Team:SZPT-CHINA/Notebook

● week 4(1.17-1.23)
Project:
1. Built a team.
2. Meeting to communicate the future work direction of our team.
● week 5(1.24-1.30)
Project:
1. The principal investigators taught the basic knowledge of synthetic biology for the team members, and set up courses on the learning app to upload relevant materials for everyone to learn at any time.
● week 6(1.31-2.6)
Project:
1. The principal investigators continued to teach the members the basic of synthetic biology.
2. The advisers guided us to learn about art skills, such as graphic design, video edit.
3. Brainstormed and collected everyone's ideas.
● week 7(2.7-2.13)
Project:
1. The team members browsed and studied a lot of outstanding projects in previous years during the winter vacation.
2. The principal investigators helped the team members learn the knowledge of synthetic biology through online courses.
3. Have the first online meeting to brainstorm, exchange ideas and clash of viewpoints.
● week 8(2.14-2.20)
Project:
1. We witnessed a marked growth of fire accidents in the news and paid attention to the problems with burn treatment.
2. We extensively consulted the literature to search for global burn data and clinical burn treatment methods.
● week 9(2.21-2.27)
Project:
1. Having the second online meeting to analyze the results of the brainstorming, screen out feasible ideas, and conduct preliminary research on them.
● week 10(2.28-3.6)
Project:
1. Reported the results of skills learning and presented the analysis of previous projects in the form of groups.
2. We continued to learn the knowledge of synthetic biology.
● week 11(3.7-3.13)
Project:
1. The team was divided into three groups: experimental group, human practice group and technical group.
Lab:
1. The experimental group learned the laboratory safety regulations and began experimental skill training.
● week 12(3.14-3.20)
Project:
1. Learned genetic engineering design under the guidance of instructor.
2. Participated in Ambassador Calls: iGEM 2021 Asian Community Meeting.
Lab:
1. The experimental group continued to learn experimental skills.
● week 13(3.21-3.27)
Project:
1. Each groups planed future work.
● week 14(3.28-4.3)
Project:
1. Conducted literature review and data research.
● week 15(4.4-4.10)
Project:
1. Conducted literature review with regard to Pseudomonas aeruginosa.
Human practice:
1. Interviewed firefighters to learn the cause of the fire.
2. Interviewed with the burn doctor of Shenzhen Longgang Central Hospital to consult about burn treatment methods, clinical drugs and other issues. And learned that the most common causative agente of burn infection is bacterial infection, among which Pseudomonas aeruginosa has a strong intrinsic drug resistance and the ability to produce drug resistance.
● week 16(4.11-4.17)
Project:
1. Confirmation that the anti-infective target bacteria group is Pseudomonas aeruginosa.
Human practice:
1. We consulted with Dr. Jun Li, Department of Burn and Skin Surgery of Xijing Hospital in an interview online.
● week 17(4.18-4.24)
Project:
1. It was found in the literature that Pseudomonas aeruginosa infection may also occur in chronic wounds.
● week 18(4.25-5.1)
Human practice:
1. Offline interviewed with Dr. Taofang He, Department of Chronic Wounds of the Affiliated Hospital of Shenzhen Southern University of Science and Technology to learn about the infection of Pseudomonas aeruginosa in chronic wounds.
2. We consulted with Dr. Jinyu Zhu, Department of Orthopedics and Joint Surgery, Affiliated Hospital of Southern University of Science and Technology in an interview online.
● week 19(5.2-5.8)
Project:
1. After careful consideration, we decided to carry out the project in the treatment of burn wounds.
Human practice:
1. In collaboration with associations in Shenzhen Polytechnic, we held the first of the fluorescent protein DIY activity in a school laboratory, introducing synthetic biology.
● week 20(5.9-5.15)
Human practice:
1. Held the second and third of the fluorescent protein DIY activity.
2.Popularized burn knowledge in Buji community, Longgang District, Shenzhen, Guangdong Province.
● week 21(5.16-5.22)
Human practice:
1. Offline interviewed with Dr. Dongyuan Sun, Department of Burn of Shekou People's Hospital in Shenzhen.
2. We consulted with doctors who from the Department of Burn in northern China in an interview online.
● week 22(5.23-5.29)
Project:
1. According to relevant literature, we started to design the module of Antipseudomonal Drug Synthesis under the guidance of our instructor.
Human practice:
1. Conducted a lecture on synthetic biology in Shenzhen Guangming District High School.
● week 24(6.6-6.12)
Project:
1. We decided to use bacterial cellulose as the material of wound dressings. This is because biological dressings can reduce infection, improve wound healing quality, alleviate patient pain and suffering compared with traditional wound dressings.
2. It was found in the literature that visible light is widely used in the medical field, so we were considering using the new-fangled methods of optogenetics as a gene circuit design method. we started to design the module of C-di-GMP Signaling and BC Film Production under the guidance of our instructor.
