Team:Goettingen/Team/Array
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===Array Team=== | ===Array Team=== | ||
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- | <p> The genome of the Gram-positive model bacterium <i>Bacillus subtilis</i> contains the <i>disA, cdaA</i>, and </i>cdaS</i> genes encoding enzymatically active adenylate cyclases (Mehne <i>et al.</i>). By using the microarray technique we would like to analyze how many genes are regulated by c-di-AMP in <i>B. 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> | + | <p> The genome of the Gram-positive model bacterium <i>Bacillus subtilis</i> contains the <i>disA, cdaA</i>, and </i>cdaS</i> genes encoding enzymatically active adenylate cyclases (Mehne <i>et al.</i> 2013). By using the microarray technique we would like to analyze how many genes are regulated by c-di-AMP in <i>B. 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 (Witte <i>et al.</i> 2008). 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> | ||
- | 1. Mehne <i>et al.</i> (2013) | + | 1. Mehne <i>et al.</i> (2013) Cyclic di-AMP homeostasis in <i>Bacillus subtilis</i>: both lack and high level accumulation of the nucleotide are detrimental for cell growth. <i>J. Biol. Chem.</i> 288:2004-2017. |
2. 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. | 2. 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. |
Revision as of 18:22, 29 September 2013