iGEM 2020 — Team Shanghai_HS_United

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Shanghai HS United/Notebook

非模块化方式使用layui

Entprenuership

![](https://static.igem.org/mediawiki/2020/c/c6/T--Shanghai_HS_United--homepage03.png)

**GalAcid**

---

* [Home](https://2020.igem.org/Team%3AShanghai_HS_United)
* Project

  :   [Description](https://2020.igem.org/Team%3AShanghai_HS_United/Description)
  :   [Design](https://2020.igem.org/Team%3AShanghai_HS_United/Design)
  :   [Experiments](https://2020.igem.org/Team%3AShanghai_HS_United/Experiment)
  :   [Notebook](https://2020.igem.org/Team%3AShanghai_HS_United/Notebook)
  :   [Result](https://2020.igem.org/Team%3AShanghai_HS_United/Result)
  :   [Safety](https://2020.igem.org/Team%3AShanghai_HS_United/Safety)
* [Contribution](https://2020.igem.org/Team%3AShanghai_HS_United/Contribution)
* [Engineering](https://2020.igem.org/Team%3AShanghai_HS_United/Engineering)
* [Implementation](https://2020.igem.org/Team%3AShanghai_HS_United/Implementation)
* Parts

  :   [Basic Parts](https://2020.igem.org/Team%3AShanghai_HS_United/Basic%20Parts)
  :   [Composite Parts](https://2020.igem.org/Team%3AShanghai_HS_United/Composite%20Parts)
  :   [Parts Collection](https://2020.igem.org/Team%3AShanghai_HS_United/Parts%20Collection)
* [Model](https://2020.igem.org/Team%3AShanghai_HS_United/Model)
* Human Practice

  :   [Integrated Human Practice](https://2020.igem.org/Team%3AShanghai_HS_United/Human_Practices)
  :   [Fundraising](https://2020.igem.org/Team%3AShanghai_HS_United/Fundraising)
  :   [Entrepreneurship](https://2020.igem.org/Team%3AShanghai_HS_United/Entrepreneurship)
* Team

  :   [Team Member](https://2020.igem.org/Team%3AShanghai_HS_United/Team)
  :   [Collaborations](https://2020.igem.org/Team%3AShanghai_HS_United/Collaborations)
  :   [Attributions](https://2020.igem.org/Team%3AShanghai_HS_United/Attributions)

Notebook

20200808

Professor
Yang gave us an online presentation about the project.

20200812

All
the students gathered in Shanghai, had ice-breaking activities, designed our logo, did self-introduction and
selected team leader candidates.

![](https://static.igem.org/mediawiki/2020/6/68/T--Shanghai_HS_United--notebook01.jpg)

20200813

First
day in the lab, we learned safety precautions and got familiar with experimental equipment. Then did PCR to clone
genes and gel extraction.

20200815

We
constructed one of our plasmids and sequenced in a company.

20200817

We
constructed another plasmid and sequenced in a company.

20200818

The two plasmids are transformed into E. coli competent
cell.

20200819

We did
fermentation experiments in a conical flask.

we
also collaborated with RePhos team.

Expressed
protein.

Sugar
consumption and butyric acid formation were determined

20200820

Samples
were taken to determine sugar consumption and butyric acid production.

We
also took our team photos.

20200821

Samples
were taken at anther time-point to determine sugar consumption and butyric acid production.

20200822

Analyze
experimental data, summarize and answer questions.

20200823

We
did live presentation on zoom software then
finished our training.

Contact Email: 3142451726@qq.com

Wechat: Search "2020 IGEM CounterSugar" in WeChat

School Website: https://www.xdfyz.cn/

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Shanghai HS United/Engineering

非模块化方式使用layui

Entprenuership

![](https://static.igem.org/mediawiki/2020/c/c6/T--Shanghai_HS_United--homepage03.png)

**GalAcid**

---

* [Home](https://2020.igem.org/Team%3AShanghai_HS_United)
* Project

  :   [Description](https://2020.igem.org/Team%3AShanghai_HS_United/Description)
  :   [Design](https://2020.igem.org/Team%3AShanghai_HS_United/Design)
  :   [Experiments](https://2020.igem.org/Team%3AShanghai_HS_United/Experiment)
  :   [Notebook](https://2020.igem.org/Team%3AShanghai_HS_United/Notebook)
  :   [Result](https://2020.igem.org/Team%3AShanghai_HS_United/Result)
  :   [Safety](https://2020.igem.org/Team%3AShanghai_HS_United/Safety)
* [Contribution](https://2020.igem.org/Team%3AShanghai_HS_United/Contribution)
* [Engineering](https://2020.igem.org/Team%3AShanghai_HS_United/Engineering)
* [Implementation](https://2020.igem.org/Team%3AShanghai_HS_United/Implementation)
* Parts

  :   [Basic Parts](https://2020.igem.org/Team%3AShanghai_HS_United/Basic%20Parts)
  :   [Composite Parts](https://2020.igem.org/Team%3AShanghai_HS_United/Composite%20Parts)
  :   [Parts Collection](https://2020.igem.org/Team%3AShanghai_HS_United/Parts%20Collection)
* [Model](https://2020.igem.org/Team%3AShanghai_HS_United/Model)
* Human Practice

  :   [Integrated Human Practice](https://2020.igem.org/Team%3AShanghai_HS_United/Human_Practices)
  :   [Fundraising](https://2020.igem.org/Team%3AShanghai_HS_United/Fundraising)
  :   [Entrepreneurship](https://2020.igem.org/Team%3AShanghai_HS_United/Entrepreneurship)
* Team

