Team:UTK-Knoxville/HemAT

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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overall Project</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overall Project</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/methods"><span><span>Material &amp; Methods</span></span></a>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/methods"><span><span>Material &amp; Methods</span></span></a>
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</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION1</span></span></a></li>
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</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/partsorigin"><span><span>Parts Origin</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION2</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/construct"><span><span>Construct</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION3</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/Aer"><span>Aer</span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/Aer"><span>Aer</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/HemAT"><span>HemAT</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>HemAT</span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/RcoM"><span>RcoM</span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>RcoM</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/NarX"><span>NarX</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>TodS</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>LasR</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/EnvZ"><span>(EnvZ)</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>(EnvZ)</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/NtrC"><span>(NtrC)</span></a></li>
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                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>(NtrC)</span></a></li>
                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/primers"><span>Primers</span></a></li>
                     <li><a href="https://2013.igem.org/Team:UTK-Knoxville/primers"><span>Primers</span></a></li>
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  <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Characterization</span></span></a></li>
  <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Characterization</span></span></a></li>
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<ul class="DropDownMenu" id="MB1-DDM3">
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overview</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overview</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION1</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Background</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION2</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Application</span></span></a></li>
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<a href="https://2013.igem.org/Team:UTK-Knoxville/notebook" style="color: white">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]-->
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/notebook"><span><span>Daily Entries</span></span></a>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/notebook"><span>SECTION2</span></a></li>
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    </li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/methods"><span><span>Material &amp; Methods</span></span></a>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>SECTION3</span></a></li>
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  </li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Team Meetings</span></span></a>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>SECTION8</span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>SECTION9</span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>SECTION11</span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span>SECTION12</span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Visualization</span></span></a>
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</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Team Meetings</span></span></a>
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</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Seminar on DETAILS</span></span></a>
</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Seminar on DETAILS</span></span></a>
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<a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon" style="color: white">oUTreach<!--[if gt IE 6]><!--></a><!--<![endif]-->
<a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon" style="color: white">oUTreach<!--[if gt IE 6]><!--></a><!--<![endif]-->
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                              <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Project Overview</span></span></a></li>
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      <li><a href="https://2013.igem.org/Team:UTK-Knoxville/EastmanHITES"><span><span>Eastman HITES</span></span></a></li>
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      <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Section1</span></span></a></li>
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<a href="https://2013.igem.org/Team:UTK-Knoxville/sponsors" style="color: white">Sponsors</a>
<a href="https://2013.igem.org/Team:UTK-Knoxville/sponsors" style="color: white">Sponsors</a>
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<p>&nbsp;</p>
<p>&nbsp;</p>
<h3><span class="mw-headline" id="HemATDETAILS">HemAT</span></h3>
<h3><span class="mw-headline" id="HemATDETAILS">HemAT</span></h3>
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<p><img src="https://static.igem.org/mediawiki/2013/thumb/f/fd/HemAT_sensing_domain.png/647px-HemAT_sensing_domain.png" alt="HemAT" width="647" height="599"></p>
<p>Oxygen  gas, sensor FixL or HemAT (also need HemC).&nbsp;<a href="http://scop.berkeley.edu/sunid=111118">list of Direct Oxygen sensors </a>. <strong>FixL,</strong> was fused with <a href="http://www.sciencedirect.com/science/article/pii/S1570963902005551">5 different kinases</a> :  
<p>Oxygen  gas, sensor FixL or HemAT (also need HemC).&nbsp;<a href="http://scop.berkeley.edu/sunid=111118">list of Direct Oxygen sensors </a>. <strong>FixL,</strong> was fused with <a href="http://www.sciencedirect.com/science/article/pii/S1570963902005551">5 different kinases</a> :  
&ldquo;it was likely that the autophosphorylation activity of the  histidine kinase domain could not be regulated by the O2  association/dissociation in the sensor domain in the chimeric proteins&rdquo;  Intracellular &amp; redox sensor.&nbsp; <strong>O2 sensor, would be interesting.&nbsp; &amp; may also be able to sense&nbsp; CO in the absence of oxygen&nbsp; </strong>It seems like FixL is an O2 sensor.&nbsp; </p>
&ldquo;it was likely that the autophosphorylation activity of the  histidine kinase domain could not be regulated by the O2  association/dissociation in the sensor domain in the chimeric proteins&rdquo;  Intracellular &amp; redox sensor.&nbsp; <strong>O2 sensor, would be interesting.&nbsp; &amp; may also be able to sense&nbsp; CO in the absence of oxygen&nbsp; </strong>It seems like FixL is an O2 sensor.&nbsp; </p>
<p><a href="http://www.sciencedirect.com/science/article/pii/S0969212603001692">Sensing  domain is from 1-186</a>, its cytoplasmic, so it might not work on EnvZ.&nbsp; It also does not HAMP domain so we do not  think it would work on a <a href="http://aktuell.ruhr-uni-bochum.de/pm2012/pm00213.html.en">transmembrane kinase domain.</a></p>
<p><a href="http://www.sciencedirect.com/science/article/pii/S0969212603001692">Sensing  domain is from 1-186</a>, its cytoplasmic, so it might not work on EnvZ.&nbsp; It also does not HAMP domain so we do not  think it would work on a <a href="http://aktuell.ruhr-uni-bochum.de/pm2012/pm00213.html.en">transmembrane kinase domain.</a></p>
<p><a href="http://www.sciencedirect.com/science/article/pii/S1570963902005551">Chimeric  sensory kinases</a> containing O2 sensor domain of FixL and histidine kinase domain  from thermophile.</p>
<p><a href="http://www.sciencedirect.com/science/article/pii/S1570963902005551">Chimeric  sensory kinases</a> containing O2 sensor domain of FixL and histidine kinase domain  from thermophile.</p>
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<p>Oxygen&#8208;Sensing  Histidine&#8208;Protein Kinases:<a href="http://www.sciencedirect.com/science/article/pii/S0076687907370109"> Assays of Ligand Binding and Turnover of Response‐Regulator Substrates</a>.</p>
+
<p>Oxygen&#8208;Sensing  Histidine&#8208;Protein Kinases:<a href="http://www.sciencedirect.com/science/article/pii/S0076687907370109"> Assays of Ligand Binding and Turnover of Response-Regulator Substrates</a>.</p>
<p>Heme-based  oxygen sensor protein FixL:<a href="http://www.sciencedirect.com/science/article/pii/S0531513102002364"> its structure and function</a>.</p>
<p>Heme-based  oxygen sensor protein FixL:<a href="http://www.sciencedirect.com/science/article/pii/S0531513102002364"> its structure and function</a>.</p>
<p><a href="http://www.uniprot.org/uniprot/P76129#section_seq">Direct oxygen</a><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1305375/"> sensor protein</a>.</p>
<p><a href="http://www.uniprot.org/uniprot/P76129#section_seq">Direct oxygen</a><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1305375/"> sensor protein</a>.</p>
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<p><s>This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. </s></p>
<p><s>This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. </s></p>
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<h4>Resources:</h4>
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<h4 id="article-title-1" itemprop="headline"><a href="http://www.pnas.org/content/98/16/9353.full">Globin-coupled sensors: A class of heme-containing sensors in Archaea and Bacteria</a></h4>
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<p><a href="http://www.google.com/patents/US7129329">Heme proteins hemAT-Hs and hemAT-Bs and their use in medicine and microsensors</a></p>
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<h4 id="article-title-2" itemprop="headline"><a href="http://www.jbc.org/content/277/16/13528.full">Resonance Raman and Ligand Binding Studies of the Oxygen-sensing Signal Transducer Protein HemAT from&nbsp;<em>Bacillus subtilis</em></a><em></em></h4>
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<p itemprop="headline"><a href="http://www.hawaii.edu/microbiology/Alam/research/index.html">Globin-Coupled Sensors and Protoglobins</a></p>
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<p itemprop="headline">&nbsp;</p>
<p>&nbsp;</p>
<p>&nbsp;</p>
<p><br>
<p><br>

