Team:ASTWS-China/Partnership
Partnership
Welcome to iGEM 2021
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Partnership with IvyMaker-China
Overview: Our team and IvyMaker-China have had virtual meetups every 1-3 weeks from April to have discussion about progress, problems we met in the framework of wet lab, human practices, education, and model etc. Through this process, we have become friendly and mutually partners.
TIMELINE
1 Wet Lab In each meeting, we shared our progress of the wet lab, discuss problems and provide help and suggestions. Besides, taking account of the different expression systems of our projects, we have been working hard to facilitate the mutual verification of the prokaryotic and the eukaryotic expression system, such as enzyme expression efficiency, enzyme activity, etc. 1. We planned to use plastic fragments as the substrate, but the response was not good. IvyMaker-China shipped their PET powder to us so that we could continue the experiment.
2. Using p-NP assay, we determined and compared the enzyme activity of PETase and the impact of the addition of MHETase. The detailed results and conclusion are as followed. Conclusion: Compared with the data obtained by two teams independently, it is found that the addition of MHETase could help to improve the enzymatic activity of PETase to some extent. ASTWS-China
We selected p-Nitrophenylbutyrate (p-NPB) as the substrate, which can be hydrolyzed by PETase to p-nitrophenol (p-NP). Using p-NP assay, the enzyme activity of PETase and the impact of the addition of MHETase can be characterized by measuring the optimal optical density (OD) values at 405 nm.
Figure OD405 of p-NPB (0, 0.5, 1, 2, 4, 8 mM) hydrolysisby overexpressed PETsae-SC+OmpR and PETase-SC + OmpR + MHETase-ST, respectively. IvyMaker-China
To determine the Michaelis constant of PETase, select p-nitrophenyl acetate as the substrate; at the same time to explore whether the addition of MHETase will affect the kinetic parameters of PETase.
Figure Determination of kinetic parameters, fitted curves of the Michaelis equation of PETase (left) and MHETase (right). 3.3. We were also planning to use western blot to determine the protein expression corresponding to a certain cell concentration and compared the data.
2 Human Practices 2.1 Background research With IvyMaker-China, we made four lists as our own research reference and also for the good of future iGEM teams if needed, which you could find detailed info in CONTRIBUTION page. 1.manual of different medias for live broadcasting 2.a list of some previous projects using PETase and MHETase 3.a list of research paper for reference 4.a list of laws/ regulations/ policies about plastic recycling in the process of collection, production and such.
2.2 HP activities We share feedback on our HP activities in monthly virtual gatherings for mutual reference to further improve the project. For example, we shared the experience of interviews with local sewage plants, and IvyMaker-China shared their feedback on participating in CCiC8.
3 Model We had several meetups with IvyMaker-China to explore the potential of the model design and the problems we met. IvyMaker-China only did one GPI-anchor protein prediction network model at the beginning. While we later proposed them to try the Michaelis equation and considered using MHETase as the cofactor of PETase. This later became their second model. In the progress, we continued to discuss with each other, discussing problems, the modeling method, and the results we obtained.
4 Education We invited IvyMaker-China to share their projects on our platform. We launched an online public education and communication lecture called "GENE HACKER" with IvyMaker-China, Worldshaper-Nanjing and Worldshaper-Shanghai. We invited highschoolers and their familes interested in synthetic biology or bio in genreal to join this event and shared background info and lab design of our projects.
Partnership with ZJUT-China
The communication between our team and ZJUT-China started in July, kept till nowadays.Our conversations focused on works of Human Practice.
08.29 Online Meetup We hosted this online meeting to enhance communication between HP and Dry group hardware with each teams. ZJUT-China modified the event plan provided by us before the conference. The members of the two teams had in-depth discussions on the merits of the scheme, and finally came to the optimal scheme, which ensured the smooth progress of meetup. This activity not only allowed the HP group and dry group of the participating teams to fully communicate, but also provided valuable experience for our team to open online Meetup.
09.28 Qiushan Primary school In order to bring the concept of "biology" to the young people in China, ZJUT-China cooperated with Qiushan Primary School to bring an interesting biology lesson to the fourth grade children in the form of PPT and handwork by four team members. We communicated with ZJUT-China, and eventually found an education method suitable for the younger age group. It has been proven that our education methods for the younger age group are effective. The exchange between the two teams promoted the development of the activity and enriched our educational experience for the young population.
10.18 An information session at the highschool In order to promote iGEM and our own projects, we held an information session at the school on October 18th. Since ZJUT-China's academic forum is similar to this activity, they shared their PPT and lecture content with us. Meanwhile, they exchanged our experience in carrying out the lecture and provided help to them. This event enhanced the reputation of iGEM and ASTWS-China and attracted more peers to pay attention to synthetic biology.
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