iGEM 2019 — Team Hong_Kong_HKU

[![](https://2019.igem.org/wiki/images/8/8b/HQ_page_logo.jpg)](https://2019.igem.org)

# Team:Hong Kong HKU/Notebook

iGEM\_2019\_HKU\_Bootstrap\_Template

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[![](https://static.igem.org/mediawiki/2019/a/a1/T--Hong_Kong_HKU--bluelo.png)](https://2019.igem.org/Team%3AHong_Kong_HKU)

* People
  + [Attribution](https://2019.igem.org/Team%3AHong_Kong_HKU/Attributions)
  + [Members](https://2019.igem.org/Team%3AHong_Kong_HKU/Team)
* Project
  + [Design Overview](https://2019.igem.org/Team%3AHong_Kong_HKU/Description)
  + [Nanostructure Synthesis](https://2019.igem.org/Team%3AHong_Kong_HKU/Nanostructure_Synthesis)
  + [Co-culture Optimization](https://2019.igem.org/Team%3AHong_Kong_HKU/Co-culture_Optimization)
  + [Gene Delivery](https://2019.igem.org/Team%3AHong_Kong_HKU/Gene_Delivery)
  + [Modelling](https://2019.igem.org/Team%3AHong_Kong_HKU/Model)
* Parts
  + [Parts Overview](https://2019.igem.org/Team%3AHong_Kong_HKU/Parts)
  + [Parts Collection](https://2019.igem.org/Team%3AHong_Kong_HKU/Part_Collection)
  + [Characterization](https://2019.igem.org/Team%3AHong_Kong_HKU/Characterization)
* [Safety](https://2019.igem.org/Team%3AHong_Kong_HKU/Safety)
* Human Practices
  + [Integrated Human Practices](https://2019.igem.org/Team%3AHong_Kong_HKU/Human_Practices)
  + [Mentorship Collaboration](https://2019.igem.org/Team%3AHong_Kong_HKU/Collaborations)
  + [Education & Engagement](https://2019.igem.org/Team%3AHong_Kong_HKU/Public_Engagement)

* Presentation and Deliverables
  + [Presentation](https://2019.igem.org/Team%3AHong_Kong_HKU/Presentation)
  + [Poster](https://2019.igem.org/Team%3AHong_Kong_HKU/Poster)
  + [Team Video](https://2019.igem.org/Team%3AHong_Kong_HKU/TeamVideo)
* [Logbook](https://2019.igem.org/Team%3AHong_Kong_HKU/Notebook)
* [Judging](https://igem.org/2019_Judging_Form?id=3063)

## Logbook

##### **Sponsors**

![](https://static.igem.org/mediawiki/2017/a/a1/Blossom_%28Logo%29-19-5-2015.jpg)
![](https://static.igem.org/mediawiki/2017/2/2a/SnapGene_Logo_288HKU.png)
![](https://static.igem.org/mediawiki/2019/1/16/T--Hong_Kong_HKU--IDThhh.png)

---

##### **Contact us**

[![](https://static.igem.org/mediawiki/2019/archive/c/c2/20191020203121%21T--Hong_Kong_HKU--Foot_fb.png)](https://www.facebook.com/hkuigem2019/)
[![](https://static.igem.org/mediawiki/2019/d/d4/T--Hong_Kong_HKU--Foot_IG.png)](https://www.instagram.com/igem_hku/)

**Or contact us via Gmail ![](https://static.igem.org/mediawiki/2019/f/f4/T--Hong_Kong_HKU--Foot_Gmail.png): igemhku@hku.hk**


[![](https://2019.igem.org/wiki/images/8/8b/HQ_page_logo.jpg)](https://2019.igem.org)