Human practice:
1. Communicated with Dr. Mingxing Tang in a school lab.
● week 25(6.13-6.19)
Project:
1. Participated in the round-table meeting held by CCIC.
2. Production of BC film was performed using Gluconacetobacter hansenii ATCC 53582 and make it as the host chassis.
3. We had finished all the design of the module of Antipseudomonal Drug Synthesis.
Human practice:
1. Paid a return visit to Dr. Weijia Ding from the Burns Department of Longgang Central Hospital in Shenzhen, asking the doctor about the feasibility of bacterial cellulose and the use of existing wound dressings.
2. Communicated with the Professor Jin Fan, he introduced applications of optogenetic to us.
● week 26(6.20-6.26)
Project:
1. Worked with our instructor to improve the gene circuit design of the module of c-di-GMP signaling and BC film production.
Human practice:
1. Opened a new section of synthetic biology on the WeChat official account.
Lab:
1. Planed experimental details and started experiments of the module of C-di-GMP Signaling and BC Film Production.
2. Constructed a series of parts for c-di-GMP synthesis enzyme and hydrolysis enzyme in E. coli DH5α.
● week 27(6.27-7.3)
Project:
1. Here we engineered Gluconacetobacter hansenii ATCC 53582 with a light-activated transcriptional promoter pDawn to optically regulate expressions of a lysis protein, taking into account of the actingmeachanism of antipseudomonal drugs and environmental and safety issues.
2. After finishing all the gene circuit design of 3 modules, we determined to name them after Antipseudomonal Drug Synthesis module, C-di-GMP Signaling and BC Film Production module, Safety and Drug Release module.
Human practice:
1. Made a synthetic biology summer camp plan for students on campus.
Lab:
1. A series parts of c-di-GMP synthesis enzyme and c-di-GMP hydrolysis enzyme was transferred into Gluconacetobacter hansenii ATCC 53582 with electrotransformation method.
2. PCR products were analyzed using 1% agarose gel electrophoresis to identify bands of expected sizes.
3. Minimum inhibitory concentration (MIC) was determined to use different antibiotics in Gluconacetobacter hansenii ATCC 53582.
● week 28(7.4-7.10)
Project:
1. We were designing the main wiki page.
Lab:
1. We did an experiment to screen out the best medium for Gluconacetobacter hansenii ATCC 53582 growth.
2. We did an experiment to explore the BC film production of Gluconacetobacter hansenii ATCC 53582 in SOC and HS medium, using J23100-mcherry-pSEVA331-G.hansenii ATCC 53582 and Gluconacetobacter hansenii ATCC 53582.
● week 29(7.11-7.17)
Project:
1. Completed the promotion video.
Human practice:
1. The members of the team carried out the project of bringing home synthetic biology and fire prevention knowledge, to preach our project and knowledge of synthetic biology in urban villages in Guangzhou, Xiamen, and Chaoshan respectively.
2. More than 70 college students were convened to give lectures on synthetic biology in the classroom offline.
3. Started a one-week synthetic biology summer camp, which would provide basic knowledge and laboratory skills in popular science microbiology and genetic engineering for college students.
Lab:
1. In order to improve the efficiency of electrotransform plasmid into Gluconacetobacter hansenii ATCC 53582, we did an electrotransformation condition exploratory experiment. Finally, 3kV was selected as the condition of our following experiments.
2. Screen the parts of c-di-GMP synthesis enzyme and c-di-GMP hydrolysis enzyme through exploring the BC film production of Gluconacetobacter hansenii ATCC 53582 in 12-well plates.
● week 30(7.17-7.24)
Human practice:
1. Connected with Professor Haifeng Ye from East China Normal University, and asked about the optimization of near-infrared light parameters.
2. Communicated with Xili Fire Squadron and discovered their learning needs of hazardous chemicals.
3. We conducted three courses at Yang Qi Culture and Arts Training Institution.
Lab:
1. Here we engineered E. coli DH5α with a light-activated transcriptional promoter pDawn to optically regulate expressions of S105(BBa_K3740024), φX174 E (BBa_K2656015) and Lysis Cassette LKD16 (BBa_K3740032), which uses a random primer labeling method.
2. The lysis strains were preliminarily screened out with 470nm blue light.
● week 31(7.25-7.31)
Project:
1. We chose BphS as the c-di-GMP synthetase, FcsR as the c-di-GMP hydrolase in our system.
2. The light-activated transcriptional promoter pDawn could response to blue light and induce expression in Gluconacetobacter hansenii ATCC 53582.
Human practice:
1. We invited a professional with experienced in information dissemination and social organizations to guide us in communication and public outreach of synthetic biology.
2. Participated in the second round-table meeting held by CCIC.
3. Listed chemicals that should not be watered in case of fire for firefighters on the WeChat official account.
4. We invited firefighters to conduct fire drills in the community's plaza, while our team members promote knowledge of burns.
Lab:
1. Verified the parts of c-di-GMP synthesis enzyme and c-di-GMP hydrolysis enzyme through exploring the BC film production of Gluconacetobacter hansenii ATCC 53582 in 12-well plate repeatedly.
