Team:Wisconsin-Madison/biosensor
From 2011.igem.org
(7 intermediate revisions not shown) | |||
Line 157: | Line 157: | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison">Main</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison">Main</a> | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison/whatisigem">What is iGEM?</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/whatisigem">What is iGEM?</a> | ||
+ | <a href="https://2011.igem.org/Team:Wisconsin-Madison/ca">Contributions & Attributions</a> | ||
</div> | </div> | ||
</li> | </li> | ||
Line 166: | Line 167: | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">Directed Evolution</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">Directed Evolution</a> | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison/bmc">Microcompartment</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/bmc">Microcompartment</a> | ||
+ | <a href="https://2011.igem.org/Team:Wisconsin-Madison/parts">Parts</a> | ||
</div> | </div> | ||
</li> | </li> | ||
Line 179: | Line 181: | ||
<div id="m4" onmouseover="mcancelclosetime()" onmouseout="mclosetime()"> | <div id="m4" onmouseover="mcancelclosetime()" onmouseout="mclosetime()"> | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison/protocols">Protocols</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/protocols">Protocols</a> | ||
- | <a href="https://2011.igem.org/Team:Wisconsin-Madison/calender"> | + | <a href="https://2011.igem.org/Team:Wisconsin-Madison/calender">Calendar</a> |
<a href="https://2011.igem.org/Team:Wisconsin-Madison/references">References</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/references">References</a> | ||
</div> | </div> | ||
Line 188: | Line 190: | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison/reuposterSession">REU Poster Session</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/reuposterSession">REU Poster Session</a> | ||
<a href="https://2011.igem.org/Team:Wisconsin-Madison/socialmedia">Social Media</a> | <a href="https://2011.igem.org/Team:Wisconsin-Madison/socialmedia">Social Media</a> | ||
- | |||
</div> | </div> | ||
</li> | </li> | ||
<li><a href="https://2011.igem.org/Team:Wisconsin-Madison/safety" onmouseover="mopen('m6')" onmouseout="mclosetime()">Safety</a> | <li><a href="https://2011.igem.org/Team:Wisconsin-Madison/safety" onmouseover="mopen('m6')" onmouseout="mclosetime()">Safety</a> | ||
+ | <div id="m6" onmouseover="mcancelclosetime()" onmouseout="mclosetime()"> | ||
+ | <a href="https://2011.igem.org/Team:Wisconsin-Madison/humanpractice">Human Practice</a> | ||
+ | </div> | ||
</li> | </li> | ||
</ul> | </ul> | ||
</center> | </center> | ||
- | + | ||
<center> | <center> | ||
<img src="https://static.igem.org/mediawiki/2011/a/aa/Logo_v2.2.jpg"; style="position:absolute; left:-10px; top:-10px"/> | <img src="https://static.igem.org/mediawiki/2011/a/aa/Logo_v2.2.jpg"; style="position:absolute; left:-10px; top:-10px"/> | ||
</center> | </center> | ||
+ | |||
Line 219: | Line 224: | ||
Both the ethanol (EtOH) and n-alkane sensors rely on an arabinose inducible two plasmid system for either EtOH or n-alkane detection. The first plasmid contains genes for producing a protein controlled the pBAD promoter, which turns on transcription of the <a href="https://2011.igem.org/Team:Wisconsin-Madison/genes">genes</a> in the presence of arabinose. The second plasmid has the EtOH or n-alkane promoter which turns on the transcription of our RFP. The production of the RFP is then measured with a plate reader. | Both the ethanol (EtOH) and n-alkane sensors rely on an arabinose inducible two plasmid system for either EtOH or n-alkane detection. The first plasmid contains genes for producing a protein controlled the pBAD promoter, which turns on transcription of the <a href="https://2011.igem.org/Team:Wisconsin-Madison/genes">genes</a> in the presence of arabinose. The second plasmid has the EtOH or n-alkane promoter which turns on the transcription of our RFP. The production of the RFP is then measured with a plate reader. | ||
- | <p>< | + | <p><center> |
- | <img src="https://static.igem.org/mediawiki/2011/4/49/2_prasmid.jpg" width = " | + | <img src="https://static.igem.org/mediawiki/2011/4/49/2_prasmid.jpg" width = "550"/> |
- | <p> | + | <p></center> |
In the <a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">directed evolution</a> construct, this two plasmid system is all put on a single plasmid, which the first plasmid’s parts being put on the reverse strand (5’ to 3’) of the DNA. | In the <a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">directed evolution</a> construct, this two plasmid system is all put on a single plasmid, which the first plasmid’s parts being put on the reverse strand (5’ to 3’) of the DNA. | ||
+ | <p> | ||
+ | <center><img src="https://static.igem.org/mediawiki/2011/8/89/Prasmid_2.jpg" width = "550"/></center> | ||
+ | <p> | ||
+ | In our case, it is entirely possible to use a constitutive promoter in place of the current arabinose inducible promoter, as the first part of the system does not rely on the presence of either EtOH or an n-alkane to function. However, by controlling the transcription on the first plasmid via the presence or absence of arabinose, we can a) determine the leakiness of the second plasmid, which relies on the transcription of the first genes, and b) it gives us better control over the activation of our sensor. The pBAD promoter’s activity in the presence of arabinose has a sigmoidal shape; that is, below the saturation concentration of arabinose, the promoter is ‘off’ and above the saturation concentration it is ‘on’ (i.e. there is no gradient dependence on arabinose) It is worth noting, however, that the pBAD promoter is notoriously leaky, and that is may be more effective to suppress its activity with glucose, instead of increasing its activity with arabinose. | ||
<p><br> | <p><br> | ||
- | Learn more about <a href="https://2011.igem.org/Team:Wisconsin-Madison/biofuels">biofuels</a>, <a href="https://2011.igem.org/Team:Wisconsin-Madison/genes">genes</a>. | + | Learn more about <a href="https://2011.igem.org/Team:Wisconsin-Madison/biofuels">biofuels</a>, <a href="https://2011.igem.org/Team:Wisconsin-Madison/genes">genes</a>, <a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">directed evolution</a>. |
Latest revision as of 22:59, 28 September 2011
Project >>
Overview,
Ethanol Sensor,
Alkane Sensor,
Microcompartment
Biosensors
Both the ethanol (EtOH) and n-alkane sensors rely on an arabinose inducible two plasmid system for either EtOH or n-alkane detection. The first plasmid contains genes for producing a protein controlled the pBAD promoter, which turns on transcription of the genes in the presence of arabinose. The second plasmid has the EtOH or n-alkane promoter which turns on the transcription of our RFP. The production of the RFP is then measured with a plate reader.
In our case, it is entirely possible to use a constitutive promoter in place of the current arabinose inducible promoter, as the first part of the system does not rely on the presence of either EtOH or an n-alkane to function. However, by controlling the transcription on the first plasmid via the presence or absence of arabinose, we can a) determine the leakiness of the second plasmid, which relies on the transcription of the first genes, and b) it gives us better control over the activation of our sensor. The pBAD promoter’s activity in the presence of arabinose has a sigmoidal shape; that is, below the saturation concentration of arabinose, the promoter is ‘off’ and above the saturation concentration it is ‘on’ (i.e. there is no gradient dependence on arabinose) It is worth noting, however, that the pBAD promoter is notoriously leaky, and that is may be more effective to suppress its activity with glucose, instead of increasing its activity with arabinose.
|