iGEM 2024 — Team hnu-china

{% extends "layout.html" %}
{% block title %}Notebook{% endblock %}
{% block lead %}Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.{% endblock %}
{% block page\_content %}

![Background image](https://static.igem.wiki/teams/5116/background3.png)

## Useful Links

* [记录1](https://static.igem.wiki/teams/5116/amamamam.pdf)
* [记录2](https://static.igem.wiki/teams/5116/shiyanjilu.pdf)
* [记录3](https://static.igem.wiki/teams/5116/lalalala.pdf)

{% endblock %}

{% extends "layout.html" %}
{% block title %}Results{% endblock %}
{% block lead %}You can describe the results of your project and your future plans here.{% endblock %}
{% block page\_content %}

![img](https://static.igem.wiki/teams/5116/background3.png)

## preparation

---

As a beginner to work with pseudomonas, we made a lot attempts to culture and prepare workable P.F.strain.
So we tried culture medium with different ingredient. We discovered that it can grow on LBmedium ,but it will grow better on YPDA. In regular LBmedium, the max od600 will around 0.9 ,and in YPDA the max od600 will rise to around 1.4,which significantly rises.
Then we made some attempts to make competent P.F.cells. Both electroporation method and CaCl method were tried and we have success in CaCl method. Those competent P.F. will be used in further experiments.
We also tried to make culture curve of pseudomonas. 12 culture tubes were prepared and placed in the incubator with a gap of 1hours. We took them out every 12 hrs to record th od600.
We tested our plasmid extraction kit on P.F. and the result turns out that the regular plasmid extraction kit is not suitable for P.F. latter plasmids will be amplified in E.coli.
We tested several expression system that we can acquire from our PIs. Mainly pET28a, pGEX6p and pMR\* system. The system with pET28a backbone works really well. While pGEX6p will strongly inhibit the growth of P.F. and pMR\* system is not working at all. However the T7 promoter will not work properly in PF0-1 strain. So we have to re place the promoter.

## Plasmid design on P.F.

---

In our previous experiments we chose pET28a as our vector. The vector introduces a lactose operating system . the plasmid should contain two features:encodes a protein for lactose intake and the anti fungal peptide we use. The expression of the peptide should be controlled by lactose concentration. So we first build a artificial inducible promoter with constitutive promoter PpltR, which mentioned in team SKLMT 2018, and a Lac operator. The antifungal peptide coding with signal peptide assembled toit were atacched to such artificial promoter. Secondly,we use another gene LacY to ensure lactose intake.

## Plasmid design on yeast.

---

We use a dual vector system to make protein expression in yeast: pGADT7 and pGBKT7.
In pGADT7 we insert B4GalT1 gene with CYC1 promoter. It encodes β-1,4 udp-galactosyltransferase for lactose synthesis.
In pGBKT7 we insert LALBA gene with CYC1 promoter as well,it encodes lactalbuminα as it can enhance the specificity of B4GalT1
We will also insert GalE gene with FIG1 promoter. It will produce UGP-glucose epimerase when fungal hormone receptor is activated.

{% endblock %}

{% extends "layout.html" %}
{% block title %}Experiments{% endblock %}
{% block lead %}Describe the research, experiments, and protocols you used in your iGEM project.{% endblock %}
{% block page\_content %}

![Project background image](https://static.igem.wiki/teams/5116/background3.png)

## What should this page contain?

---

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](https://parts.igem.org/Main_Page).

## Inspirations

---

* [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)

{% endblock %}

{% extends "layout.html" %}
{% block title %}Results{% endblock %}
{% block lead %}You can describe the results of your project and your future plans here.{% endblock %}
{% block page\_content %}

![img](https://static.igem.wiki/teams/5116/background3.png)

## 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](https://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 and what you learned from them. 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. Furthermore, you can still impress the judges by thoughtfully discussing what went wrong and how you might change things.

## Inspirations

---

* [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)

{% endblock %}