was not efficient, and we recommended to them the use of trichloromethane for extraction.
In the subsequent experiments, the students of BUCT-China extracted the cell-breaking solution by adding trichloromethane in the ratio of 1:5, and the target products such as nonanoic acid hydroxydodecanoate and dimeric hydroxydodecanoic acid were detected from the gas phase detection.
The suggestions we received from BUCT-China:
We plan to obtain plasmids for the expression of fresh, sweet and bitter human receptors to be used in combination with the detection plasmids we have successfully constructed to measure the taste of food by fluorescence intensity.
In the previous experiments, we took the plasmid PCR to get the linear fragment, and after that, we designed primers and performed PCR to get the fragment of the target gene, and the fragment of the fluorescent gene. The two fragments were ligated by OE technique, and then Gibson ligation of the linear fragment and the OE fragment was performed, which was introduced into E. coli for verification, and if colonies were grown, colony PCR was performed, and the colonies were sequenced after correct PCR.
In this way we successfully constructed the detection plasmid.
However, we encountered difficulties in the construction of the other three plasmids. When we performed linear PCR on the plasmids, the recovery concentration was too low due to the long fragment length, or the PCR did not show bands. In the communication with BUCT-China, we raised this problem, and the BUCT-China students gave us the suggestion that for the low recovery concentration, we can consider running the validation gel first, and if the band is single, then we can perform the column recovery, which will be much higher than the gel recovery concentration. If no band appeared, then consider replacing the primers.
At the same time, they taught us how to use Oligo for primer design, so that the specificity of the designed primers will be better and clear bands can be obtained after PCR.
Since then we have tried the BUCT-China approach and the experiment has a good progress. However, because the linear fragment of the plasmid was too long and not a single band, it could only take the form of gel recovery. We chose fastfu and phanta enzyme systems for PCR in the lab, and tried to change the PCR temperature, but the results were still not promising. We were able to achieve a concentration of 213ng/ul by BUCT-China's PCR.
Using the PCR products they helped us to obtain we successfully passed Gibson and obtained the sweet taste receptor fluorescent plasmid.
When constructing the bitter plasmid, a new problem emerged. We constructed the bitter plasmid in the same way as described above, but the OE always failed to get the correct product, even though the fragment length was correct after the OE run, but there was no way to get the target plasmid when performing Gibson. Using Oligo to design primers or changing the PCR temperature also failed to solve this problem.
We talked with BUCT-China's experimental group and raised our confusion, and BUCT-China's students looked at our plasmid map and found that there were enzyme cleavage sites on both sides of the receptor. BUCT-China told us that we could try to construct the plasmid by using the enzyme linkage. The plasmid was constructed by linking the promoter to the fluorescent protein, and then the receptor was inserted.
After two more failed experiments, we succeeded in constructing a bitter plasmid in the third enzyme-linked experiment.
For the construction of umami flavor plasmid, we designed it by what we learned and planned to use the same enzyme cleavage enzyme linkage for the construction of plasmid, but due to the competition time, we were not able to successfully construct it within the time limit, so in the next time, we will continue to try to finish the construction of fresh flavor plasmid.
According to some communication tips from everyone in the mid-term, two teams shared for their modeling solutions and exchanged opinions with each other
The suggestions we received from BUCT-China:
The students of BUCT-China team suggested that the yeast cell growth model used by BIT-China team was a good idea and wanted to know more specific things.The students of BIT-China team pointed out in detail that the yeast cell growth model was established mainly to simulate the growth of yeast cells and fine-tune it according to the real experimental situation and fine-tune it, and this model relies mainly on focusing on the changes in the concentration of individual transducing substances.
The suggestions we provided to BUCT-China:
The BIT-China team proposed that the "artificial meat skeleton" model of the BUCT-China team has high application value. According to the different structure of the skeleton, cross angle, porosity, and diameter of the skeleton, the computer simulation can determine the conditions under which the meat can grow better, thus saving more time for the experiment.
At the same time, both teams had their first prototype website pages and contents, and students from the artwork groups of both teams exchanged ideas about their respective designs and suggested improvements to each other.
We first put forward our views on the expression packs made by the BIT-China team, and after that, we discussed and made a small plan for the two teams to draw each other's materials for the artwork part, for example: small yeast with the project gradually grows up, from childhood to youth; artificial meat,from rough to fine to beautiful a process.
On September 30, 2021, BIT-China team and BUCT-China team cooperated to conduct a popular science lecture with the theme of "ethics". BIT-China team talked about the ethics of artificial life from the part of "biosensor" in the 2021 project, while BUCT-China team talked about the ethics of "cell culture meat - artificial meat" design from the project "artificial meat".
The BIT-China team gave the audience a deeper understanding of our project through an interactive session on site, while the BUCT-China team communicated with the audience through the online format.
Expectations for future cooperation
During the whole season of cooperation, everyone's projects have been improved a lot, and the team members have gained more new ideas and creativity from the communication.
In October, we had a wrap-up online meeting with BUCT-China (online as well, if the situation allows). During the meeting, we summarized our respective projects, as well as our thoughts and reflections on our partnership throughout the season, and our vision for the future. At the end, we all took a group photo together, leaving precious memories and contributing to our deep friendship.
