1.Partnership with NJTech_China

Overview

We work with NJTech_China team has been cooperating from the initial brainstorming to the final publicity activities.

May&June ：topic brainstorming&background survey

July&August&September ：experiment&Theme meetup

September&October ：writing relevant documents

1.Brainstorm and discuss topics

Our two teams shared the background information about each project online. Our research directions are similar, so we can learn from each other and share the survey data.

2.Laboratory cooperation in summer

Our team and the students of NJTech_China team completed the project tasks in the same laboratory this summer, solved the problems encountered by each team, and actively helped each other. The NJTech_China team provides suggestions on the design of experimental ideas for our senior high school team, shares experimental materials (including strains, plasmids or sequences), and guides experimental teaching.

(1)Guidance of experiment ideas and training of experiment operation process

They helped us analyze problems in the PCR process, optimize the PCR reaction system, and enable us to successfully amplify the fragments

(2) Provide experimental biomaterials

- The plasmid Pthl 82151 allows our team to amplify

- Provide genome of Urumqi bacteria, and we amplify specific dps genes in the bacteria.

in order to enhance Clostridium tyrobutyricum antioxidant ability, we have introduced the gene fragment dps extracted from Urumqi bacteria by Nanjing Tech University. The significant antioxidant function can make Clostridium tyrobutyricum adapt to the aerobic environment better. we also learn from their amplification methods to amplify the dps gene fragment

（3）We used the thl promoter, and the results obtained by polyacrylamide gel electrophoresis experiment verification and growth performance measurement were not ideal, so NJTech_ China suggests that we replace the thl promoter with the plac promoter provided by them.That’s proved effective in our further experiments. And because of this,our team provides the idea of optimizing the thl constitutive promoter to urge the team of Nanjing Tech University to continue to study and upgrade this promoter.

(4)However, improving the promoter has always been a challenging desire for us, which thoughts of temptations made us curious. After consulting literatures and in-depth exchanges on this ideas with team NJTech_China, they suggested we try Pvgb, which has a certain induction effect of oxygen to implement the promoter engineering strategy, because the improvement of the PthlA promoter is considered more difficult for us.

From NJTech_China’s introduction on promoter vgb, we learnt that it’s a microaerobic induced promoter from Vitreoscilla hemoglobin gene with an initial FNR(fumarate-nitrate-regulator) binding site in its sequence.

Trying to enhance the regulatory effect of FNR on vgb promoter, we decided to insert another FNR binding site(which sequence was found on a research paper as TTGATnnnnATCAA) upstream the native binding site, so it’s expected that the effect of regulation by oxygen concentration is more sensitive than before.

By expressing the flavin mononucleotide based fluorescent protein Bs2 provided by Dr. Zhengming Zhu’s research group, the fluorescence intensity observed could be used as a reference to determine whether the modified promoter meets the expected effects in different oxygen concentrations.

3.Education meetup

We attended the education activity exchange meeting held by the college student team to learn how to carry out such activities，which gave us a chance to learn from other teams how to run theme events.It was impressive that the team NNU_China held a wide range of theme activities, especially for the younger age groups, that were both unique and relevant

2 Partnership with Worldshaper-NJBIOX

Overview

Worldshaper-NJBIOX and our team carried out project implementation in the same laboratory. Since the establishment of the team in March, the two teams have been preparing for the competition together with the relationship of partners. They have maintained close communication and cooperation in the early stage of theoretical knowledge learning, topic brainstorming, background research, experimental skills learning, project experiment implementation, public education activities, modeling work and hardware design.

1. Brainstorm the topic

In the early stage of topic preparation, our two teams carried out brainstorming discussion, and we made a survey and discussion on the background of Clostridium tyrobutyricum.

2. Wet lab

2.1 Experimental skill learning

We have jointly participated in the summer training camp of NJTech_China, exchanging experience in learning

2.2 Experiment

The Worldshaper-NJBIOX team helped our team analyze the problems in the PCR process, optimized the PCR reaction system, and successfully amplified the fragments.

The Worldshaper-NJBIOX team helped our team analyze the problems in the PCR process, optimized the PCR reaction system, and successfully amplified the fragments.

Technical problems during part registration:Worldshaper-NJBIOX helped us extract pmtl-82151 vector plasmid, and configure the reagent for nucleic acid electrophoresis

3. Public activities and meetups

3.1 Publicity activities

During the summer, we plan to organize an online promotion to introduce our topic and this competition iGEM to everyone while we experiment. Our team has members who are good at art and design, so we were in charge of designing the posters and Worldshaper-NJBIOX was in charge of planning and hosting, and, they invited the Asian ambassador of iGEM 2022, to participate.

On October 5th, we conducted an offline promotion activity in Lanyuan community in order to reach out to the people. In this event, we jointly produced comic booklets for publicity and promotion.

3.2 comic manuals

We also worked together to produce a comic manuals for promotional purposes. We design the layout and content of the manuals, while Worldshaper-NJBIOX reviews and funds the printing.

3.3 Project meetups

In order to learn more about other teams' topics and to broaden our minds and eyes, we co-organized an offline topic exchange meeting on 9.29 and invited two other high school teams. In addition, we also participated in the project exchange and education exchange held by other teams.

4.Exploration of modeling ideas

In July, we started preparing to build the mathematical model. And in the meantime, we worked with Worldshaper-NJBIOX to discuss how to build it. We provided them with ideas and directions, i.e. analyze the mode of butanol and butyric acid production, to help them build the current model.

5.Hardware design improvement

The team Worldshaper-NJBIOX designed the hardwares, in which they encountered difficulties in adjusting the speed of the crushing part, and our team provided advice on adjusting the speed, which helped them solve the problem.