iGEM 2010 — Team Bielefeld-Germany

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# Team:Bielefeld-Germany/Notebook

### From 2010.igem.org

This is a template page. READ THESE INSTRUCTIONS.

You are provided with this team page template with which to start the iGEM season. You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples [HERE](https://2008.igem.org/Help%3ATemplate/Examples).

You **MUST** have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page. PLEASE keep all of your pages within your teams namespace.

|  |  |
| --- | --- |
| You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing. | [File:Bielefeld-Germany logo.png](/File%3ABielefeld-Germany_logo.png "File:Bielefeld-Germany logo.png") 200px |
| *Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs)* | [File:Bielefeld-Germany team.png](/File%3ABielefeld-Germany_team.png "File:Bielefeld-Germany team.png") Your team picture |
|  | [Team Example](/Team%3ABielefeld-Germany "Team:Bielefeld-Germany") |

| [Home](/Team%3ABielefeld-Germany "Team:Bielefeld-Germany") | [Team](/Team%3ABielefeld-Germany/Team "Team:Bielefeld-Germany/Team") | [Official Team Profile](https://igem.org/Team.cgi?year=2010&team_name=Bielefeld-Germany) | [Project](/Team%3ABielefeld-Germany/Project "Team:Bielefeld-Germany/Project") | [Parts Submitted to the Registry](/Team%3ABielefeld-Germany/Parts "Team:Bielefeld-Germany/Parts") | [Modeling](/Team%3ABielefeld-Germany/Modeling "Team:Bielefeld-Germany/Modeling") | **Notebook** | [Safety](/Team%3ABielefeld-Germany/Safety "Team:Bielefeld-Germany/Safety") |
| --- | --- | --- | --- | --- | --- | --- | --- |

## Notebook

You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.

Retrieved from "[http://2010.igem.org/Team:Bielefeld-Germany/Notebook](http://2010.igem.org/Team%3ABielefeld-Germany/Notebook)"

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# Team:Bielefeld-Germany/Results

### From 2010.igem.org

[![](https://static.igem.org/mediawiki/2010/3/3f/IGEM_basic_Logo_mini.png)](https://2010.igem.org)

![iGEM-Bielefeld](https://static.igem.org/mediawiki/2010/1/14/Bielefeld_banner_team.jpg)
[![](https://static.igem.org/mediawiki/2010/f/fe/Bielefeld_logo_small_black_dropshadow.png)](/Team%3ABielefeld-Germany)

- [Start](https://2010.igem.org/Team%3ABielefeld-Germany)

- [Team](https://2010.igem.org/Team%3ABielefeld-Germany/Team)

* [Description](https://2010.igem.org/Team%3ABielefeld-Germany/Team/Description)
* [Supervisors](https://2010.igem.org/Team%3ABielefeld-Germany/Team/Supervisors)
* [University](https://2010.igem.org/Team%3ABielefeld-Germany/Team/Bielefeld)
* [Bielefeld](https://2010.igem.org/Team%3ABielefeld-Germany/Team/BielefeldCity)
* [Motivation](https://2010.igem.org/Team%3ABielefeld-Germany/Team/Motivation)
* [Acknowledgements](https://2010.igem.org/Team%3ABielefeld-Germany/Team/Acknowledgements)

- [Project](https://2010.igem.org/Team%3ABielefeld-Germany/Project)

* [Application](https://2010.igem.org/Team%3ABielefeld-Germany/Project/Application)
* [Approach](https://2010.igem.org/Team%3ABielefeld-Germany/Project/Approach)
* [Theory](https://2010.igem.org/Team%3ABielefeld-Germany/Project/Theory)
* [Protocols](https://2010.igem.org/Team%3ABielefeld-Germany/Project/Protocols)
* [Model](https://2010.igem.org/Team%3ABielefeld-Germany/Project/Model)
* [Outlook](https://2010.igem.org/Team%3ABielefeld-Germany/Project/Outlook)

- [Results](https://2010.igem.org/Team%3ABielefeld-Germany/Results)

* [Characterization](https://2010.igem.org/Team%3ABielefeld-Germany/Results/Characterization)
* [Used BBs](https://2010.igem.org/Team%3ABielefeld-Germany/Results/Used)
* [Submitted BBs](https://2010.igem.org/Team%3ABielefeld-Germany/Results/Submitted)
* [Sequencing](https://2010.igem.org/Team%3ABielefeld-Germany/Results/Sequencing)

- [Safety](https://2010.igem.org/Team%3ABielefeld-Germany/Safety)

* [Biosecurity](https://2010.igem.org/Team%3ABielefeld-Germany/Safety#Biosecurity)
* [Biosafety](https://2010.igem.org/Team%3ABielefeld-Germany/Safety#Biosafety)
* [Risk Assesement](https://2010.igem.org/Team%3ABielefeld-Germany/Safety#Risk_Assesement_.28english.29)

