iGEM 2021 — Team Tsinghua

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# Team:Tsinghua/Notebook

# Notebook

Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.

### What should this page have?

* Chronological notes of what your team is doing.
* Brief descriptions of daily important events.
* Pictures of your progress.
* Mention who participated in what task.

### Inspiration

You can see what others teams have done to organize their notes:

* [2018 Munich](https://2018.igem.org/Team%3AMunich/Notebook)
* [2019 Georgia State](https://2019.igem.org/Team%3AGeorgia_State/Notebook)
* [2019 Newcastle](https://2019.igem.org/Team%3ANewcastle/Notebook)
* [2020 IISER Pune India](https://2020.igem.org/Team%3AIISER-Pune-India/Notebook)
* [2020 Lund](https://2020.igem.org/Team%3ALund/Notebook)
* [2020 NOVA LxPortugal](https://2020.igem.org/Team%3ANOVA_LxPortugal/Notebook)
* [2020 RDFZ China](https://2020.igem.org/Team%3ARDFZ-China/NoteBook)

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Tsinghua/Engineering

* [![Icon for Homepage](https://static.igem.org/mediawiki/2021/5/5c/T--Tsinghua--homepage_icon.png)](https://2021.igem.org/Team%3ATsinghua)
* [HOME](https://2021.igem.org/Team%3ATsinghua)
* [TEAM ▼](#team)
  + [Members](https://2021.igem.org/Team%3ATsinghua/Members)
  + [Attributions](https://2021.igem.org/Team%3ATsinghua/Attributions)
* [PROJECT ▼](#project)
  + [Description](https://2021.igem.org/Team%3ATsinghua/Description)
  + [Design](https://2021.igem.org/Team%3ATsinghua/Design)
  + [Pro-Lac](https://2021.igem.org/Team%3ATsinghua/Pro-lac)
  + [Mucosal Healing](https://2021.igem.org/Team%3ATsinghua/Mucosal_Healing)
  + [Microbiota Regulation](https://2021.igem.org/Team%3ATsinghua/Microbiota_Regulation)
  + [Metabolic Balancing](https://2021.igem.org/Team%3ATsinghua/Metabolic_Balancing)
  + [Delivery](https://2021.igem.org/Team%3ATsinghua/Delivery)
  + [Proof of Concept](https://2021.igem.org/Team%3ATsinghua/Proof_Of_Concept)
  + [Engineering](https://2021.igem.org/Team%3ATsinghua/Engineering)
* [SAFETY](https://2021.igem.org/Team%3ATsinghua/Safety)
* [EXPERIMENT ▼](#experiment)
  + [Parts](https://2021.igem.org/Team%3ATsinghua/Parts)
  + [Protocol](https://2021.igem.org/Team%3ATsinghua/Protocol)
  + [Lab Notebook](https://2021.igem.org/Team%3ATsinghua/Lab-notebook)
* [MODELING ▼](#modeling)
  + [Main Page](https://2021.igem.org/Team%3ATsinghua/Model)
  + [LAP Adhesion](https://2021.igem.org/Team%3ATsinghua/LAP_adhesion)
  + [Simulator](https://2021.igem.org/Team%3ATsinghua/Simulator)
  + [Lateral Flow](https://2021.igem.org/Team%3ATsinghua/Lateral_flow)
  + [Protein Degradation](https://2021.igem.org/Team%3ATsinghua/Protein_degradation)
* [SOFTWARE ▼](#software)
  + [P2N](https://2021.igem.org/Team%3ATsinghua/Software)
  + [gDesign](https://2021.igem.org/Team%3ATsinghua/gDesign)
* [HP ▼](#HP)
  + [Human Practices](https://2021.igem.org/Team%3ATsinghua/Human_Practices)
  + [Collaborations](https://2021.igem.org/Team%3ATsinghua/Collaborations)
  + [Partnership](https://2021.igem.org/Team%3ATsinghua/Partnership)
* [AWARD ▼](#award)
  + [Contribution](https://2021.igem.org/Team%3ATsinghua/Contribution)
  + [Medal Criteria](https://2021.igem.org/Team%3ATsinghua/award/medal-criteria)
  + [Education](https://2021.igem.org/Team%3ATsinghua/Education)
  + [Communication](https://2021.igem.org/Team%3ATsinghua/Communication)
  + [Implementation](https://2021.igem.org/Team%3ATsinghua/Implementation)

# Engineering

Engineering

* [Microcin Regulation Project](#Microcin)
* [Mucosal healing project](#Mucosal)
* [Metabolic balancing part](#Metabolic)