# Team:Hong Kong HKU/Experiments

* [![Tetrahedron](https://static.igem.org/mediawiki/2019/a/a5/T--Hong_Kong_HKU--Logo.png)](https://2019.igem.org/Team%3AHong_Kong_HKU)
* [People](https://2019.igem.org/Team%3AHong_Kong_HKU/Attributions)
  + [Attributions](https://2019.igem.org/Team%3AHong_Kong_HKU/Attributions)
  + [Members](https://2019.igem.org/Team%3AHong_Kong_HKU/Team)* [Project](https://2019.igem.org/Team%3AHong_Kong_HKU/Description)
    + [Design Overview](https://2019.igem.org/Team%3AHong_Kong_HKU/Description)
    + [Experimental Results](https://2019.igem.org/Team%3AHong_Kong_HKU/Results)
    + [Demonstration](https://2019.igem.org/Team%3AHong_Kong_HKU/Demonstrate)
    + [Model](https://2019.igem.org/Team%3AHong_Kong_HKU/Model)* [Parts](https://2019.igem.org/Team%3AHong_Kong_HKU/Parts)
      + [Parts Overview](https://2019.igem.org/Team%3AHong_Kong_HKU/Parts)
      + [Part Collection](https://2019.igem.org/Team%3AHong_Kong_HKU/Part_Collection)
      + [Characterization](https://2019.igem.org/Team%3AHong_Kong_HKU/Characterization)* [Safety](https://2019.igem.org/Team%3AHong_Kong_HKU/Safety)
      * [Human Practices](https://2019.igem.org/Team%3AHong_Kong_HKU/Human_Practices)
        + [Integrated Human Practices](https://2019.igem.org/Team%3AHong_Kong_HKU/Human_Practices)
        + [Mentorship Collaboration](https://2019.igem.org/Team%3AHong_Kong_HKU/Collaborations)
        + [Education & Engagement](https://2019.igem.org/Team%3AHong_Kong_HKU/Public_Engagement)* [Logbook](https://2019.igem.org/Team%3AHong_Kong_HKU/Notebook)
        * [Judging](https://igem.org/2018_Judging_Form?team=Hong_Kong_HKU)

# 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.

Please remember to put all characterization and measurement data for your parts on the corresponding Registry part pages.

### What should this page contain?

* Protocols
* Experiments
* Documentation of the development of your project

### 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://2019.igem.org/wiki/images/8/8b/HQ_page_logo.jpg)](https://2019.igem.org)

# Team:Hong Kong HKU/Results

iGEM\_2019\_HKU\_Bootstrap\_Template

Toggle navigation

[![](https://static.igem.org/mediawiki/2019/a/a1/T--Hong_Kong_HKU--bluelo.png)](https://2019.igem.org/Team%3AHong_Kong_HKU)

* People
  + [Attribution](https://2019.igem.org/Team%3AHong_Kong_HKU/Attributions)
  + [Members](https://2019.igem.org/Team%3AHong_Kong_HKU/Team)
* Project
  + [Design Overview](https://2019.igem.org/Team%3AHong_Kong_HKU/Description)
  + [Nanostructure Synthesis](https://2019.igem.org/Team%3AHong_Kong_HKU/Nanostructure_Synthesis)
  + [Co-culture Optimization](https://2019.igem.org/Team%3AHong_Kong_HKU/Co-culture_Optimization)
  + [Gene Delivery](https://2019.igem.org/Team%3AHong_Kong_HKU/Gene_Delivery)
  + [Modelling](https://2019.igem.org/Team%3AHong_Kong_HKU/Model)
* Parts
  + [Parts Overview](https://2019.igem.org/Team%3AHong_Kong_HKU/Parts)
  + [Parts Collection](https://2019.igem.org/Team%3AHong_Kong_HKU/Part_Collection)
  + [Characterization](https://2019.igem.org/Team%3AHong_Kong_HKU/Characterization)
* [Safety](https://2019.igem.org/Team%3AHong_Kong_HKU/Safety)
* Human Practices
  + [Integrated Human Practices](https://2019.igem.org/Team%3AHong_Kong_HKU/Human_Practices)
  + [Mentorship Collaboration](https://2019.igem.org/Team%3AHong_Kong_HKU/Collaborations)
  + [Education & Engagement](https://2019.igem.org/Team%3AHong_Kong_HKU/Public_Engagement)