  :   [Team Member](https://2020.igem.org/Team%3AShanghai_HS_United/Team)
  :   [Collaborations](https://2020.igem.org/Team%3AShanghai_HS_United/Collaborations)
  :   [Attributions](https://2020.igem.org/Team%3AShanghai_HS_United/Attributions)

Research -> Image -> Design

As a primary part of lactose, galactose is found in most galactose products and many baby formulas. But it is dangerous for galactosemia patients to eat galactose products. Galactosemia is an autosomal recessive disorder disease leading to abnormal in using galactose to produce energy. The patients have galactose 1-phosphate uridyltransferase (GALT) deficiency. There are three kinds of this disease: Type 1 galactosemia occurs in 1 out of every 6000-10000 babies, Type 2 galactosemia is less common than type 1 and occurs in 1 out of every 100,000 babies, Type 3 and Duarte variant galactosemia are very rare. Long-term outcome for older patients with galactosemia remains problematic although neonatal hepatotoxic syndrome is prevented by newborn screening and galactose restriction.

![](https://static.igem.org/mediawiki/2020/3/36/T--Shanghai_HS_United--engineering01.jpg)

Based on the information presented above, we came up with the plan using engineering probiotics to degrade and digest the galactose in galactosemia patients. Our goal is to construct an engineering E. Coli Nissle 1917, which carries a butyrate synthesis path gene cluster with J23100 promoter (Part: BBa\_K3585003). The engineering bacterial can transform galactose to butyrate, that is beneficial for galactosemia patients.

Build -> Test

When cultured with glucose as the carbon source, the glucose was almost run out till cultured for 8 hours. When the culture medium contained glucose and galactose, the carbon source consumption rate was lower than that of the culture medium containing only glucose. However, the bacteria had butyrate synthesis gene, the consumption rate of glucose was accelerated.

![](https://static.igem.org/mediawiki/2020/d/df/T--Shanghai_HS_United--engineering02.jpg)

Figure 1. the concentration of glucose when supplied with glucose. Note: each of the strains we have a repeat. Glu represents that we use glucose as the carbon source while fermentation; GG represents that we use both glucose and galactose as the carbon source while fermentation; 1917 represents wild type strains; pButy represents that the host strain contained pMTL83151-J23100-butyrate plasmids and expressed butyrate synthesis gene cluster; placY represents that the host strain contained p15A-J23200-lacY plasmids and expressed lacY gene; 1 and 2 represent for sample repeats.

Learn -> Research -> Improved design

According to the articles, the presence of glucose inhibits the synthesis of many catabolic enzymes, a phenomenon called "glucose effect" or "glucose inhibition". Studies have shown that overexpression of LacY can eliminate the glucose effect. We design to overexpress lacY protein to achieve simultaneous metabolism of glucose and galactose, thereby increasing the efficiency of degrading galactose.

The second round of Build -> Test

Bacteria with lacY was able to grow using galactose as the unique carbon source in the culture, although the growth rate of the host bacteria is obviously slower than those strains cultured with glucose.

![](https://static.igem.org/mediawiki/2020/3/30/T--Shanghai_HS_United--engineering03.jpg)

Figure 2. Growth rate of host strains when supplied with different carbon sources. Each of the strains we have a repeat. Glu represents that we use glucose as the carbon source while fermentation; Gal represents that we use galactose as the carbon source while fermentation; GG represents that we use both glucose and galactose as the carbon source while fermentation; 1917 represents wild type strains; pButy represents that the host strain contained pMTL83151-J23100-butyrate plasmids and expressed butyrate synthesis gene cluster; placY represents that the host strain contained p15A-J23200-lacY plasmids and expressed lacY gene; 1 and 2 represent for sample repeats.

When the medium contained only galactose, the host bacteria expressing lacY gene began to consume lactose at 4 hours, indicating that lacY gene could promote the consumption of galactose by the host bacteria.

![](https://static.igem.org/mediawiki/2020/b/bf/T--Shanghai_HS_United--engineering04.jpg)

Figure 3. Consumption rate of galactose. Each of the strains we have a repeat. Gal represents that we use galactose as the carbon source while fermentation; GG represents that we use both glucose and galactose as the carbon source while fermentation; 1917 represents wild type strains; pButy represents that the host strain contained pMTL83151-J23100-butyrate plasmids and expressed butyrate synthesis gene cluster; placY represents that the host strain contained p15A-J23200-lacY plasmids and expressed lacY gene; 1 and 2 represent for sample repeats.

The second round of Learn -> Research -> Improved design

Based on the experiment information presented above and more related research articles, we came up with the new improved design engineering part BBa\_K3585007, that has a butyric acid synthesis pathway along with a lac-Y gene that helps the probiotic to prioritize the ingestion of galactose.

We design a strong promoter J23200 to continuously overexpress lac-Y gene to achieve simultaneous metabolism of glucose and galactose. Continuous expression of butyrate synthesis pathway enzymes through a constitutive promoter, so the strain can degrade galactose and synthesize butyrate. Combining the butyrate synthesis pathway enzyme and lac-Y, the engineered strain can effectively degrade galactose in the presence of glucose.

Contact Email: Austin1001@qq.com

Wechat: Search "iGEM GalAcid团队" in WeChat

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Shanghai HS United/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

* [2014 Colombia](https://2014.igem.org/Team%3AColombia/Protocols)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Protocols)
* [2014 Caltech](https://2014.igem.org/Team%3ACaltech/Project/Experiments)

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:Shanghai HS United/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.

* [2014 TU Darmstadt](https://2014.igem.org/Team%3ATU_Darmstadt/Results/Pathway)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Results)
* [2014 Paris Bettencourt](https://2014.igem.org/Team%3AParis_Bettencourt/Results)