Attributions

What We Did In This Summer

In this summer, we aimed at degrading PET biologically with two kinds of newly-found enzymes PETase and MHETase.

We modified the enzyme PETase to improve its activity and used R-R (Reporting and Regulation) system to make the expression visible and regular.

We used microbial consortia to continue degrading the products TPA and EG to carbon dioxide.

We also introduced Cyanobacteria to the microbial consortia so that the system can work with only optical energy and carbon dioxide. We also use CFPS(cell-free protein synthesis) system to assay the enzyme activity.

In the Human Practice part, we spread out our project and the glamour of synthetic biology and receive financial help from some corporations.

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General Support

Our instructor Prof. Yingjin Yuan and Dr. Bingzhi Li: A huge thanks for their constructive suggestions and meticulous guidance to our project.

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Protein modification Support

Prof. Mingdong Yao, who helped us to analyze the stability of PETase, gave many advices and discussed with us the practicability of our principles of designing mutations of PETase.

Dr. Yi Wu, the man who generously shared restriction enzyme as well as watermelon, who gave us enlightening advices during showers, who got us a brand new microplate reader in the crisis, who helped novices to fix all kinds of experiment problems, and who supported the Tianjin iGEM team all the time.

Dr. ZeXiong Xie, who gave us advices and help on experiments all the time and supported Tianjin iGEM team.

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Bacteria Consortium Support

Prof. CunJiang Song generously sponsored us with P.putida KT2440 and the shuttle plasmid pBBR1MCS-2 which can be transformed into P.putida by electroporation.

Prof. Aijun Miao generously sponsored us with Rhodococcus sp. Strain RHA1.

Prof. Rui Ban generously sponsored us with Bacillus subtilis DB104.

Prof. Tao Chen generously sponsored us with Bacillus subtilis 168 and its available plasmids pH12-p43 and pH13-p43.

Prof. Jun Yang shared us with ideas about experimental design and measurement.

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Cyanobacteria Support

Prof. Weiwen Zhang who allowed us to use their laboratories and equipments, showed great patience when we occupied many of the facilities.

Dr. Jinjin Diao, at Tianjin University, helped us with his knowledge of Cyanobacterium Synechocystis sp. PCC 6803. He commented on our thoughts and ideas which helped us establishing the cell wall disruption process.

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CFPS(cell-free protein synthesis) System Support

Prof. Dayong Yang Professor at Tianjin University. He commented on our thoughts about using the cell-free system to express our enzymes and ideas. He also offered us the plasmid pRset_CFP-1 and allowed us to establish the expression system in his Lab.

Yang Liu postgraduate student in Prof. Dayong Yang’s Lab, who offered the material such as lysate of E.coli, feeding buffer and so on.

Yannan Zhao postgraduate student in Prof. Dayong Yang’s Lab, who assisted us to do some experiments and instucted us to use the ELIASA (microplate reader).

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Human Practice Support

GenScript sponsors us with 60000 CNY.

Synbio Technologies sponsors us with 15000 CNY.

AllTech sponsors us with 10000 CNY.

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