Team:Goettingen/Team/Reporter

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<p><b>Figure 1a:</b> In the absence of c-di-AMP, DarR cannot bind to its binding site (operator) that is located downstream of the promoter. As a consequence the reporter gene is expressed and the cells synthesizing the green fluorescent protein (GFP) become fluorescent.</p>
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<p><b>Figure 1 A:</b> In the absence of c-di-AMP, DarR cannot bind to its binding site (operator) that is located downstream of the promoter. As a consequence the reporter gene is expressed and the cells synthesizing the green fluorescent protein (GFP) become fluorescent.</p>
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<p><b>Figure 1b: </b> In the presence of c-di-AMP, the signaling molecule stimulates DNA-binding activity of DarR and the c-di-AMP-DarR complex prevents transcription initiation by RNA polymerase. The <i>E. coli cells</i> do not produce GFP and are therefore non-fluorescent.</p></div>
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<p><b>Figure 1 B: </b> In the presence of c-di-AMP, the signaling molecule stimulates DNA-binding activity of DarR and the c-di-AMP-DarR complex prevents transcription initiation by RNA polymerase. The <i>E. coli</i> cells do not produce GFP and are therefore non-fluorescent.</p></div>
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<p>Reference:</p>
<p>Reference:</p>

Revision as of 13:59, 27 September 2013





The beast and its Achilles heel:

 A novel target to fight multi-resistant pathogenic bacteria



Reporter Team

The activity of transcriptional regulators can be easily detected by monitoring the activity of a promoter that is fused to a reporter gene. Recently, in Mycobacterium smegmatis, a transcriptional repressor (DarR) was identified that is able to bind to a specific DNA sequence upon association with c-di-AMP. This led us to the idea of developing a screening system for potential drugs interfering with c-di-AMP biofunction. While many Gram-positive bacteria require c-di-AMP for their growth, this molecule is not synthesized by the Gram-negative bacterium E. coli. Thus, we intend to build a reporter system that is composed of existing and of novel biobricks that will be developed during the project (Figure 1).


Reference:

1. Zhang et al. (2013) DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in Mycobacterium smegmatis, J. Biol. Chem. 288:3085–3096

 

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