Latest revision as of 11:36, 5 August 2013

Team:UK-Knoxville - 2013.igem.org

HemAT

Aer plasmid KanR T7 OmpR Aer EnvZ pBR322

 

HemAT

HemAT

Oxygen gas, sensor FixL or HemAT (also need HemC). list of Direct Oxygen sensors . FixL, was fused with 5 different kinases : “it was likely that the autophosphorylation activity of the histidine kinase domain could not be regulated by the O2 association/dissociation in the sensor domain in the chimeric proteins” Intracellular & redox sensor.  O2 sensor, would be interesting.  & may also be able to sense  CO in the absence of oxygen  It seems like FixL is an O2 sensor. 

Sensing domain is from 1-186, its cytoplasmic, so it might not work on EnvZ.  It also does not HAMP domain so we do not think it would work on a transmembrane kinase domain.

Chimeric sensory kinases containing O2 sensor domain of FixL and histidine kinase domain from thermophile.

Oxygen‐Sensing Histidine‐Protein Kinases: Assays of Ligand Binding and Turnover of Response-Regulator Substrates.

Heme-based oxygen sensor protein FixL: its structure and function.

Direct oxygen sensor protein.

Amino Acid Sequence:
MLFKKDRKQETAYFSDSNGQQKNRIQLTNKHADVKKQLKMVRLGDAELYVLEQLQPLIQENIVNIVDAFYKNLDHESSL

MDIINDHSSVDRLKQTLKRHIQEMFAGVIDDEFIEKRNRIASIHLRIGLLPKWYMGAFQELLLSMIDIYEASITNQQELLKA

IKATTKILNLEQQLVLE

 

KanR

KanR provides kanamycin resistance to colonies with successful transformations.

 

pBR322

The pBR322 ori is a low copy origin of replication derived from

 

 

NtrB

Details for NtrB coming soon.Details for NtrB coming soon.Details for NtrB coming soon.

 

NtrC

Details for NtrC coming soon.Details for NtrC coming soon.Details for NtrC coming soon.

 

T7 Promoter

The T7 promoter is constitutively expressed by the phage T7 RNA polymerase. The polymerase is orthogonal to the wildtype E. coli promoters.

 

This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.

Gel samples

Gel Sample

This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.

 

Flow sample

FLowsample

 

This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go. This is where the deatils of Aer will go.

Resources:

Globin-coupled sensors: A class of heme-containing sensors in Archaea and Bacteria

Heme proteins hemAT-Hs and hemAT-Bs and their use in medicine and microsensors

Resonance Raman and Ligand Binding Studies of the Oxygen-sensing Signal Transducer Protein HemAT from Bacillus subtilis

Globin-Coupled Sensors and Protoglobins

 

 


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