This year, our partnership with UCAS-China is multi-faceted and multi-faceted. The UCAS-China team's project is to synthesize two enzymes, Cdh and CkTcS, from E. coli, and immobilize them in the cartridge of a special cup, which will convert caffeine into bitter theophylline for personalized caffeine intake management. Our project hopes to obtain various flavored peptides by enzymatic digestion of soy protein, and then use the combination of different peptides to develop flavorings with different tastes, so as to remedy the problem that some expectations of the public for flavorings in the current market are difficult to be met.
Because the projects of both teams are related to food, we discussed and decided to collect some standards about food safety evaluation first, in this regard, we got the safety evaluation standard of UCAS-China team about food enzyme preparation, which provided us with the standard for peptide testing afterwards. At the same time the students from UCAS-China had some discussions with researcher Tao Yong from Institute of Microbiology, mainly about the current market acceptance of synthetic food, which also has some significance for the future prognosis of our seasoning.
Practical Research Cooperation
And, given that there will be some fine-tuning of the project direction, we released the relevant questionnaire and the two teams shared the results of the questionnaire research with each other. We also learned that the questionnaire results obtained by the UCAS-China team showed that the public does not know much about the effects of caffeine on the human body and the caffeine content in various beverages, and many respondents also expressed interest in coffee-related science, so they also plan to design the app as a science platform for this project, using coffee advice, science articles and health tips We plan to increase the usage scenarios of the app. At the same time, we had already designed the Flavor card app at the beginning of the project, which was also part of the subsequent communication between the two teams.
In terms of experimental part, the initial goal of the UCAS-China team students was to decaffeinate the coffee without destroying the flavor, but there was a lack of experiments to verify the effect of converting caffeine to bitter theophylline on the flavor, and some coffee lovers, after learning about the UCAS-China team's project, also suggested that unique flavored specialty coffees could be produced. So there was some exchange on the aspect of flavor that people like.
At the same time, because our project this year focuses on some innovation and breakthrough in flavor detection, according to our previous knowledge, the current biological quantification of flavor is almost blank. Therefore, we designed human-derived taste receptor heterologous expression of Saccharomyces cerevisiae as a taste detection officer, and used fluorescence intensity to quantify taste by modifying MAPK pathway to amplify the signal. We plan to use our biosensor to help the students of UCAS-China team to detect the degree of coffee bitterness and give the results back to the students of UCAS-China team to help them to make better specialty coffee.
Meet the Unique Taste
In terms of applet creation, the exchange focused on the design of questions and related contents. The main way was that the two teams helped each other to promote the questionnaire, so that the applet could be created based on the hot topics collected from the questionnaire.
The UCAS-China team's design of the app was divided into three modules: collection, analysis and Bluetooth transmission. The collection module asks users to fill in their height, weight and eating habits when they first log in for subsequent analysis, and they can also record their daily coffee intake, intake time and sleep, as well as their heart rate and smoking history.
This is similar to our Flavor Card app, where the dishes are divided into two categories: vegetarian and non-vegetarian. In the website customers choose the dishes they want to try in the menu according to their mood or thoughts at that time, and also choose their favorite level, and certain records will be made in the background.
In the analysis module, the students of the UCAS-China team built a model through the results of a population cohort study on the effects of caffeine on people, which can give recommendations on the upper limit of caffeine intake. At the same time, they used deep learning algorithms, combined with individual user data to make the model more personalized, in line with the large individual differences in the effects of caffeine on people. More interestingly, the applet designed by the UCAS-China team is connected to the hardware via Bluetooth and is able to detect the temperature of the drink and the caffeine content, so that users can use our enzymes under optimal conditions and record their caffeine intake.
Our backend Flavor Card applet processes the data based on the customer's choice to get a numerical value of the customer's preference for the five basic flavors, and creates a report with unique colors and flavors for feedback to the customer, and then our "exclusive flavoring" will be based on the data in the Flavor Card to The Flavor Card is used to customize the flavor profile. At the same time, our Flavor Card website can collect people's favorite flavors from all over the world and compile them into a flavor database, which is a useful reference for food companies.
Build iGEM Online
At the same time, in order to better train iGEM newcomers, UCAS-China team members plan to develop an online catechism teaching platform iGEM EduHub, inviting schools to share their training materials for resource sharing and win-win cooperation, thus achieving high quality and high efficiency. Moreover, the UCAS-China team has made a preliminary plan for this platform, which will mainly consist of two parts: courses and forums, and a course classification system for all aspects of iGEM has been built.
At the same time, in view of the characteristics of the online teaching platform, the students of the two teams discussed and proposed a plan for future team recruitment, perhaps by first training the students and then selecting and forming the team, which would be of great help to the team's subsequent schedule progress.
Through the cooperation with the students of UCAS-China team, our project has been improved in many aspects. Students from both teams exchanged inspirations with each other in the cooperation, and both of them got improved. Meanwhile, I hope we can have more cooperation in the subsequent competitions and work together to improve!