Our team is willing to maintain meaningful relationships and carrying out in-depth cooperation with other iGEMers who have similar motivations.We believe that iGEM is not only a competition but also a golden opportunity to encounter and engage with students with similar passion of Synthetic biology. This year, we found two excellent partners--the team of ASTWS-China and Fudan.
Right at the start of this seasons' iGEM competition, we heard from that the team of ASTWS-China is interested in the project of our past team carried in 2016 which focused on the surface display separately using E.coli and Yeast and mutant its genes to modify PET hydrolase(PETase) . They read wiki the team of 2016 TJULSL-China and emailed us, asking whether they could get some information of their project. Coincidentally, this year, our team is also committed to environmental issues caused by PET. They expressed their will of discussing how to design projects and do some background researches together with us. We responded them as soon as possible. After that, we held several rounds of online meetings to discuss PET background and current solutions and so on through which we surprisingly found that both of us are devoted to improving the efficiency of PETase to degrade PET. In the early meeting, we exchanged ideas of each others and reach an agreement that PETase has a low degradation efficiency.
Through the following month, we approached each other by twice online meetings to further discuss the certain strategy. In the project cooperation, we have an active discussion with the team of ASTWS-China in always online, and we often give constructive suggestions to their projects. For the first time, our team members had a brainstorming and list all the solution we learnt from papers. We finally reached a consensus that Improve the contact efficiency of substrate and enzyme. For the second time, we reminded them the degradation rate of PETase was low at normal temperature, which led to their further thinking and project improvement.`
During this period, we held six meetings online. In the first five meetings, we trained them on molecular biology, and in the last meeting, we exchanged background research information.
For the first time, In the first meeting, we introduced the physical and chemical properties of nucleic acid, including its strong ultraviolet absorption, the role of ATP in energy metabolism, and the renaturation of deformed nucleic acid.
For the first time, we introduced molecular cloning methods, including the role of the restriction endonucleases, a variety of PCR methods, and some methods of expressing cloned genes. In this meeting, we were surprised at their depth of understanding of biology, and we realized Improvement of high school students’ biological knowledge by iGEM.
For the second time, we introduce molecular separation methods including gel electrophoresis, two-dimensional gel electrophoresis, in exchange chromatography, gel filtration chromatography, and affinity chromatography. During this meeting, they asked some questions which is also a review and improvement for ourselves.
For the third time, we introduce some application of nucleic acid hybridization, including southern blot, in situ hybridization, western blot and other technology. In this exchange, they once again showed their amazing basic knowledge and had their own opinions and understanding about our introductions.
For the fourth time, we present some tracer techniques including autoradiography, phosphor screen imaging, liquid scintillation counters, and non-radioactive tracer techniques. They were very interested in some of these technologies and had additional communication with us.
For the fifth time, we introduce some techniques related to DNA sequencing, including Sanger chain termination sequencing, automated DNA measurement, and high throughput measurement. This is our last lesson to share. We have extended the part outside the course and we all enjoyed the exchange very much.
For the sixth time, we introduce some techniques related to DNA sequencing, including Sanger chain termination sequencing, automated DNA measurement, and high throughput measurement. This is our last lesson to share. We have extended the part outside the course and we all enjoyed the exchange very much.
For the last meeting of this period, we conducted background research and information exchange, which saved both parties’ time and enabled both parties to have a deeper understanding of the current situation of PET degradation.
At this stage, the team of ASTWS-China conducted a wet experiment, and found that there was still room for improvement through the experimental results. We initially planned to measure the concentration of MHET （biodegradation product） by HPLC to test the efficiency of our PET biodegradation system. However, the machine broke down accidently. The team of ASTWS-China discussed this problem with us immediately. We worked together and stayed up for nights and solved this problem efficiently by hydrolysing para-Nitrophenylbutyrate (pNPB) ( hydropysis properties similar to PET) and measuring OD of its hydrolysis product p-nitrophenol, finally getting the result by a series of calculations.（This method is known as pNP-Assay）
After successfully measuring the data as expected, the team of ASTWS-China tries to build model but they meet some problems. We use the software called Metlab and obtain parameters in their equation. Throughout the process, we helped them a lot. Also, we introduced the related knowledge of Michaelis-Menten equation and how to apply it in application in enzymatic reaction.
Also, the experimental experience of the team of ASTWS-China is meaningful to prevent our experimental problems in the coming year. Their problems in the experiment can be used as experience to help us avoid risks and reduce the possibility of failure in the experiment.
From September to October, our experiment came to an end. We were working day and night for iGEM and encouraging and supporting each other to spend the last time. We were all actively preparing the content of the competition. We exchanged PPT contents and encouraged each other. We gave advice to each other and constantly improved ourselves. For example, we once had a heated discussion on the background part, and constantly revised it according to each others’ suggestions.
As we all know, as the COVID-19 epidemic has swept the world, the normal operation of all countries in the world has been affected and it is difficult to work normally. Although the situation is very difficult, we insist on working and actively promote the project. The team of ASTWS-China together with us encouraged and supported each other and passed this difficult period together.
With team Fudan
In August 2020, we got in touch with Fudan, hoping to reach a cooperation with them. After discussing the projects of this two teams this year, we decided to hold a summer camp for junior and senior high school students on line, mainly using the app Tencent Meeting. During the summer camp lasting two days, students from our team explained the knowledge about synthetic biology and our iGEM project, expecting to stimulate their interest in biology, synthetic biology and iGEM. Meanwhile, we invited Yaxin Wang, the lecturer of School of Life Sciences in TJU, to give a academic lecture on COVID-2019, which was expacted to have them understand the knowledge of virus.
Thanks a lot to our cooperation and communication and the summer camp we held, we have realized that the biodegradation of plastics is actually not well known by many people, and the public does not have enough consideration on plastic pollution. At the same time, our recognization on the importance of our projects to the environment has been deepened. Therefore, we will put much more energy into our project.