This year, ours program focuses on the abuse of antibiotics in society. Considering the reality of the situation and brainstorming in the team, we decided to solve the problem of antibiotics abuse by finding a substitute.
To begin with, we were inspired by TV reports about the dangers of antibiotic abuse. Further, we had access to a more comprehensive understanding of the overuse of antibiotics in society by attending the lecture, filed trips, releasing the questionnaire, and visiting relevant departments and companies.
Then, to figure out this problem, we consulted with some professors. At the same time, we also sought out other advisors to remove obstacle in our project, which is helpful for us to implement and refine our proposal.
Next, to make the project more accessible to the society, we organized offline public education activities on the topic of "synthetic biology", and distributed online questionnaires related to antibiotic abuse and our project.
In the process of all the above activities, we have ensured the information security of the participants according to the guidance of professionals.
In the whole HP activity, every step complements each other and forms an organic whole. You can click here to see more details.
Improvement of an Existing Part
We improve the part BBa_K3578010 from the 2020 iGEM worldshaper-Nanjing Team. Specifically, we characterized the 20 endogenous promoters in Y. lipolytica, and used the identified strong promoter P2 and P17 to express glucoamylase, which improved the ability of Y. lipolytica to utilize the starch. Moreover, we supplemented the experiments of shaking flask culture to confirm the new system's function. You can click here to see more details.
Our project aims to construct an enzyme-constrained model of Escherichia coli (BL21) to guide the production of antimicrobial peptides. Firstly, to verify the accuracy of enzyme-constrained model ec_iECBD_1354, we calculated the ability of using 14 kinds of carbon sources. After the verification of model ec_iECBD_1354, we further used it to identify the targets, which can improve the synthesis of SMAP protein. When comparing the differences of protein demand between cell growth stage and product synthesis stage, 7 proteins were identified as top-demand, which need to be up-regulated. However, 21proteins were classified as defined as reduced demand, which need to be down-regulated.
Finally, to test the results of model analysis, we selected the proteins AsnA and PurA. In detail, the asnA gene, encoding the protein AsnA, was overexpressed with the strong promoter of Pj23119 at the IS site in the genome of BL21 (DE3), yielding the engineered strain of asnA. And, the purA gene, encoding the protein PurA, was knocked out from the genome of BL21(DE3), yielding the engineered strain of △purA. And, we tested the growth of two engineered strains when no AMP was expressed. Results showed that two engineered strains all keep the similar growth with the wild strain.
Through the design of the final loop, we demonstrated some of the results we hoped to get when we originally designed the project, which you can find on our Proof of Concept page.
Because of the repeated COVID-19, our communication with Worldshaper-Nanjing took the way of combining online and offline. We hold meetings with Worldshaper-Nanjing at least once a month.
We all hope to carry out our projects in an innovative form. We have conducted in-depth cooperation in experimental design and implementation, HP, education, Wiki, etc. The cooperation between our two teams is of great help to the promotion of our projects. For more details about our parts, you can visit our Partnership page.
Education and Communication
We firmly believe that education and communication are a bridge of knowledge exchange between the public and researchers. This year, Nanjing Normal University has made many efforts to eliminate knowledge misunderstandings and attract public attention to synthetic biology and our programs . Due to the limited conditions of the epidemic, most of our science exchange activities were carried out online. Additionally, we also combine the offline scientific communication in different ways. In generally, we have increased public knowledge of IGEM, synthetic biology, and the life sciences.
As researchers, we consider it our responsibility to engage with society and show them first-hand information, either through the introduction of synthetic biology or our project. For more details on our Science Communication, you can visit our Science Communication page.
This year NNU-China 2021 has tackled and solved one or more of our project's problems and use synthetic biology tools to generate expected results.
After design improving through literature analysis and Human Practices (See the Human Practices details), our project constructed the BL21 (DE3)-derived variant strains library with different RBS sequences of T7RNAP, and got the best expression host for the desired antimicrobial peptides (AMPs) production by a high throughput host screening method. Moreover, as a proof-of-concept, one of the AMP, SMAP (RGLRRLGRKIAHGVKKYGPTVLRIIRIAG), was tested (See the Parts and Design details).
We have done a lot of amazing work to make sure that they can realize the goals we set. Moreover, it is worth mentioning that BBa_K3868097, and BBa_K3868100-109 are typical among them. These BioBricks work well to perform their functions and works as expected. All data related are recorded engineering success page. We hope they will make some contribution to the iGEM community. For more details about our parts, you can visit our Engineering Success page.
We spanned the limitations of space and time by holding online meetings, and achieved communication and help with other teams. You can click here to see more details.
Based on reasonable social investigation methodology, we conducted several human practice projects around our project and synthetic biology, sucha as literature search, interviews, and intercollegiate communications. You can click here to see more details.
This year, we are committed to developing high-quality antimicrobial peptides to replace antibiotics，which can solve the harmful problem of overuse of antibiotics and improve human health. We constructed an Escherichia coli library through synthetic biology methods o screen the optimal strain for antimicrobial peptides production. This method for screening the optimal strain for antimicrobial peptides production is convenient and quick. If the antimicrobial peptides produced by our engineered strains can be used in factories, hospitals, farms and other places, it can not only solve the problems caused by the abuse of antibiotics, but also ensure the quality of products and production scale.
We have or will meet all deliverables on the Competition Deliverables page.
We have created a separated page on our wiki showing the attribution of each aspect clearly. You can click here to see more details.
We have documented how we were inspired and finally selected our project on the “Project Description” page, and we also introduced how we originally achieved our goals. You can click here for more details.
We chose to characterize the previous BioBrick parts as our contribution in characterization. In detail, we completed the quantitative experimental characterization of BBa_K3645011, BBa_K346085, BBa_K3578000, and BBa_K2375000, and added the data of them to the corresponding Biobrick. You can click here for more details.