* Presentation and Deliverables
  + [Presentation](https://2019.igem.org/Team%3AHong_Kong_HKU/Presentation)
  + [Poster](https://2019.igem.org/Team%3AHong_Kong_HKU/Poster)
  + [Team Video](https://2019.igem.org/Team%3AHong_Kong_HKU/TeamVideo)
* [Logbook](https://2019.igem.org/Team%3AHong_Kong_HKU/Notebook)
* [Judging](https://igem.org/2019_Judging_Form?id=3063)

# Experimental Results

### **DNA Nanostructures**

### Structural Characterisation

![](https://static.igem.org/mediawiki/2019/5/5a/T--Hong_Kong_HKU--tetra_page_2019.png)

The sizes of the 4 ssDNA component strands are shown in lanes 2-5. We confirmed the successful annealing of our DNA tetrahedron on lanes 6-8 in a stepwise manner when adding up ssDNA.

### Transmission Electron Microscopy

![](https://static.igem.org/mediawiki/2019/4/4f/T--Hong_Kong_HKU--TEM_2019.png)

Our DNA tetrahedral structures are stained and visualised by a Philips CM 100 Transmission Electron Microscope with 100 kV operating voltage.

Since the 3' end of each strand is modified to have a thiol group, gold nanoparticles could then be attached to it. After the strands assemble, all four vertices of the final tetrahedron will be attached by a gold nanoparticle and could be observed under the Transmission Electron Microscope.

### **Flagellar Over-expression**

![](https://static.igem.org/mediawiki/2019/2/2b/T--Hong_Kong_HKU--flhDCPCR1.jpg)

To drive the over-expression of flagella in Salmonella Typhimurium, we constructed two BioBricks: 1) flagella transcription factor (flhDC) and terminator; 2) flagella transcription factor with both the *lac* promoter and terminator.

To test whether we successfully cloned the constructs into the vector, colony PCR was done on 2 rounds of sub-cloning. First, the sequence encoding flhDC was inserted into pSB1C3. Secondly, the fragment containing both the flhDC and terminator was cut and inserted into the final vector containing *lac* promoter. The results of colony PCR show the successful cloning of two BioBricks.

Lanes 2-4 are triplicates of the correct constructs containing the *lac* promoter, flhDC and a terminator with matching sizes (1582bp). Lane 5 contains the PCR product of the intermediate constructs with only the flhDC sequence and a terminator with matching sizes (1382bp).

![](https://static.igem.org/mediawiki/2019/7/71/T--Hong_Kong_HKU--Flagellar.png)

To test whether the construct works and its effect, the BioBrick containing the *lac* promoter, flagella transcription factor flhDC sequence and terminator was transformed to Salmonella Typhimurium.

The transformed S. Typhimurium were then cultured inside a liver cancer cell spheroid of Huh7 cell line. After incubation, the content of the spheroid were diluted and spread on a plate for Colony Forming Units (CFU) measurement.

The results show that the transformed S. Typhimurium which expressed a lot more flagella form much more colonies than the normal S. Typhimurium. Because increased number of flagella will increase the motility of Salmonella and thus they can move faster and form more colonies.

##### **Sponsors**

![](https://static.igem.org/mediawiki/2017/a/a1/Blossom_%28Logo%29-19-5-2015.jpg)
![](https://static.igem.org/mediawiki/2017/2/2a/SnapGene_Logo_288HKU.png)
![](https://static.igem.org/mediawiki/2019/1/16/T--Hong_Kong_HKU--IDThhh.png)

---

##### **Contact us**

[![](https://static.igem.org/mediawiki/2019/archive/c/c2/20191020203121%21T--Hong_Kong_HKU--Foot_fb.png)](https://www.facebook.com/hkuigem2019/)
[![](https://static.igem.org/mediawiki/2019/d/d4/T--Hong_Kong_HKU--Foot_IG.png)](https://www.instagram.com/igem_hku/)

**Or contact us via Gmail ![](https://static.igem.org/mediawiki/2019/f/f4/T--Hong_Kong_HKU--Foot_Gmail.png): igemhku@hku.hk**