Team:Goettingen/Team/Array

From 2013.igem.org

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===Array Team===
===Array Team===
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<p>By using the microarray technique we would like to analyze how many genes are regulated by ci-di-AMP in the Gram-positive model bacterium <i>Bacillus subtilis</i>. We are not only interested in the impact of c-di-AMP on the transcriptome, we also hope to identify novel regulatory elements (i.e. riboswitches) that bind to the signaling molecule. In our experiments we use three different <i>B. subtilis</i> strains. The wild-type strain contains all three diadenylatecyclases DisA, CdaA, CdaS. The second strain is a <i>disA</i> mutant strain lacking DisA, which was shown to be involved in DNA metabolism. This strain should produce less c-di-AMP than the isogenic parent strain. The third strain synthesizes a hyperactive CdaS mutant variant, which produces a lot of c-di-AMP. In our microarray experiments we will compare the transcriptomes of each mutant strain with that of the wild-type strain. Moreover, we would like to create a triple knock-out mutant (<i>ΔdisA, ΔcdaA, ΔcdaS</i>) lacking all diadenylate cyclases. We try to construct the triple knock-out mutant by feeding with exogenous c-di-AMP. </p>
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<p>By using the microarray technique we would like to analyze how many genes are regulated by ci-di-AMP in the Gram-positive model bacterium <i>Bacillus subtilis</i>. We are not only interested in the impact of c-di-AMP on the transcriptome, we also hope to identify novel regulatory elements (i.e. riboswitches) that bind to the signaling molecule. In our experiments we use three different <i>B. subtilis</i> strains. The wild-type strain contains all three diadenylatecyclases (DisA, CdaA, and CdaS). The second strain is a <i>disA</i> mutant strain lacking DisA, which was shown to be involved in DNA metabolism. This strain should produce less c-di-AMP than the isogenic parent strain. The third strain synthesizes a hyperactive CdaS mutant variant, which produces a lot of c-di-AMP. In our microarray experiments we will compare the transcriptomes of each mutant strain with that of the wild-type strain. Moreover, we would like to create a triple knock-out mutant (<i>ΔdisA, ΔcdaA, ΔcdaS</i>) lacking all diadenylate cyclases. We try to construct the triple knock-out mutant by feeding with exogenous c-di-AMP. </p>
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<p>Reference:</p>
<p>Reference:</p>
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1. <a href="http://www.jbc.org/content/288/5/3085" style="color:#303030" >Witte <i>et al.</i> (2008) Structural Biochemistry of a Bacterial Checkpoint Protein Reveals Diadenylate Cyclase Activity Regulated by DNA Recombination Intermediates. Mol. Cell. 30:167-178</a>
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1. Witte <i>et al.</i> (2008) Structural Biochemistry of a Bacterial Checkpoint Protein Reveals Diadenylate Cyclase Activity Regulated by DNA Recombination Intermediates. <i>Mol. Cell.</i> 30:167-178.
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<p>Reference:</p>
 
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1. <a href="http://www.jbc.org/content/288/5/3085" style="color:#303030" >Zhang <i>et al.</i> (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 18:02, 29 September 2013





The beast and its Achilles heel:

 A novel target to fight multi-resistant pathogenic bacteria



Array Team

By using the microarray technique we would like to analyze how many genes are regulated by ci-di-AMP in the Gram-positive model bacterium Bacillus subtilis. We are not only interested in the impact of c-di-AMP on the transcriptome, we also hope to identify novel regulatory elements (i.e. riboswitches) that bind to the signaling molecule. In our experiments we use three different B. subtilis strains. The wild-type strain contains all three diadenylatecyclases (DisA, CdaA, and CdaS). The second strain is a disA mutant strain lacking DisA, which was shown to be involved in DNA metabolism. This strain should produce less c-di-AMP than the isogenic parent strain. The third strain synthesizes a hyperactive CdaS mutant variant, which produces a lot of c-di-AMP. In our microarray experiments we will compare the transcriptomes of each mutant strain with that of the wild-type strain. Moreover, we would like to create a triple knock-out mutant (ΔdisA, ΔcdaA, ΔcdaS) lacking all diadenylate cyclases. We try to construct the triple knock-out mutant by feeding with exogenous c-di-AMP.

Reference:

1. Witte et al. (2008) Structural Biochemistry of a Bacterial Checkpoint Protein Reveals Diadenylate Cyclase Activity Regulated by DNA Recombination Intermediates. Mol. Cell. 30:167-178.


 

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