Team:ASTWS-China/Partnership

Preface

Inter-teams cooperation is always encouraged by iGEM which gave us an unique experience during a competition. We felt that iGEMers are not competitors like participators of most of other competitions or contest but partners to achieve the goal together. This is an excellent experience as well as an important lesson which taught us that “win-win cooperation” is much more valuable than “zero-sum competition” when we are facing with problems and challenges.

Very luckily, besides cooperation with other high school and university teams as we mentioned at cooperation part, we had a great partnership with our partner - TJULS China from Tianjin University. During the whole iGEM season 2020 even from mid-march, faced our common interests and goals on plastic degradation by PETase, we worked physically individually but mentally together, we helped each other, encouraged each other and supported each other.

To be honest, due to differences of age, professional knowledge, academic ability and available extracurricular time, we received more help and assistance from Tianjin University team than what we might offer. We appreciate for their efforts and are proud of our contributions as well.

Co-op Activities

1. Background research and brainstorming

Our project was inspired partly by the project done by TJULS China 2016（Gold Medal，Best environment project）. That year they also aimed to degrade PET. This project was to mutate its gene sequence and make surface display separately using E.coli and Yeast. After reading their wiki, we emailed them and were wondering whether we could discuss how to design our projects and do some background researches together. Surprisingly, we received their reply very soon and got to know that they were also focusing on the environmental issues from PET by lucky coincidence.

After that, we held several rounds of online meetings to discuss PET background and current solutions. Because of COVID-19 outbreak, everyone had to stay at home and none of team members from both team may meet in person. Conducted background research and information exchange saved time and help better understand the current situation of PET degradation for both of two teams. These continuous online discussions and brainstorming benefited both of us to find our project directions. We both agreed that PETase without strengthening has low biodegradation efficiency at room temperature. Therefore, we decided to work together to improve PETase but on two dimensions: making it work at higher temperature and strengthening PETase produced by E.coli-biofilm system.

2.Training and discussion

During our training period, we regularly held on-line meeting with TJULS-China. They offered lots of papers and lecture materials on molecular biology, experimental skills as well as modeling to us and taught us a lot, such as nucleic acid, PCR, gel electrophoresis, two-dimensional gel electrophoresis, in exchange chromatography, gel filtration chromatography, affinity chromatography, southern blot, in situ hybridization, western blot, etc. Both of believed the training process, including Q&A and discussions, helped not only us (as students) but also them (as teachers) to better understand corresponding knowledge. The figures below are screenshot of these on-line meetings.

3. Experiments

Because of strictly COVID-19 epidemic prevention and control requirements, TJULS-China could not go back to lab and do experiments at early summer. Our team ASTWS-China luckily got a precious chance to get into the lab in Hangzhou for July and August. We have invited them to shared lab and do experiments together. Although because of complex reasons including epidemic situation, safety, financial support, etc., they could finally not come to Hangzhou and join us, we truly want to help each other and progress together.

Furthermore, when we were doing experiments, we concluded some useful experiences. For example, we found that weighing the mass change of PET with high-precision machine was unfeasible and should not waste too much time on it, especially the lab time is very limit for every team this year. Based on the experiences, we shared also our valuable estimation and suggestion towards certain experiments with TJULS-China, which also helps them.

4. Modeling

As high school students, due to the lacking knowledge on modeling and mathematics, even we have taken a short term courses, modeling was always the most challenge part for us. Fortunately, TJULS-China helped us a lot on this part throughout the whole period.

We initially planned to measure the concentration of MHET（biodegradation product of PET）by HPLC to test the efficiency of our PET biodegradation system, and use this result for enzyme-catalysed reactions model - Michaelis-Menten equation. However, the HPLC machine broke down accidently. We discussed this problem with TJULS-China immediately. We worked together and stayed up for nights and solved this problem efficiently by hydrolysing para-Nitrophenylbutyrate (pNPB) (hydrolysis properties similar to PET) and measuring OD of its hydrolysis product p-nitrophenol, finally getting the result by series of calculations.（This method is known as pNP-Assay.）After successfully measuring the data as expected, they help us use Matlab to model, calculate and obtain parameters in our equation.

5. Final presentation preparation

During the last two weeks in October, both of us were exporting all our energies on wiki, final video presentation and poster. We exchanged PPT contents, helped double check results and English expression, gave advice and suggestion, encouraged each other. These advices and suggestions help both of us improve ourselves.

Ending

Because of COVID-19, 2020 is a really tough year for all over world including our iGEMers. Unfortunately, we had to give up or change many our plans for the project, lost the opportunity to attend the Giant Jamboree and meet iGEMers all over the world in Boston as we had expected. Fortunately we found our great partner TJULS China. These unique experiences that we shared is unforgettable for both of us. Thanks for everything! We send all of our best wishes to TJULS China team for a wonderful result during Virtual Giant Jamboree and all success possible to everyone in the future.