Team:Goettingen/Project/OurProject

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<p>Our project is aimed at the development of a simple screening system, which allows the rapid identification and characterization of substances that disturb c-di-AMP homeostasis in pathogenic bacteria. By developing this system, we believe we can help scientist and pharmaceutical companies worldwide to find new and more effective antibiotics against Gram-positive pathogens, for which c-di-AMP is essential. The screening system will be established in the non- pathogenic bacterium <i>E. coli.</i> </p>
<p>Our project is aimed at the development of a simple screening system, which allows the rapid identification and characterization of substances that disturb c-di-AMP homeostasis in pathogenic bacteria. By developing this system, we believe we can help scientist and pharmaceutical companies worldwide to find new and more effective antibiotics against Gram-positive pathogens, for which c-di-AMP is essential. The screening system will be established in the non- pathogenic bacterium <i>E. coli.</i> </p>
<p>Using given Biobricks and by creating new ones, we first wanted to construct a promoter-reporter fusion system which is under the control of DarR, an operator derived from <i>Mycobacterium smegmatis</i>. Only in the presents of c-di-AMP will this operator able to inhibit the transcription of the reporter gene GFP. The operator binding sequence will be placed between a constitutively active and the reporter gene. The activity of this reporter is rather to detect, since it emits green light if not inhibited. Via the addition of exogenous c-di-AMP, we will have to evaluate whether it is able to inhibit our promoter-reporter system.</p>
<p>Using given Biobricks and by creating new ones, we first wanted to construct a promoter-reporter fusion system which is under the control of DarR, an operator derived from <i>Mycobacterium smegmatis</i>. Only in the presents of c-di-AMP will this operator able to inhibit the transcription of the reporter gene GFP. The operator binding sequence will be placed between a constitutively active and the reporter gene. The activity of this reporter is rather to detect, since it emits green light if not inhibited. Via the addition of exogenous c-di-AMP, we will have to evaluate whether it is able to inhibit our promoter-reporter system.</p>
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<img src="https://static.igem.org/mediawiki/2013/6/65/Goe-reporter-1.png" class="fl" style="display-inline;width:49%" />
<img src="https://static.igem.org/mediawiki/2013/c/c5/Goe-reporter-2.png" class="fl" style="display-inline;width:49%" />
<img src="https://static.igem.org/mediawiki/2013/c/c5/Goe-reporter-2.png" class="fl" style="display-inline;width:49%" />
<p>By accident we stumbled upon the discovery of a new Ribo-Switch, which is also under the control of c-di-AMP. This Ribo-Switch, called YdaO, was discovered in Bacillus subitilis.With this, we figured, we would have a second way to construct a reporter system. (Green Coli with a SWITCH in his hand)</p>
<p>By accident we stumbled upon the discovery of a new Ribo-Switch, which is also under the control of c-di-AMP. This Ribo-Switch, called YdaO, was discovered in Bacillus subitilis.With this, we figured, we would have a second way to construct a reporter system. (Green Coli with a SWITCH in his hand)</p>

Revision as of 14:11, 26 September 2013





The beast and its Achilles heel:

 A novel target to fight multi-resistant pathogenic bacteria



Our Porject

Our project is aimed at the development of a simple screening system, which allows the rapid identification and characterization of substances that disturb c-di-AMP homeostasis in pathogenic bacteria. By developing this system, we believe we can help scientist and pharmaceutical companies worldwide to find new and more effective antibiotics against Gram-positive pathogens, for which c-di-AMP is essential. The screening system will be established in the non- pathogenic bacterium E. coli.

Using given Biobricks and by creating new ones, we first wanted to construct a promoter-reporter fusion system which is under the control of DarR, an operator derived from Mycobacterium smegmatis. Only in the presents of c-di-AMP will this operator able to inhibit the transcription of the reporter gene GFP. The operator binding sequence will be placed between a constitutively active and the reporter gene. The activity of this reporter is rather to detect, since it emits green light if not inhibited. Via the addition of exogenous c-di-AMP, we will have to evaluate whether it is able to inhibit our promoter-reporter system.

By accident we stumbled upon the discovery of a new Ribo-Switch, which is also under the control of c-di-AMP. This Ribo-Switch, called YdaO, was discovered in Bacillus subitilis.With this, we figured, we would have a second way to construct a reporter system. (Green Coli with a SWITCH in his hand)

 

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