| CPU_Nanjing - iGEM 2022

| CPU_Nanjing - iGEM 2022

Overview

Week 1 (December 6 to December 12, 2021) and
Week 2 (December 13 to December 19, 2021)

1. Recruiting members
2. Introducing to the laboratory of Prof. Hai Qian and meeting Dr. Xin Wang
3. Introducing laboratory safety issues
4. Making plans and preparing materials for interviews

Week 3 (December 20 to December 26, 2021) and
Week 4 (December 27, 2021 to January 2, 2022)

1. Education: preparing online public lessons

Week 5 (March 15 to March 21, 2022 (the same below))

1. Recruiting more members and building team CPU_Nanjing
2. Learning and examining the information about lab safety
3.Instructing to all team members about iGEM competition in detail

Week 6 (March 22 to March 28)

1. Brainstorming on the theme of our project
2. Analyzing and discussing feasibility by literature research
3. Integrated Human Practice (IHP): interviewing Dr. Xingxia Chen online

Week 7 (March 22 to March 28)

1. Designing a questionnaire based on suggestions form Dr. Chen
2. IHP: delivering online and offline questionnaire surveys

Week 8 (March 29 to April 4)

1. Analyzing statistics from our questionnaire survey
2. Human Practice (HP): interviewing Dr. Weimin Zhang
HP: bringing back phosphate-free soil from Changxing county

Week 9 (April 5 to April 11)

1. HP: interviewing Dr. Jierong Jiang
2. Discussing with team members about more actions related to HP
3. Determining the theme of our project based on HP/IHP

Week 10 (April 12 to April 18)

1. Purchasing chemicals for the experiments of our project
2. Preparing apparatus, including purchasing new equipment and transporting equipment from other labs (possessed by our PIs or instructors)
3. Synthesizing phosphite dehydrogenase gene in our lab

Week 11 (April 19 to April 25)

1. Cloning molecules and constructing bacterial strain
2. Holding a plenary meeting to plan the timeline of our project

Week 12 (April 26 to May 2)

1. Testing phosphite oxidation by engineered bacterial strains preliminarily
2. Discussing about experimental results and the "inherent laziness" of bacteria
3. IHP: interviewing Dr. Xin Wang again for further suggestions

Week 13 (May 3 to May 9)

1.Organizing the next part of our lab and designing our strategy based on suggestions provided by Dr. Xin Wang from our interview
2. Operating the site-directed mutagenesis of polyphosphate kinase
3. Creating our first Part from polyphosphate kinase

Week 14 (May 10 to May 16)

1. Collaboration: holding a meeting with team NJTech_China
2. Discussing our strategy about equilibrium expression
3. Collecting medium-copy number pBBR1MCS2 produced by our lab

Week 15 (May 17 to May 23)

1. Designing a vector co-express polyphosphate kinase and phosphite dehydrogenase
2. Constructing our vector and strain

Week 16 (May 24 to May 30)

1.Testing the enhancement of phosphite oxidation
2. Witnessing the intermediate polyphosphate for the first time
3. Proof of concept (POC): developing quick polyphosphate staining method

Week 17 (May 31 to June 6)

1. Collaboration: team NNU-China introducing us to their lab
2. Discussing the order of gene configuration on our vector
3. POC: testing the staining method by both labs led by Prof. Liu and Prof. Yang

Week 18 (June 7 to June 13)

1. Engineering: order of polyphosphate kinase and phosphite dehydrogenase on vector
2. Education: holding a plenary meeting to plan for more actions related to Education

Week 19 (June 14 to June 20)

1. Collaboration: holding an online meeting with ShanghaiTech_China
2. Discussing the possible challenges for project landing on terrestrial planets
3. Education: designing and producing materials used in our Education

Week 20 (June 21 to June 27)

1. IHP: interviewing Dr. Xiaomeng Wang
2. Drafting and revising our Preliminary Safety Form

Week 21 (June 28 to July 4)

1. Partnership: meeting with NJTech_China offline, determining our common goal
2. Education: entering high school classrooms, Qingjiang Middle School in Huaian

Week 22 (July 5 to July 11)

1. Engineering: discussing Engineering Success based on the comparison test completed in our lab previously
2. Witnessing enhanced polyphosphate accumulation in bacterial cells for the first time
3. POC: discussing the possibility that polyphosphate serves as flame retardants

Week 23 (July 12 to July 18)

1. Partnership: brainstorming online to seek for research subject
2. Discussing the biosafety of blue-green algae used in our project
3. Purchasing blue-green algae, non-toxic Microcystis aeruginosa FACHB-469

Week 24 (July 19 to July 25)

1. Finishing a systematic experiment on phosphate production using SMW medium and phosphite
2. Obtaining the final product in the form of high-concentration solution for the first time
3. POC: producing polyphosphate under open (unsterile) conditions
3. Creating the second and the third new Part

Week 25 (July 26 to August 1)

1. Cultivating algae
2. Verifying the general applicability of our design strategy by creating the fourth new Part
3. Finishing a systematic experiment performed on Parts to show our improvement

Week 26 (August 2 to August 8)

1. Partnership: making research on dealing with algae separately
2. IHP: cooking the algae (inspired by Dr. Xin Wang)
3. Testing the utility of algae lysate

Week 27 (August 9 to August 15)

1. Hardware: holding a plenary meeting to plan the Hardware design for bench-scale phosphate production
2. IHP: interviewing manager Zheng, Nanjing Organic Glass Factory
3. Designing solar-energy-driven sequence batch bioreactor

Week 28 (August 16 to August 22)

1. Education: visiting Purple Mountain Observatory with high school students
2. Finishing a systematic experiment on phosphate production using algae lysate
3. Education: communicating with public using WeChat official account and Bilibili account

Week 29 (August 23 to August 29)

1. Preparing the promotion video of our project
2. Partnership: confirming the effectiveness of algae lysate with NJTech_China
3. POC: preparing a series of media in different forms using algae as the raw material

Week 30 (August 30 to September 5)

1. Collaboration: bringing engineered strains to NNU-China for test
2. POC: testing the utility on clarified algae lysate by labs of Prof. Liu and Prof. Yang

Week 31 (September 6 to September 12)

1. Education: stepping into college campus with a new communication form of "ask and answer"
2. Hardware: debugging our equipment and starting bench-scale phosphate production
3. Implementation: cultivating lettuce by using the produced phosphate

Week 32 (September 13 to September 19)

1. Education: communicating with people in communities, introducing our whole project
2. Holding a plenary meeting to make a complete conclusion to our project
3. Modeling: starting the modeling process based on experiment data obtained in our lab

Week 33 (September 20 to September 26)

1. Partnership: dining together with NJTech_China, celebrating the success of our partnership and the completion of each other's project
2. Holding a plenary meeting to plan the design of our wiki, including artistic style, main color, font size and so on

Week 34 (September 27 to October 3)

1. Concluding all results obtained by different groups (experiment, human practices, presentation and modeling)
2. Bringing our Parts to complete the registration

Week 35 (October 4 to October 12)

1. Completing the construction of our wiki
2. Discussing the plan for our presentation video