Section 1 : Team Introduction
Team Tsinghua comes from Tsinghua University in Beijing, China, and is a team of students from the School of Life Science, School of Medicine, as well as the School of Arts and the School of Aviation; Tsinghua has designed many excellent projects since its participation in 2007, taking advantage of the University's strengths in the fields of cell biology and biochemistry.This year's Tsinghua made an exploration in the field of reproductive health, hoping to develop a sperm quality detection chip for home use and contribute to the current situation of declining male reproductive health, which is becoming increasingly serious. Tsinghua University also involved the Pmr system in the wetlab design, and has a series of collaborations with HUST around this.
Section 2 : What Tsinghua brings to HUST
We have communicated with many teams this year. During the communication with Tsinghua, we found that we all have chosen pmr two-component system as a part of iGEM project this year, and have made similar modifications, namely peptide replacement of membrane protein pmrB. The GolS operon in our project is also similar to nisin, another Tsinghua expression pathway. Therefore, we immediately set up an online meeting and had many conversations.
Fig. 1. Meeting 1
In the process of communication, we found that the problem of low expression efficiency of pmrC promoter encountered by Tsinghua in the construction of Pmr two-component system was exactly similar to that encountered by us. We started talking to each other about our plasmid map experiences right away.
After we had obtained some experimental basis, we exchanged plasmids with Tsinghua and asked them to verify whether the GolS system could successfully and persistently anchor the target protein to the membrane, and achieved good results
Fig. 2. Results from Tsinghua University
In terms of dry laboratory, we have noticed that Tsinghua's wiki optimization in past years is very excellent, and we have consulted them for their experience
Fig. 3. A souvenir of the exchange of plasmids
In the end, the painters of the two teams drew together to record our partnership
Fig. 4. THU-HUST
Section 3 : What HUST brings to Tsinghua
Our relationship with HUST stems from the two-component system we both chose to use, Pmr, which is highly versatile and scalable and can be applied to a variety of protein detection scenarios. After learning about each other's project designs, we quickly contacted HUST and conducted our first online meeting to introduce our projects to each other. We also asked HUST questions related to the timing of induction and expression of the Pmr system, helping us to troubleshoot some possible problems during the experiment.
As we encountered the problem of poor system stability in the construction of the Pmr two-component system, we exchanged with HUST the experiments designed in the strength validation of the two-component system, and asked HUST to obtain the map of their plasmid constructs and compare them with our plasmid constructs to find if there was a problem in the design of the primary sequence of the receptor.
Around September, we completed multiple induction experiments with both our Pmr and nisin systems. To verify the stability of the system, we asked HUST to complete two nisin induction experiments for us to investigate whether the nisin system could maintain its effectiveness even under different contexts. After we sent the relevant plasmids and reagents by courier to lab of HUST, they quickly completed the two induction experiments for us and obtained similar results as we did in our own lab (link to result). We benefited greatly from this mutual assistance in wet experiments.
Fig. 5. Online meeting discussing on our experiment result
It is worth mentioning that during our pre-experimental stage, we had conducted a project on laccase. We found out that HUST had used laccase components in previous projects, so we contacted HUST and asked them to send us the components at that time, which allowed us to conduct the experiments quickly. Although this project did not turn out to be our final project, we are still very grateful to HUST for their generosity, and perhaps this was the beginning of the relationship between our two teams.
Fig. 6. A souvenir from HUST
