iGEM 2018 — Team SBS_SH_112144

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# Team:SBS SH 112144/Notebook

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* [Home](https://2018.igem.org/Team%3ASBS_SH_112144)
* [Project](https://2018.igem.org/Team%3ASBS_SH_112144/Description)
  + [Our Project](https://2018.igem.org/Team%3ASBS_SH_112144/Description)
  + [Intro to Lysozyme](https://2018.igem.org/Team%3ASBS_SH_112144/Lysozyme)
  + [Experiment Results](https://2018.igem.org/Team%3ASBS_SH_112144/Experiments)
  + [Notebook](https://2018.igem.org/Team%3ASBS_SH_112144/Notebook)
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* [Demo](https://2018.igem.org/Team%3ASBS_SH_112144/Demonstrate)
  + [Demonstrate](https://2018.igem.org/Team%3ASBS_SH_112144/Demonstrate)
  + [Device](https://2018.igem.org/Team%3ASBS_SH_112144/Device)
  + [Model](https://2018.igem.org/Team%3ASBS_SH_112144/Model)
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* [Modeling](https://2018.igem.org/Team%3ASBS_SH_112144/Model)
* [Human practice](https://2018.igem.org/Team%3ASBS_SH_112144/Human_Practices)
  + [Human Practice](https://2018.igem.org/Team%3ASBS_SH_112144/Human_Practices)
  + [Public Engagement](https://2018.igem.org/Team%3ASBS_SH_112144/Public_Engagement)
* [Parts](https://2018.igem.org/Team%3ASBS_SH_112144/Parts)
  + [Parts](https://2018.igem.org/Team%3ASBS_SH_112144/Parts)
  + [Improvement](https://2018.igem.org/Team%3ASBS_SH_112144/Improve)
  + [Basic Part](https://2018.igem.org/Team%3ASBS_SH_112144/Basic_Part)
  + [Composite Part](https://2018.igem.org/Team%3ASBS_SH_112144/Composite_Part)
* [Interlab](https://2018.igem.org/Team%3ASBS_SH_112144/InterLab)
* [Team](https://2018.igem.org/Team%3ASBS_SH_112144/Team)
  + [Team members](https://2018.igem.org/Team%3ASBS_SH_112144/Team)
  + [Collaborations](https://2018.igem.org/Team%3ASBS_SH_112144/Collaborations)
  + [Attribution](https://2018.igem.org/Team%3ASBS_SH_112144/Attributions)

#### Timeline

This timeline document is to keep track of our activities inside and outside the lab

[Timeline](https://static.igem.org/mediawiki/2018/9/99/T--SBS_SH_112144--Cyano1.pdf)

#### Lab Notes

This lab note is the primary source that would guide you through the majority of our experiments

[Lab Notes](https://static.igem.org/mediawiki/2018/e/e4/T--SBS_SH_112144--Cyano2.pdf)

#### General Protocol

This document General Protocol and Methods are for those who wish to learn how basic procedures of synthetic biology involved in our experiment works

[General Protocol](https://static.igem.org/mediawiki/2018/9/90/T--SBS_SH_112144--Cyano3.pdf)


[![](https://static.igem.org/mediawiki/2018/8/8b/HQ_page_logo.jpg)](https://2018.igem.org)