2. The vector with the target lysis gene was transferred into G. hansenii ATCC 53582 with electrotransformation method.
3. Verified the function of pDawn promoter in G.hansenii ATCC 53582 in response to blue light.
● week 32(8.1-8.7)
Project:
1. Participated in the Southern China Regional Meeting.
2. Considered that the product is realized in the form of live bacteria.
3. The lysis protein that proves the lysis effect in Gluconacetobacter hansenii ATCC 53582 is X174 E and Lysis Cassette LKD16.
Human practice:
1. It has been found in the literature that Bacillus subtilis spray is suitable for the prevention and treatment of medium and small area second-degree burns and scald wound infections.
2. According to the game mode of 2048MINI game, we made the "Synthetic BC Film" mini game.
Lab:
1. We constructed a series of light-controlled complex parts in E. coli DH5α, using the c-di-GMP synthetase BphS and hydrolase FcsR.
2. Verified the ability of different lysis proteins to play a role in Gluconacetobacter hansenii ATCC 53582.
● week 33(8.8-8.14)
Project:
1. The prototype of the planned product is the live bacteria wound dressings.
2. It was proved that the protein that can stably perform the function of lysis in Gluconacetobacter hansenii ATCC 53582 is X174 E.
Human practice:
1. We visited Shenzhen Lando Biomaterials Co.,Ltd, to understand the application and production process of artificial skin.
2. We went to Dawang Xintian Village, an urban village in the Luohu District of Shenzhen, Guangdong Province, where we taught the children a picture book on synthetic biology of germ warfare and had English lessons. We also played a synthetic biology poker game and made drawings on paper plates together.
Lab:
1. The target gene expression vector of c-di-GMP light-controlled complex parts was transferred into G.hansenii ATCC 53582 with electrotransformation method.
2. PCR products were analyzed using 1% agarose gel electrophoresis to identify bands of expected sizes.
3. Verified the lysis function of X174 E and Lysis Cassette LKD16, which can play a role in Gluconacetobacter hansenii ATCC 53582 repeatedly.
● week 34(8.15-8.21)
Project:
1. There was a significant difference of the yield of BC under the condition of NIR light and dark for J23100-bphS-J23109-fcsR-pSEVA331-G.hansenii ATCC 53582 and J23100-bphS-J23110-fcsR-pSEVA331-G.hansenii ATCC 53582, but we can’t make G.hansenii ATCC 53582 completely do no BC film production under the dark condition.
Human practice:
1. We visited pharmacys near the school and ask about the existing burn treatment products and the forms of living bacteria products.
2. In order to determine the product format, we consulted Dr. Weijia Ding from the burn department about his views on live bacterial dressings once again.
3. We told stories for children in urban villages, using cartoon paper-cuttings.
Lab:
1. Verified a series of c-di-GMP light-controlled complex parts in Gluconacetobacter hansenii ATCC 53582 through exploring the BC film production in 12-well plate.
● week 35(8.22-8.28)
Project:
1. Determined the product form as a live bacteria by-product and preliminarily determine it be made into the emulsion.
Human practice:
1. We consulted with Xi'an Donghai Viable Mushroom Brand Promotion Co., Ltd, in an interview online.
2. Offline interviewed with Aegeli Eco-Technology Co., Ltd.
3. Issued a questionnaire to survey some questions related to living bacteria products.
4. We went to the urban villages, teaching children to fold DNA models and playing jigsaw puzzles of synthetic biology.
Lab:
1. Planed experimental details and started experiments on the Antipseudomonal Drug Synthesis module, then constructed a series of parts in E. coli DH5α.
2. Two plasmids S2-PET28A and SE-PET28A that were transferred into E. coli BL21 with electrotransformation method.
● week 36(8.29-9.4)
Project:
1. The wiki page design was finished.
Human practice:
1. We launched a charity sale activity with the theme of "The love of 'Kissed by light'".
2. Interviewed with Dr. Yan Xiong, a biological security expert, assessed whether the bacteria we used are dangerous.
Lab:
1. Verified the expression of S2 and SE protein by SDS-PAGE.
● week 37(9.5-9.11)
Project:
1. Proved that the SE-fusion protein can target Pseudomonas aeruginosa.
Human practice:
1. We launched a live broadcast for non-biological college students on the Bilibili platform. We mainly introduced the project and talked about synthetic biology relevant to our lives.
Lab:
1. Verified the SE-fusion protein whether could target Pseudomonas aeruginosa.
● week 38(9.12-9.18)
Project:
1. Determined to use pR-RBS300-SE-B1006-J23118-RBSII-IMM-rrB T1 (BBa_K3740050) as the final gene circuit of Antipseudomonal Drug Synthesis module.
Lab:
1. Carry out batch screening and verification experiments of SE-fusion protein for antipseudomonal performance.
● week39 (9.19-9.25)
Project:
1. Carried out data sorting work.
Human practice:
1. Organized product experience activities.
● week 40(9.26-10.2)
Project:
1. Started writing the wiki page.
● week 41(10.3-10.9)
Human practice:
1. Organized the fluorescent protein DIY activity again.
● week 42(10.10-10.16)
Project:
1. Wiki writing finished.