Team:NWU-CHINA-A/Partnership
SZPT-CHINA
https://2020.igem.org/Team:SZPT-CHINA/PartnershipDuring the 4th Southern China Regional Online Meeting, our two teams had a general understanding of each other's projects. Their project is about the prevention of dental caries by killing Streptococcus mutans. They expressed a polymer that can be hydrolyzed by salivary hydrolase to monomer Tachyplesin. We found that both of our teams used antimicrobial peptides and there was an overlap in the screening of antimicrobial peptides, which we used as a breakthrough to carry out a number of exchanges and cooperation.
The student leader of the two teams communicated with each other. At first, SZPT-CHINA wanted to use tooth socket as drug carriers to design hardware, but our team felt that braces had shortcomings such as high cost and individual needs to be customized, so we suggested that they could use some more convenient, faster and lower-cost products, such as toothpaste.
Shaanxi iGEM Meetup is the first communication meeting we hosted.
We specially invited members of our partner SZPT-CHINA to participate in Shaanxi iGEM Meetup and had friendly exchanges with all teams in Shaanxi. After introducing our project, we both had a detailed understanding of each other's projects. They were interested in the broad-spectrum antimicrobial peptides we used.
Our team was invited to join the Friend Team Meetup hosted by SZPT-CHINA. In that meetup, we communicated about antimicrobial peptides. We exchanged problems and experience about the selection of antimicrobial peptides and the design of experiments. After that, we got a lot of valuable suggestions.
We communicated with each other online, and we provided them with a variety of relevant antimicrobial peptide sequences to help them get the minimum inhibitory concentration of multiple antimicrobial peptides. Finally, they chose to add DCD1L antimicrobial peptides into their experiment.
In this online meetup, we mentioned that we lack a suicide switch that prevents bacteria from escaping, and they suggested that a nutrition-hungry suicide switch might meet our needs, and would like to provide us with a mature suicide switch that they used last year. The suicide switch can sense the concentration of glucose in the environment. Glucose starvation activates the catabolic response using cyclic-AMP (cAMP) receptor protein (CRP), one of the best-studied TF’s. Low glucose environments cause the cAMP-CRP complex to bind target DNA and regulate hundreds of gene targets. We used PT-α crp to control the downstream gene. It is sensitive to glucose starvation. When glucose is deficient in the environment, glucose starvation induces the overexpression of autolysin gene in the T-α crp promoter(acmA).
We communicated with each other about the progress of the experiment. As a result, we exchanged our protein purification method because we all encountered the problem that the protein could not be purified and the concentration of the purified protein was too low. They recommended us to use the ultrafiltration centrifuge tube for protein purification and concentration. When we took their advice, we made some progress in our experiment.
SZPT-CHINA team and we determined to hold a lecture. After discussion, the main direction is developing and implementing synthetic biology. The purpose was to introduce synthetic biology development history, the basic knowledge of synthetic. Besides, we would like to focus on the antibacterial peptide and how to produce antimicrobial peptides by means of synthetic biology.
Our two teams held a joint lecture and carried out modeling communication successfully. At last, we also designed the cartoon characters of cooperation between the two teams for promotion.
Beijing_4ELEVEN
https://2020.igem.org/Team:Beijing_4ELEVEN/PartnershipWhen Beijing_4ELEVEN completed our questionnaire, they found that our projects have something in common.
The projects of our two teams have one thing in common.
We all hoped to solve the problem of antibiotic resistance by using antimicrobial peptides, and we both used materials that can stick to the surface of the skin. We noticed there was a great potential for cooperation.
We see great potential for cooperation.
Our two teams were arranged in the same ZOOM conference room in the 4th Southern China Regional Online Meeting(CCiC). We learned that their project was to obtain a product that could form a waterproof membrane on the skin surface by using Pichia pastoralis to express sticky protein and tyrosinaseantimicrobial peptides which can kill propionic acid bacteria and express. The product helps prevent acne and can be easily removed by protease hydrolysis, avoiding the pain of direct skin tearing.
After the free discussion of the report, we found that we both had the willingness to cooperate with each other. Then the team leader of Beijing_4ELEVEN sent us an email for cooperation request. We were willing to receive the request.
