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<p><b>Plasmid Standardization:</b></p> | <p><b>Plasmid Standardization:</b></p> | ||
− | <p>Plasmid standardization was completed by <b>Zhiqing Guo</b> and <b>Zhuoqi Huang</b> by using 3A | + | <p>Plasmid standardization was completed by <b>Zhiqing Guo</b>, <b>Hangxi Fu</b> and <b>Zhuoqi Huang</b> by using 3A |
Assembly. | Assembly. | ||
</p> | </p> |
Latest revision as of 02:00, 18 October 2018
Our project was designed by Zhujun Xia and all circuits were designed by Zhujun Xia, YinQing Zeng and Mo Qiqin.
sifA was knocked out by Zhujun Xia, Zhuoqi Huang and Wenxin Hu also gave assistance.
Intracellular Environment-Dependent Circuits:
The intracellular environment-dependent circuits were designed by Mo Qiqin and Zhujun Xia.
The intracellular environment-dependent circuits were constructed by Mo Qiqin. Ruonan Tian also gave assistance.
Chemical Control System:
The ATc induction chemical control system was constructed by He Yang and Zhuoqi Huang also gave assistance.
The TetR-eGFP fusion protein expression system for TetR expression measurement was constructed by He Yang and Zhuoqi Huang also gave assistance.
Targeting Specificity and Surface-Displaying:
RGD-inserted OmpA circuit clone and construction of Salmonella SL1344_RS05255 (OmpA coding sequence) were completed by YinQing Zeng and Lingyu Zhong.
The circuit of surface-display system was constructed by Lingyu Zhong. Optimation of OmpA in E. Coli was completed by Zhujun Xia.
Plasmid Standardization:
Plasmid standardization was completed by Zhiqing Guo, Hangxi Fu and Zhuoqi Huang by using 3A Assembly.
Plasmid Submission:
Submitted plasmids (Pcs2-eGFP-GSDMD FL, Pcs2-eGFP-GSDMD-N275 and two GSDMD full length mutations) were constructed by Zhiqing Guo and Wenxin Hu.
The description of submitted plasmids by ATP assay and cell phenotype via transfection was designed and completed by Zhujun Xia.
Bacterial phenotype verifications were designed and completed by He Yang, including the OD measurement and CFU measurement of chemical control system.
Cell phenotype verifications were designed and completed by Zhujun Xia, including the verification of chemical control system, and targeting specificity, all the related circuits and the demonstration of the final work system.
Safety verification was designed and completed by Zhujun Xia, which demonstrated our project is safe enough to be used.
InterLab was conducted by Xichen Rao, Zhujun Xia, Ruonan Tian and Heng Heng also gave assistance. Our data passed the check of the Measurement Committee successfully.
Mathematical model and app about salmonella infection and GSDMD release were built by Songtao Chen.
ATc model which predicted the relationship between concentration of ATc and concentration of GSDMD was built by Xichen Rao.
The modeling on adjusting the strength of RBS and ATc with fixed GSDMD-N concentration was completed by Yini Miao.
The modeling of the promoter sifA and promoter ent was completed by Ruonan Tian.
The applet of "Bacterial Colony Counter" was designed and programmed by Shuguang Wang.
Collaboration and communication with other teams were completed by Yinqing Zeng and all the other team members.
Handicraft manufacture was designed and completed by Yinqing Zeng.
Questionnaire preparation, the advertisement of synthetic biology and preparation of popular science articles were completed by Heng Heng.
Two posters for Eurasian exchange meetings and CCIC were completed by Ruonan Tian.
Our wiki framework was designed and programmed by Shuguang Wang and our artist were Yuying Xiang and Kening Chen.
Our blog was built by Shuguang Wang.
The plate reader machine operation was conducted by Mo Qiqin.
Chief cleaner of the laboratory was Xichen Rao.
For each parts of experiments, our appreciations are as follow:
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for sharing the cell lines of HEK293T, HELA, MCF7, iBMDM, so that our experiment can process successfully.
Thanks to Dr. Feng Shao’s laboratory, for sharing the cell line of GSDMD-/- HELA cells, which provides us great support.
Thanks to Dr. Yi Zeng and Dr. Ms Hong Fan, Shanghai Institute of Biochemistry and Cell Biology, CAS, for sharing the cell line of MDA-MB-231.
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for sharing the strain of Salmonella enterica var. Typhimurium SL1344.
Thanks to Dr. Chenli Liu from SIAT CSynBER for generously sharing the material (Strain: E.coli K-12 MG1655 with mCherry in the genome).
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for providing the protocol and material of gene editing based on two-step allelic exchange, which provides us with great help.
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for sharing the plasmid backbone (Pcs2-eGFP) to us.
Thanks to GeneScript and IDT for DNA sequence synthesis service.
Thanks to SnapGene and MathWorks for free software supporting.
![](https://static.igem.org/mediawiki/2018/c/cd/T--HZAU-China--GenScript.png)
![](https://static.igem.org/mediawiki/2018/6/6a/T--HZAU-China--IDT.png)
![](https://static.igem.org/mediawiki/2018/0/0e/T--HZAU-China--SnapGene.png)
![](https://static.igem.org/mediawiki/2018/5/5d/T--HZAU-China--MathWorks.png)
Thanks to College of Life Science and Technology and Huazhong Agricultural University for providing the working site and fund for us.
![](https://static.igem.org/mediawiki/2018/1/1e/T--HZAU-China--huanong.png)
Thanks to Prof. Binguang Ma as our Primary PI and for supervising the whole project and instructing our modeling.
Thanks to Prof. Jin He as our Secondary PI and Prof. Shan Li for instructing our experiment.
Thanks to Prof. Jin He, Prof. Shan Li, Prof. Gang Cao, Prof. Donghai Peng and Prof. Ming Sun for giving us advices at the proposal presentation.
Thanks to Kening Chen, Wenqi Huang and Haimeng Li from iGEM-2017, Huazhong Agricultural University, for giving us advices.
Thanks to Weitong Guo from iGEM-2017, Huazhong Agricultural University, for the training of new dry-labers during winter vacation.
We really appreciate your support!