- [Public Relations](https://2010.igem.org/Team%3ABielefeld-Germany/Public_Relations)

* [Fotos](https://2010.igem.org/Team%3ABielefeld-Germany/Public_Relations/Fotos)
* [Press](https://2010.igem.org/Team%3ABielefeld-Germany/Public_Relations/Press)
* [Public Discussion](https://2010.igem.org/Team%3ABielefeld-Germany/Public_Relations/Public_Discussion)

- [Judging](https://2010.igem.org/Team%3ABielefeld-Germany/Judging)

* [BioBricks](https://2010.igem.org/Team%3ABielefeld-Germany/Judging/BioBricks)
* [Helping other teams](https://2010.igem.org/Team%3ABielefeld-Germany/Judging/Helping)
* [Comments on iGEM](https://2010.igem.org/Team%3ABielefeld-Germany/Judging/Comments)

- [Sponsors](https://2010.igem.org/Team%3ABielefeld-Germany/Sponsors)

- [Contact](https://2010.igem.org/Team%3ABielefeld-Germany/Contact)

MENU

http://igem-bielefeld.de/img/banner\_lab.png

- [Results](/Team%3ABielefeld-Germany/Results)
- [Characterization](/Team%3ABielefeld-Germany/Results/Characterization)
- [Used](/Team%3ABielefeld-Germany/Results/Used)
- [Submitted](/Team%3ABielefeld-Germany/Results/Submitted)
- [Sequencing](/Team%3ABielefeld-Germany/Results/Sequencing)

# Results

[![](/wiki/images/0/02/Bielefeld_luc.jpg)](/File%3ABielefeld_luc.jpg)

[![](/wiki/skins/common/images/magnify-clip.png)](/File%3ABielefeld_luc.jpg "Enlarge")

**Figure 1: Visualization of induced (from left to right) <partinfo>K389421</partinfo>, <partinfo>K389422</partinfo> and <partinfo>K389423</partinfo> sensitivity tuner amplified vir-system.**

On the next pages we are going to present you our BioBrick work.

One approach of our project was to visualize a readout by a light emitting system. So a luciferase BioBrick was built. This BioBrick is a highly sensitive reporter gene which was successfully applied in different devices with a PoPS output. The characterization of this BioBrick can be found  [here](/Team%3ABielefeld-Germany/Results/Characterization/K389004 "Team:Bielefeld-Germany/Results/Characterization/K389004").

The second step was to establish a complete VirA/G signaling system from *Agrobacterium tumefaciens* in *Escherichia coli*. For this purpose three new BioBricks were created - *virA*, *virG* and a *vir* promoter. The VirA receptor recognizes the phenolic substance acetosyringone and transmits this information to the VirG response regulator - a bacterial transcription factor which activates *vir* promoters. This natural VirA/G signaling system works in *E. coli* and was measured and characterized by using the reporter genes  [mRFP](/Team%3ABielefeld-Germany/Results/Characterization/K389016 "Team:Bielefeld-Germany/Results/Characterization/K389016") and  [luciferase](/Team%3ABielefeld-Germany/Results/Characterization/K389015 "Team:Bielefeld-Germany/Results/Characterization/K389015").

In the third step we tried to modify the VirA receptor by directed mutagenesis using error-prone PCR, in order to detect substances other than acetosyringone (e.g. capsaicin). For that purpose we designed a  [high-throughput screening system](/Team%3ABielefeld-Germany/Project/Theory#Screening_system "Team:Bielefeld-Germany/Project/Theory") with a selective system based on the expression of a kanamycin resistance. We tried to modify the receptor by different modification strategies like error prone (random mutagenesis) or specific amino acid changes ([mut1 / mut2](/Team%3ABielefeld-Germany/Project/Approach#Directed_mutagenesis "Team:Bielefeld-Germany/Project/Approach")). The constructs mut1 and mut2 were done by an *in silico* approach. Unfortunately they did not work as expected. In a short period of time we found many positive clones, but further analysis indicated that the VirA receptor was changed to constitutive activity. Thereby, due to the short time available, we did not manage to construct a receptor that was specific for other phenolic substance, e.g. capsaicin.

We tried to create a light signal that is visible to the human eye as another part of our project. So genetic amplifiers ([Cambridge, iGEM 2007, amplifier project](https://2007.igem.org/Cambridge/Amplifier_project#Results)) were added to our luciferase BioBrick. This amplified luciferase readout was coupled with the VirA/G signaling system in order to visualize the induction behaviour. The amplifiers worked so far and gave a visible luciferase readout but the basal expression of the *vir* promoter was also amplified so the induced status of the VirA/G system was not discriminable properly from the uninduced status.

We entered the following BioBricks to the partsregistry:

<groupparts>iGEM010 Bielefeld-Germany</groupparts>

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