iGEM 2021 — Team Worldshaper-Nanjing

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

# Team:Worldshaper-Nanjing/Notebook

![](https://static.igem.org/mediawiki/2021/e/e7/T--Worldshaper-Nanjing--Logo4.png)

* [Home](https://2021.igem.org/Team%3AWorldshaper-Nanjing)
* Team

  [Team Members](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Team)
  [Collaborations](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Collaborations)
  [Attributions](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Attributions)
* Project

  [Description](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Description)
  [Design](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Design)
  [Implementation](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Implementation)
  [Contribution](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Contribution)
* [Model](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Model)
* [Parts](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Parts)
* Experiment

  [Engineering
  Success](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Engineering)
  [Proof of
  Concept](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Proof_Of_Concept)
  [Protocols](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Protocols)
  [Notebook](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Notebook)
  [Safety](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Safety)
* Human Practices

  [Integrated Human
  Practices](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Human_Practices)
  [Education&Communication](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Communication)
  [Partnership](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Partnership)

# Notebook

[Home](https://2021.igem.org/Team%3AWorldshaper-Nanjing)
 >
Experiment
 >
[Notebook](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Notebook)

### April

* - iGEM competition rules learning
* - Professional theory learning
* - Projects brainstorming

Soon after the team was established, the team members were informed iGEM rules
comprehensively and started to learn some related theories.
We studied the wikis of previous iGEM teams and meanwhile brainstormed for our own topic.

### May

* - Find the project goals
* - Background survey
* - Professional theory learning
* - Group division
* - Work schedule

Taking utility and feasibility into consideration, our team finally decided the topic and
began some background survey. At the same time, we set up several groups in charge of different tasks,
and each of the teammates, based on their personal interests and abilities, joined three of them.

### June

* - Team registration
* - Background survey
* - Background manuscript
* - Lab safety training
* - Self-media update
* - Team logo design
* - Team leader selection
* - Interview plan

We finished our background survey and wrote a manuscript. Since in a month we would enter the lab,
we learnt more about lab safety and prepared all the things required in the following months.
The team also worked out an interview plan,
which would help us learn more about gutter oil from different perspectives.

### July

#### Wet Team

* - Project theory and experimental skills
* - Daily experiment record
* - Plasmids constructing and transforming

#### Dry Team

* - Promotion video
* - Self-media update
* - Questionnaire collection
* - Academic exchange and cooperation activities
* - Some interviews
* - Expert interviews
* - Public survey
* - Partnership and collaboration

The team first learnt the experimental procedures. In the rest of the month,
the team successfully constructed nine plasmids by Gibson, spliced the mfe1-pot1 plasmid,
transformed it into yeast and introduced the yeast into the medium.
The dry team also carried on their work during the month. We distributed a questionnaire to figure out the public’s attitudes,
and interviewed some professionals to know more about the current situation.

### August

#### Wet Team

* - The experiment was suspended because of the epidemic situation
* - Further project design
* - Project description
* - Protocols
* - Fermentor design

#### Dry Team

* - Self-media update
* - Online education activities
* - Picture book
* - Collaboration
* - Human practice
* - Questionnaire collection
* - Online meetups

Unfortunately, due to the outbreak of Covid-19 in Nanjing, the experiment was suspended according to the local policy.
However, at the same time it provided us with abundant time for other work. We continued our collaboration plan,
including holding online education activities and finishing a propagandistic picture book with other iGEM teams.
After collecting the data of the questionnaire, we analyzed the result and came to some conclusions.