# Team:SBS SH 112144/Experiments

Header

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* [Home](https://2018.igem.org/Team%3ASBS_SH_112144)
* [Project](https://2018.igem.org/Team%3ASBS_SH_112144/Description)
  + [Our Project](https://2018.igem.org/Team%3ASBS_SH_112144/Description)
  + [Intro to Lysozyme](https://2018.igem.org/Team%3ASBS_SH_112144/Lysozyme)
  + [Experiment Results](https://2018.igem.org/Team%3ASBS_SH_112144/Experiments)
  + [Notebook](https://2018.igem.org/Team%3ASBS_SH_112144/Notebook)
  + [Reference](https://2018.igem.org/Team%3ASBS_SH_112144/Reference)
* [Demo](https://2018.igem.org/Team%3ASBS_SH_112144/Demonstrate)
  + [Demonstrate](https://2018.igem.org/Team%3ASBS_SH_112144/Demonstrate)
  + [Device](https://2018.igem.org/Team%3ASBS_SH_112144/Device)
  + [Model](https://2018.igem.org/Team%3ASBS_SH_112144/Model)
  + [Experiment results](https://2018.igem.org/Team%3ASBS_SH_112144/Experiments)
* [Modeling](https://2018.igem.org/Team%3ASBS_SH_112144/Model)
* [Human practice](https://2018.igem.org/Team%3ASBS_SH_112144/Human_Practices)
  + [Human Practice](https://2018.igem.org/Team%3ASBS_SH_112144/Human_Practices)
  + [Public Engagement](https://2018.igem.org/Team%3ASBS_SH_112144/Public_Engagement)
* [Parts](https://2018.igem.org/Team%3ASBS_SH_112144/Parts)
  + [Parts](https://2018.igem.org/Team%3ASBS_SH_112144/Parts)
  + [Improvement](https://2018.igem.org/Team%3ASBS_SH_112144/Improve)
  + [Basic Part](https://2018.igem.org/Team%3ASBS_SH_112144/Basic_Part)
  + [Composite Part](https://2018.igem.org/Team%3ASBS_SH_112144/Composite_Part)
* [Interlab](https://2018.igem.org/Team%3ASBS_SH_112144/InterLab)
* [Team](https://2018.igem.org/Team%3ASBS_SH_112144/Team)
  + [Team members](https://2018.igem.org/Team%3ASBS_SH_112144/Team)
  + [Collaborations](https://2018.igem.org/Team%3ASBS_SH_112144/Collaborations)
  + [Attribution](https://2018.igem.org/Team%3ASBS_SH_112144/Attributions)

![](https://static.igem.org/mediawiki/2018/c/c8/T--SBS_SH_112144--Experiment-results.png)

#### Molecular cloning

1. We were able to use PCR to successfully amplify the synthesized gene (gel unshown)

2. Through homologous recombination using an exonuclease kit, we were able to fuse the cp-OS lysozyme 1 gene into the PET28A and PSB1C3 plasmid

![](https://static.igem.org/mediawiki/2018/9/96/T--SBS_SH_112144--Cyanobacteria6.jpg)

3. We were able to cultivate the transformed BL21 E.coli and the addition of IPTG successfully triggered the expression of lysozyme.

#### Protein Purification

By purifying the protein using nickel column and later cleaving the His tag using TEV enzyme, we obtained pure lysozyme testified by the clearly labeled gel below.

![](https://static.igem.org/mediawiki/2018/8/82/T--SBS_SH_112144--Cyanobacteria8_.jpg)

Protein Acrylamide Gel on the purified lysozyme

#### Function test

1. By applying the lysozyme to a system（300ml in a microfuge tube reacting under different temperature) with bugbuster, cyanobacteria, pH buffer and water, we are able to observe cleavage effect illustrated by the OD660nm green light absorptance across large temperature and pH range. And through a series of mathematical computation and [modeling](https://2018.igem.org/Team%3ASBS_SH_112144/Model), we are able to identify that the ideal pH is around 6.9 and temperature is at 37 °C.

![](https://static.igem.org/mediawiki/2018/f/fb/T--SBS_SH_112144--Cyanobacteria10_.jpg)

Diagram we used to optimize pH and temperature

2. After finding the ideal pH and temperature for the lysozyme, we successfully applied these elements to a new set of function test with different protein concentration and reaction time, and we are also able to, through PSO algorithm and 3D modeling in matlab, find out the optimal reaction time being 0.5h( quite different from the paper) and the protein concentration being 18 ug/mL.

![](https://static.igem.org/mediawiki/2018/5/5d/T--SBS_SH_112144--Cyanobacteria11_.jpg)

Diagram we used to optimize enzyme concentration and reaction time

3. Lastly, we built the prototype of our [device](https://2018.igem.org/Team%3ASBS_SH_112144/Device) to test the effectiveness of our device under the provided parameters to roughly mimic the actual commercial operation of such a device. And we were thrilled to see an over 70-80 percent of lysis even in the prototype.

![](https://static.igem.org/mediawiki/2018/4/4a/T--SBS_SH_112144--Cyanobacteria12_.jpg)

![](https://static.igem.org/mediawiki/2018/b/b0/T--SBS_SH_112144--Cyanobacteria13_.jpg)

[Lab Notebook and Protocol](https://2018.igem.org/Team%3ASBS_SH_112144/Notebook%20)[Device](https://2018.igem.org/Team%3ASBS_SH_112144/Device)[Project Description](https://2018.igem.org/Team%3ASBS_SH_112144/Description)

[![](https://static.igem.org/mediawiki/2018/8/8b/HQ_page_logo.jpg)](https://2018.igem.org)

# Team:SBS SH 112144/Results

# Results

Here you can describe the results of your project and your future plans.

### 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 be on part pages in the Registry.
* 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)