Team:Goettingen/Team/Reporter

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===Reporter Team===
===Reporter Team===
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==Introduction==
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<p>The activity of transcriptional regulators can be easily monitored by expressing a reporter gene downstream of a promoter, containing their binding site. 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 consisting of the iGEM biobricks in E. coli (Figure 1).</p>
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In order to facilitate the development of new antibiotics, we want to build an “Antibiotic Detector” that can identify substances influencing the homeostasis of cyclic-di-AMP (c-di-AMP). c-di-AMP is an essential signal nucleotide in most Gram-positive bacteria. It is not found in humas and several Gram-negative bacteria lack this compound, as well, including <i>E. coli</i>. For more information see [[Team:Goettingen/Project/OurProject|Our Project overview]].
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<img src="https://static.igem.org/mediawiki/2013/6/65/Goe-reporter-1.png" style="width:70%;display:block" class="cen" />
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For constructing a c-di-AMP detector, we need sensors for c-di-AMP and in order to visualize the output of the sensors, we need a reporter.  
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<div style="width:70%" class="cen"><b>Figure 1a:</b> When there is no c-di-AMP present, the repressor gene DarR cannot bind to its binding site(operator sequence). The transciption starts and leads to the expression of the reporter gene. This reporter gene could be GFP, or other widely used reporter gene.</div>
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==DarR reporter system ([http://parts.igem.org/Part:BBa_K1045017|part BBa_K1045017]):==
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<img src="https://static.igem.org/mediawiki/2013/c/c5/Goe-reporter-2.png" style="width:70%;display:block" class="cen" />
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<b>Figure 1b: </b> When there is c-di-AMP present, the c-di-AMP can act as ligand of the repressor gene DarR, activating tis function. DarR binds to its binding site, blocking the RNA polymerase, and therefore disrupt transcription. Then we are supposed to see no signal or reduced signal.</div>
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<p>Reference:</p>
<p>Reference:</p>
1. <a href="http://www.jbc.org/content/288/5/3085" style="color:#303030" >Zhang <i>et.</i>al.(2013)DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in <i>Mycobacterium smegmatis, J. Biol. Chem. </i> 288:3085–3096</a>
1. <a href="http://www.jbc.org/content/288/5/3085" style="color:#303030" >Zhang <i>et.</i>al.(2013)DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in <i>Mycobacterium smegmatis, J. Biol. Chem. </i> 288:3085–3096</a>
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Revision as of 09:13, 1 October 2013





The beast and its Achilles heel:

 A novel target to fight multi-resistant pathogenic bacteria



Reporter Team

Introduction

In order to facilitate the development of new antibiotics, we want to build an “Antibiotic Detector” that can identify substances influencing the homeostasis of cyclic-di-AMP (c-di-AMP). c-di-AMP is an essential signal nucleotide in most Gram-positive bacteria. It is not found in humas and several Gram-negative bacteria lack this compound, as well, including E. coli. For more information see Our Project overview.

For constructing a c-di-AMP detector, we need sensors for c-di-AMP and in order to visualize the output of the sensors, we need a reporter.

DarR reporter system ([http://parts.igem.org/Part:BBa_K1045017|part BBa_K1045017]):

Reference:

1. <a href="http://www.jbc.org/content/288/5/3085" style="color:#303030" >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</a>


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