Difference between revisions of "Team:Nanjing-China/Notebook"

Our team was founded this week! We met and communicated with each other. Through our team leader’s presentation, we knew IGEM a lot. Each member defined his/her mission in the team. We broadly read literature and brainstormed this year’s project. At first we came up with several different ideas, but later we reached an agreement that the most interesting and meaningful one was about nitrogen fixation.

Human Practices,Collaboration&Society:

We learned about nitrogenous fertilizer production status in quo and the advancement of China’s nitrogenous fertilizer industry. In order to better understand the actual demand of nitrogenous fertilizer in agriculture, we decided to visit farmers in Xiaohe Bei Village.
Having learned of the dearth of efficient and affordable fertilizer, we spared no effort to seek a cost effective nitrogen fixation method. Inspired by our previous work(Nanjing-China 2016), we creatively proposed an idea of “whole-cell photocatalytic nitrogen fixation”.
We helped Nanjing Forestry University to build their team.
We held conferences with Nanjing Agricultural University and China Pharmaceutical University to share experiences of being IGEMers.

Technical works Wet&Dry labs:

Wet Lab:Having confirmed the theme of our project, we began to work on our design. We read latest papers about biological nitrogen fixation, focused on the method sections and discussed what we didn’t understand in details. During the last week of this month, we worked out the first version of our design.
Dry lab:We communicated and exchanged ideas frequently in order to identify possible modeling directions which could provide useful guidance to our wet experiments. Later we proposed a few directions. The idea of developing homologous modeling of nitrogenase didn’t work successfully because we couldn’t get access to relevant software.

Human Practices,Collaboration&Society:

We planned to investigate the current production of nitrogenous fertilizer so we prepared interview questions and contacted Yantai Wuzhou Feng Fertilizer Plant. Then we went there, met the manager and were shown around the factories. We communicated with the technical R&D personnel and realized the big challenge we had to overcome before putting our project into practical application.

Technical works Wet&Dry labs:

Wet lab:We measured the transcriptional activity of nif promoter and constructed the complete line of nif cluster, BBa_K1796015. Then we transformed the plasmid Pcb1C3 containing the nif cluster and the fusion protein expression plasmid including E. coli outer membrane protein OmpA and the PbrR protein into E. coli strain pUC57. Besides,based on our Human Practice, we modified our design by adding Cd2+ toxicity test to it.

Exam Break

Human Practices,Collaboration&Society:

We invited Professor Haoqian Zhang and held a meet up with iGEM teams in Nanjing.
We were interviewed by Nanjing University Student Career Guidance Center. The Wechat Push introducing our team was issued on the public account “NJU Employment”.

>Technical works Wet&Dry labs:

Wet lab:We finished the last step in construction of whole-cell photocatalytic nitrogen fixation system----biosynthesis of CdS semiconductor. Then we performed gas chromatography to detect the amount of acetylene reduced to indirectly test the nitrogen fixation activity of our system.
Meanwhile,we conducted Real-time Quantitative PCR(QPCR).
Dry lab:Enlightened by the different relative transcriptional level of each nitrogenase component which was shown in the result of QPCR, we turned our attention to the extreme complexity of nitrogenase system. We perused literature on the stoichiometry of nitrogenase components.

Human Practices,Collaboration&Society:

We distributed brochures about our project at NJU.
We attended the 5th Conference of China iGEMer Community at ShanghaiTech University to demonstrate our project to all teams in China and learn from others.
We helped Central South University to found team.

Technical works Wet&Dry labs:

Wet lab:We conducted ICP-MS measurement of Cd2+ adsorption, Cd2+ toxicity test and TEM-EDX analysis. In addition,we improved our part to make it easier to operate.
Dry lab:We finally decided to model on the best stoichiometry of nif gene cluster. We looked through many common algorithm and figured out two modeling methods. After further comparation, we finally chose a method similar to greedy algorithm. We drew a flow diagram to describe the core idea of our method and as a reference for programming. Then we programmed with python, debugged our code and received the result.

Human Practices,Collaboration&Society:

We performed Language project with IIT Madras and received the finished video a few days later.
We helped another IGEM team, CSU-China to establish their team. We issued our Emoji chanllenge on the official website and received some interesting feedback shortly after that.

Technical works Wet&Dry labs:

Hardware: Inspired by our Human Practice, we formed an idea of designing a device for the growth of engineered E.coli strain. First we drew a draft on paper and then used the software solidworks to draw a 3D version draft. Eventually, a real device came out. The device provided a great help to our further experiments.

Dry lab:We refined our model.