Team:LCG-UNAM-Mexico/Project

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               The aim of this project is to make bacteria sing,  and it will be done by modifying the extracellular  medium's resistivity through the modulation of the RcnA (<em>E. coli'</em>s natural efflux pump) which will in turn change the concentration of nickel  outside the cell. By doing this we expect to gain insight into a  fundamental aspect of ecological dynamics which is currently not well  understood. On the other hand, we expect to show that measuring changes  in resistivity is an effective way to determine the activity of the  efflux pump, and that this can become an efficient indicator of real  time transcription for <em>in vivo</em> experiments.   </p>
               The aim of this project is to make bacteria sing,  and it will be done by modifying the extracellular  medium's resistivity through the modulation of the RcnA (<em>E. coli'</em>s natural efflux pump) which will in turn change the concentration of nickel  outside the cell. By doing this we expect to gain insight into a  fundamental aspect of ecological dynamics which is currently not well  understood. On the other hand, we expect to show that measuring changes  in resistivity is an effective way to determine the activity of the  efflux pump, and that this can become an efficient indicator of real  time transcription for <em>in vivo</em> experiments.   </p>
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             <p align="justify"> <span class="bodyText">To achieve this, we designed a system whe</span>re <em>rcnA</em> is under  the regulation of the lambda repressor CI which itself is under the regulation  of <em>Vibrio fisheri</em>’s quorum sensing core components: LuxR and AHL. The  former is constitutively produced and the latter is our input signal.  Further regulation of the system is achieved by constitutively  synthesizing AiiA, which degrades AHL, and by RcnR, RcnA's natural repressor which is inactivated in the presence of nickel.  All  the protein components of the system are encoded in <a href="https://2008.igem.org/Team:LCG-UNAM-Mexico/Parts">two devices</a>:  the first one contains the efflux pump, and the second one the  regulatory cascade. The reason for this partitioning is that in this  way RcnA can be integrated with any desired regulatory upstream signal.   </p>
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             <p align="justify" class="bodyText"> To achieve this, we designed a system where <em>rcnA</em> is under  the regulation of the lambda repressor CI which itself is under the regulation  of <em>Vibrio fisheri</em>’s quorum sensing core components: LuxR and AHL. The  former is constitutively produced and the latter is our input signal.  Further regulation of the system is achieved by constitutively  synthesizing AiiA, which degrades AHL, and by RcnR, RcnA's natural repressor which is inactivated in the presence of nickel.  All  the protein components of the system are encoded in <a href="https://2008.igem.org/Team:LCG-UNAM-Mexico/Parts">two devices</a>:  the first one contains the efflux pump, and the second one the  regulatory cascade. The reason for this partitioning is that in this  way RcnA can be integrated with any desired regulatory upstream signal.   </p>
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             <p align="justify"> Once the system is built and implemented on <em>E.  coli</em>, we will measure the medium's resistivity on a real time basis  through a set of copper electrodes connected to a multimeter. The  monitoring device will be connected to a computer which will filter the  noise in the signal and return a sound depending on the resistivity. <a href="http://en.wikiquote.org/wiki/Frankenstein_(1931_film)">They sing! They sing! Now we know what it’s like to be God!</a></p>
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             <p align="justify" class="bodyText"> Once the system is built and implemented on <em>E.  coli</em>, we will measure the medium's resistivity on a real time basis  through a set of copper electrodes connected to a multimeter. The  monitoring device will be connected to a computer which will filter the  noise in the signal and return a sound depending on the resistivity. <a href="http://en.wikiquote.org/wiki/Frankenstein_(1931_film)">They sing! They sing! Now we know what it’s like to be God!</a></p>
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            <p align="justify">&nbsp;</p>
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Latest revision as of 22:18, 28 October 2008

LCG-UNAM-Mexico:Modeling

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♪ Singing bacteria! ♫


Controlling E. coli's nickel efflux pump

I bet you're thinking we're insane... so why don't you take a look at out project? Below you can watch a Flash movie summarizing the whole idea:


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Project description


The aim of this project is to make bacteria sing, and it will be done by modifying the extracellular medium's resistivity through the modulation of the RcnA (E. coli's natural efflux pump) which will in turn change the concentration of nickel outside the cell. By doing this we expect to gain insight into a fundamental aspect of ecological dynamics which is currently not well understood. On the other hand, we expect to show that measuring changes in resistivity is an effective way to determine the activity of the efflux pump, and that this can become an efficient indicator of real time transcription for in vivo experiments.  

To achieve this, we designed a system where rcnA is under the regulation of the lambda repressor CI which itself is under the regulation of Vibrio fisheri’s quorum sensing core components: LuxR and AHL. The former is constitutively produced and the latter is our input signal. Further regulation of the system is achieved by constitutively synthesizing AiiA, which degrades AHL, and by RcnR, RcnA's natural repressor which is inactivated in the presence of nickel.  All the protein components of the system are encoded in two devices: the first one contains the efflux pump, and the second one the regulatory cascade. The reason for this partitioning is that in this way RcnA can be integrated with any desired regulatory upstream signal.  

Once the system is built and implemented on E. coli, we will measure the medium's resistivity on a real time basis through a set of copper electrodes connected to a multimeter. The monitoring device will be connected to a computer which will filter the noise in the signal and return a sound depending on the resistivity. They sing! They sing! Now we know what it’s like to be God!

 

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