When we had our first formal online communication, we discussed the knowledge about antimicrobial peptides in the Tencent meeting. And we communicated with each other about the antimicrobial peptides we used. Beijing_4ELEVEN shared with us their experimental data related to LL-37 human antimicrobial peptides. We really appreciate that. The figure below is their data. The protein sequence of human cathelicidin LL-37:LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTE
Antimicrobial peptides only affect the area in direct contact on E.coli1655
We also discussed how to advance HP in the future.
Team members from both sides set up WeChat group chat, so as to discuss relevant issues later.
On the same day, we communicated about the progress of the experiment. We asked the teacher about the relevant experience of protein expression and purification experiment, and we improved our experimental design.
Beijing_4ELEVEN and NWU-CHINA-A described the general content of the project and the main modeling direction respectively, discussed the problems in the modeling of each other, learned from each other, and optimized the modeling details of each other.
In this communication meeting, Beijing_4ELEVEN found that bacterial mucin may be of great significance to our project, so they suggested us to look for Greatbay_SCIE2019 project. Then we checked the literature of bacterial mucin and supplemented the project.
Beijing_4ELEVEN shared their experience on the Bronze Contribution, which helped us a lot, and we adjusted the direction of the experiment accordingly. We shared with their team about the method of PHA membrane formation.
We discussed the problems with our respective wikis and check the content of each part.
XJTU-China
https://2020.igem.org/Team:XJTU-China/PartnershipXJTU-China and NWU-CHINA-A conducted online communication activities. The two teams have been partners of iGEM for many years. This time we will continue to cooperate. In the exchange meeting, we exchanged each other's recent ideas and the two teams brainstormed together.
We invited XJTU-China to participate in the Shaanxi iGEM Meetup hosted by us. We shared each other's projects and gave suggestions in the meetup. We also have extensive exchanges and cooperation in experimental methods, websit econstruction and parts. In terms of experimental methods, we exchanged some matters that should be paid attention to when cultivating strains, and summarized the experience and reasons of their failure. XJTU-China used GitHub and other methods to compile wikis, and established perfect webpage wikis, which inspired us greatly. Therefore, under the cooperation of the two groups, we successfully established our own website wikis.
The student leader of XJTU-China proposed to supplement the Part library. The Northwest Part library is a gene library jointly constructed by Shaanxi. Before each iGEM competition, the Shaanxi teams will upload the genes they think are interesting to the library. Both teams are actively preparing for the construction of the Northwest Part library.
Our team has reached cooperation with the XJTU-China team in modeling. Our modelers helped find the parameter data needed by each other's growth and expression equations. We reached a consensus on the application of some formulas, discussed the CA model in depth, and exchanged modeling methods and experience. The XJTU-China team has encountered many problems. The parameters needed during the bottleneck period are difficult to find in the literature. After clarifying the related problems, our team pointed out that it should be combined with experimental data for parameter fitting. At the same time, XJTU-China also solved some of our problems. Our growth model lacks some parameters. The suggestions they gave us revolved around programming and GAN, and we exchanged the PH and soil parameter measurement experience. We are all aware of the importance of querying data, and we are more clear about the direction of modeling.
We conducted experimental exchanges, in the course of their experiment, they found that the negative control had a band, we suggested that they use new water, or used new primers, and the primers were packed into small packages. We had trouble in lysing cells, and they suggested that we replace the Bacterial lysis solution. Before: Bacterial lysis solution 1.0: 50mM/L tris-HCl 0.5mM /L EDTA 50mM/L NaCl pH=8.0After: Bacterial lysis solution 2.0: 20mM/L tris-HCl 150mM/L NaCl., after changing the bacterial lysis solution, the efficiency of breaking bacteria was improved a lot.
We had a modeling exchange meeting once again. NWU-CHINA-A suggested XJTU-China that they can first fit the corresponding parameters, then select the appropriate promoter from the part library to construct the system and optimize the experimental design of the project with dry simulation to solve problem that the parameters of the suicide switch activation mechanism cannot be found. XJTU-China modeling members made suggestions for the simulation process of cell expression products in the NWU-CHINA-A's modeling.They advised that probability distribution can be used to replace the assumption of uniformity of all cells that make the simulation more realistic and credible.
Both teams checked and discussed this year's award criteria, and under the change of this year’s award criteria and the obstruction of the epidemic, we agreed on what should be more important. Both sides had benefited a lot, which is the strength of Shaanxi iGEM team alliance.
Finally, our team decided to put Kaia, KaiB and RPAA genes into the part library, and we will use them later.
The order of NWU-CHINA-A team in IDT
The order of XJTU-China in IDT