Team:TMU-Tokyo/Project device1

From 2012.igem.org

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This device is composed of <i>frmR</i> gene, its promoter and GFP gene.</p>
This device is composed of <i>frmR</i> gene, its promoter and GFP gene.</p>
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<center><Img Src="https://static.igem.org/mediawiki/2012/3/3a/Device1.png"></center><Br>
<center><Img Src="https://static.igem.org/mediawiki/2012/3/3a/Device1.png"></center><Br>
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<B>■Details of <i>frmR</i> effect</B>
<B>■Details of <i>frmR</i> effect</B>
<Br>Christopher D. Herring and Frederick R. Blattner. 2004. "Global Transcriptional Effects of a Suppressor tRNA and the Inactivation of the Regulator <i>frmR</i>" [<A Href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC522192/">PubMed</A>]<Br>
<Br>Christopher D. Herring and Frederick R. Blattner. 2004. "Global Transcriptional Effects of a Suppressor tRNA and the Inactivation of the Regulator <i>frmR</i>" [<A Href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC522192/">PubMed</A>]<Br>
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fig1. a) When formaldehyde doesn't exist, <i>frmR</i> represses upstream promoter. In this state, transcription is stopped and GFP isn't produced. b) When formaldehyde exists, it makes <i>frmR</i> lose activity. Then promoter is released from <i>frmR</i>'s repress and begins to produce GFP.<Br>
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<b>■fig1.<Br>
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a)</b> When formaldehyde doesn't exist, <i>frmR</i> represses upstream promoter. In this state, transcription is stopped and GFP isn't produced.<Br>
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<b>b)</b> When formaldehyde exists, it makes <i>frmR</i> lose activity. Then promoter is released from <i>frmR</i>'s repress and begins to produce GFP.<Br>
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Repressing ability of <i>frmR</i> was not shown molecularly but quantitatively. The downstream of frmR is inhibited by frmR and not by frmR added 7 amino acids.<Br>
Repressing ability of <i>frmR</i> was not shown molecularly but quantitatively. The downstream of frmR is inhibited by frmR and not by frmR added 7 amino acids.<Br>
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<center><p class="description"><Img Src="https://static.igem.org/mediawiki/2012/6/6c/FrmR.jpg" Width="53%" Height="53%">
<center><p class="description"><Img Src="https://static.igem.org/mediawiki/2012/6/6c/FrmR.jpg" Width="53%" Height="53%">
<Img Src="https://static.igem.org/mediawiki/2012/f/ff/FrmR2.jpg" Width="44% Height="44%" ><Br>
<Img Src="https://static.igem.org/mediawiki/2012/f/ff/FrmR2.jpg" Width="44% Height="44%" ><Br>
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G Eggertsson and D Söll. 1988. "Transfer ribonucleic acid-mediated suppression of termination codons in <i>Escherichia coli</i>."[<A Href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC373150/">PMC</A>]<Br>
G Eggertsson and D Söll. 1988. "Transfer ribonucleic acid-mediated suppression of termination codons in <i>Escherichia coli</i>."[<A Href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC373150/">PMC</A>]<Br>
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<B>■Expriment of <i>frmR</i> quantitative repressing detection</B><Br>
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<B>■Function of <i>frmR</i> quantitative Analysis</B><Br>
Christopher D. Herring and Frederick R. Blattner. 2004. "Global Transcriptional Effects of a Suppressor tRNA and the Inactivation of the Regulator <i>frmR</i>" [<A Href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC522192/">PubMed</A>]<Br>
Christopher D. Herring and Frederick R. Blattner. 2004. "Global Transcriptional Effects of a Suppressor tRNA and the Inactivation of the Regulator <i>frmR</i>" [<A Href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC522192/">PubMed</A>]<Br>
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<B>■fig2.(left)<Br>
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<B>■fig3.(right)<Br>
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Revision as of 04:21, 15 September 2012

 


Device1;

Sensing formaldehyde and then producing GFP



 
 

In frmRAB operon, which origin is chromosome of Escherichia coli, frmAB is normally repressed by frmR. But, when formaldehyde exists, frmR loses its inhibitory and frmAB is expressed.
This device is composed of frmR gene, its promoter and GFP gene.




By using this device, we can visualize formaldehyde with GFP. This visualization system enable to make us know how much of the formaldehyde concentration there is.


■Details of frmR effect
Christopher D. Herring and Frederick R. Blattner. 2004. "Global Transcriptional Effects of a Suppressor tRNA and the Inactivation of the Regulator frmR" [PubMed]

■fig1.
a)
When formaldehyde doesn't exist, frmR represses upstream promoter. In this state, transcription is stopped and GFP isn't produced.
b) When formaldehyde exists, it makes frmR lose activity. Then promoter is released from frmR's repress and begins to produce GFP.

 
 

Details of frmR repression



 
 

The sequence of frmR contains amber suppressor codon (UAG) at its 3' end. Amber suppressor codon recognizes its specialized tRNA. But the tRNA sometimes alters and gets capacity to carry amino acid. If amino acid comes, transcription is continued to more 7 amino acids and cording protein is degenerated. Wherein, frmR loses inhibitory.

Repressing ability of frmR was not shown molecularly but quantitatively. The downstream of frmR is inhibited by frmR and not by frmR added 7 amino acids.







Origin of frmR is guessed same as CsoR, copper-sensitive operon repressor. Both sequence is well preserved. It has been shown that CsoR represses expression of genes located in downstream. So we guess it is same as frmR.


■Details of amber suppressor
G Eggertsson and D Söll. 1988. "Transfer ribonucleic acid-mediated suppression of termination codons in Escherichia coli."[PMC]

■Function of frmR quantitative Analysis
Christopher D. Herring and Frederick R. Blattner. 2004. "Global Transcriptional Effects of a Suppressor tRNA and the Inactivation of the Regulator frmR" [PubMed]

■fig2.(left)
■fig3.(right)