> We have constructed many parts successfully in microcin regulation project, mucosal healing project and in metabolic balancing part.
>
>
>
> # Microcin Regulation Project
>
> To construct a huge plasmid containing nine genes, we put them into four polycistrons: mcmK-mcmL, mchI-mchB-mchX, mchC-mchD, mchE-mchF. We had constructed them successfully.(Fig.1) For polycistrons containing the regulatory genes(mcmK-mcmL, mchC-mchD, mchE-mchF), we placed them under a constitutive expression promoter -- J23119. For mchI-mchB-mchX, we used Ptac-lacO to represent a regulatory promoter. Unfortunately, we failed to construct the final version plasmid, as the mcmKL failed to merge with other polycistrons.(Fig.1)
>
>
> ![](https://static.igem.org/mediawiki/2021/7/79/T--Tsinghua--Fig4._engineering_microcin_huge_plasmid.png)
>
> Fig. 1 The identification of four polycistrons in DNA level.
>
> # Mucosal healing project
>
> In order to heal the intestinal tract damaged by IBD, we adopted a special therapy expressing the therapeutic proteins by E.coli Nissle 1917 (EcN) in situ. The design is based on a ternary system: sensor - secretion peptide - therapeutic proteins. Thus, EcN can be induced by inflammatory signals to secrete therapeutic proteins in the damaged intestine, promoting the intestinal mucosal healing.
>
>
>
> For each of the three sections, we can replace with different parts freely for a more comprehensive treatment system. Therefore, there are various of combinations, making the whole method more flexible, so that we can provide the treatment with multi-aspects.
>
>
> ![](https://static.igem.org/mediawiki/2021/b/b6/T--Tsinghua--Fig3._engineering_construct.png)
>
> Fig.2 The design of sensor-secretion peptide-therapeutic proteins.
>
> CsgA and TFF1/2/3 were chosen separately as secretion peptide and therapeutic proteins. To ensure the existence of the plasmid in the nissle 1917, we perform clony PCR and agarose gel electrophoresis. The results are shown in Fig.2.
>
>
> ![](https://static.igem.org/mediawiki/2021/8/8b/T--Tsinghua--Fig4._engineering_tp.png)
>
> Fig.3 Identification of TFF1/2/3 in DNA level
>
> # Metabolic balancing part
>
> We had constructed BSH1 successfully. Fig.4 show the extraction of assembled BSH1 DNA. Lane 4,5,6,8 represented the plasmids containing N tagged BSH1 gene which was later sequenced.
>
>
> ![](https://static.igem.org/mediawiki/2021/0/06/T--Tsinghua--Fig5._engineering_BSH1.png)
>
> Fig4.The identification of BSH1 in DNA level.

Copyright © 2021 iGEM Team: Tsinghua. All rights reserved.

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Tsinghua/Experiments

# Experiments

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

If you made Parts this year, please remember to put all information, characterization, and measurement data on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).

### What should this page contain?

* Protocols
* Experiments

### Inspiration

* [2019 Nantes](https://2019.igem.org/Team%3ANantes/Experiments)
* [2019 TU Eindhoven](https://2019.igem.org/Team%3ATU_Eindhoven/Experiments)
* [2019 Mingdao](https://2019.igem.org/Team%3AMingdao/Demonstrate)
* [2020 Amsterdam](https://2020.igem.org/Team%3AAmsterdam/Experiments)
* [2020 NCTU Formosa](https://2020.igem.org/Team%3ANCTU_Formosa/Experiments)
* [2020 USAFA](https://2020.igem.org/Team%3AUSAFA/Experiments)

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Tsinghua/Results

# Results

You can describe the results of your project and your future plans here.

### What should this page contain?

* Clearly and objectively describe the results of your work.
* Future plans for the project.
* Considerations for replicating the experiments.

### Describe what your results mean

* Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for.
* Show data, but remember **all measurement and characterization data must also be on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).** Otherwise these data will not be in consideration for any medals or part awards!
* Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project.

### Project Achievements

You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.

* A list of linked bullet points of the successful results during your project
* A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.

### Inspiration

See how other teams presented their results.

* [2019 Newcastle](https://2019.igem.org/Team%3ANewcastle/Results)
* [2019 Munich](https://2019.igem.org/Team%3AMunich/Results)
* [2019 Tec Chihuahua](https://2019.igem.org/Team%3ATec-Chihuahua/Results)
* [2020 Aalto Helsinki](https://2020.igem.org/Team%3AAalto-Helsinki/Results)
* [2020 GreatBay SCIE](https://2020.igem.org/Team%3AGreatBay_SCIE/Results)
* [2020 Queens Canada](https://2020.igem.org/Team%3AQueens_Canada/Results)