### September&October

* - Complete the experiment
* - Wiki manuscript
* - Modeling
* - Applied design
* - Prepare for presentation
* - Presentation video
* - Poster
* - Wiki web page optimization
* - Final judging form
* - Parts registry

# Worldshaper-Nanjing 2021

![](https://static.igem.org/mediawiki/2021/e/e7/T--Worldshaper-Nanjing--Logo4.png)

## Contact Us

* Wechat: NJ Shaper

## Links

* [Home](https://2021.igem.org/Team%3AWorldshaper-Nanjing)
* [Team](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Team)
* [Project](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Description)
* [Model](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Model)
* [Parts](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Parts)
* [Experiment](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Engineering)
* [Human Practice](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Human_Practices)

## QR Code

![](https://static.igem.org/mediawiki/2021/c/c1/T--Worldshaper-Nanjing--WechatQRCodeTest04.png)

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

# Team:Worldshaper-Nanjing/Engineering

![](https://static.igem.org/mediawiki/2021/e/e7/T--Worldshaper-Nanjing--Logo4.png)

* [Home](https://2021.igem.org/Team%3AWorldshaper-Nanjing)
* Team

  [Team Members](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Team)
  [Collaborations](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Collaborations)
  [Attributions](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Attributions)
* Project

  [Description](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Description)
  [Design](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Design)
  [Implementation](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Implementation)
  [Contribution](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Contribution)
* [Model](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Model)
* [Parts](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Parts)
* Experiment

  [Engineering
  Success](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Engineering)
  [Proof of
  Concept](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Proof_Of_Concept)
  [Protocols](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Protocols)
  [Notebook](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Notebook)
  [Safety](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Safety)
* Human Practices

  [Integrated Human
  Practices](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Human_Practices)
  [Education&Communication](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Communication)
  [Partnership](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Partnership)

# Engineering Success

[Home](https://2021.igem.org/Team%3AWorldshaper-Nanjing)
 >
Project
 >
[Engineering Success](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Engineering)

### 1 Overview

---

Y. lipolytica, as a ‘generally regarded as safe’ (GRAS) yeast, has been extensively engineered for the production of oleochemicals, fuels and commodity chemicals. In this project, to achieve the production of γ-linolenic acid with the gutter oil as substrate, we introduced the Δ6-fatty acid desaturase and overexpressed the β-oxidation pathway in the Y. lipolytica po1f. For this, we firstly constructed the engineering stains with the strengthened β-oxidation pathway by the combined overexpression of genes ylPOT1, ylMFE1, and ylPOX1-6. The experimental results showed that combined overexpression of genes ylPOT1, ylMFE1, and ylPOX4 or ylPOX5 significantly improve the cell growth of Y. lipolytica with using the edible oil as the substrate. Next, we introduced the Δ6-fatty acid desaturase encoding gene M-Δ6-D into the engineered strains with improved cell growth by the genomic integration method. As a result, we obtained a titer of 59.3 mg/L γ-linolenic acid from the engineered strain with using the gutter oil as substrate. The result set up a new stage of engineering Y. lipolytica as a sustainable biorefinery chassis strain for the synthesis of γ-linolenic acid with the gutter oil as substrate.

### 2 Engineering success

---

#### 2.1 Constructing recombinant plasmids for overexpressing the β-oxidation pathway

**2.1.1 PCR amplification of genes**

Firstly, we amplified the sequences of genes ylPOT1, ylMFE1, ylPOX1, ylPOX2, ylPOX3, ylPOX4, ylPOX5 , and ylPOX6 with using the Y. lipolytica as the template by PCR method. The result of genes amplification has been showed in Figure 1.

![](https://static.igem.org/mediawiki/2021/3/32/T--Worldshaper-Nanjing--EngineeringSuccess01.png)

# Worldshaper-Nanjing 2021

![](https://static.igem.org/mediawiki/2021/e/e7/T--Worldshaper-Nanjing--Logo4.png)

## Contact Us

* Wechat: NJ Shaper

## Links

* [Home](https://2021.igem.org/Team%3AWorldshaper-Nanjing)
* [Team](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Team)
* [Project](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Description)
* [Model](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Model)
* [Parts](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Parts)
* [Experiment](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Engineering)
* [Human Practice](https://2021.igem.org/Team%3AWorldshaper-Nanjing/Human_Practices)

## QR Code

![](https://static.igem.org/mediawiki/2021/c/c1/T--Worldshaper-Nanjing--WechatQRCodeTest04.png)

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

# Team:Worldshaper-Nanjing/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:Worldshaper-Nanjing/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)