Team:Gdansk-UG

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                 <td width="116" align="center" valign="middle"><p><a href="https://2013.igem.org/Team:Gdansk-UG/Parts">Parts Submitted</a></p>
                 <td width="116" align="center" valign="middle"><p><a href="https://2013.igem.org/Team:Gdansk-UG/Parts">Parts Submitted</a></p>
                   <a href="https://2013.igem.org/Team:Gdansk-UG/Parts">                  to the Registry</a></td>
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                 <td width="86" align="center" valign="middle"><a href="https://2013.igem.org/Team:Gdansk-UG/Modeling">Modeling</a></td>
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                 <td width="86" align="center" valign="middle"><a href="https://2013.igem.org/Team:Gdansk-UG/Modeling">Human practice</a></td>
                 <td width="99" align="center" valign="middle"><a href="https://2013.igem.org/Team:Gdansk-UG/Notebook">Notebook</a></td>
                 <td width="99" align="center" valign="middle"><a href="https://2013.igem.org/Team:Gdansk-UG/Notebook">Notebook</a></td>
                 <td width="68" align="center" valign="middle"><a href="https://2013.igem.org/Team:Gdansk-UG/Safety">Safety</a></td>
                 <td width="68" align="center" valign="middle"><a href="https://2013.igem.org/Team:Gdansk-UG/Safety">Safety</a></td>
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                         <td align="left" valign="top"><p>We are a group of students from Poland, Gda&#324;sk and we are better at biotechnology than at html.</p></td>
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                         <td align="left" valign="top"><p>We are a group of students from Poland, Gda&#324;sk and we present you our project: Metoli!</p></td>
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                             <p>We aim to construct bacteria able to detect  methanol in ethanol solutions. Our project focuses on using a methanol-dependent  promoter and a gene regulated by it &ndash; methanol dehydrogenase. We would like to  introduce bacteria, which &ndash;in presence of methanol &ndash; would produce a dye, for  instance, GFP. <br />
                             <p>We aim to construct bacteria able to detect  methanol in ethanol solutions. Our project focuses on using a methanol-dependent  promoter and a gene regulated by it &ndash; methanol dehydrogenase. We would like to  introduce bacteria, which &ndash;in presence of methanol &ndash; would produce a dye, for  instance, GFP. <br />
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                               The main problem is to find a promoter that  is insensible to ethanol. Unfortunately, the most suitable organism &ndash; <em>Picha pastoris</em> &ndash; has methanol-dependent  promoters which are blocked in the presence of ethanol. Therefore, we have  found a bacterium which is an obligate methylotroph &ndash; <em>Methylobacterium organophilum.</em> In theory, its methanol-dependent promoter should react only to methanol.<br />
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                               We would like to present two different  approaches to our project. Depending on initial results, we will take different  measures to reaching our goal. Being restricted by the possibility that the  promoter may not act as we would like it to, we thought of another route.<img width="611" height="492" src="https://static.igem.org/mediawiki/2013/b/b1/Wykres.jpg" /></p>
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                            <p>The main problem is to find a promoter that  is insensible to ethanol. Unfortunately, the most suitable organism &ndash; <em>Picha pastoris</em> &ndash; has methanol-dependent  promoters which are blocked in the presence of ethanol. Therefore, we have  found a bacterium which is an obligate methylotroph &ndash; <em>Methylobacillus flagellatus</em>. It is unable to utilize 2-carbon  compounds due to the absence of enzymes present in TCA cycle. Based on that  fact we speculate that the inhibition of the promoter will not take place.<br />
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                            <p>The first method, based on  methanol-dependent promoter is very clear and easy to achieve. Provided that  this promoter is insensible to ethanol and that it works well in our final,  transformed organism, we only have to measure the efficiency of dye production  in different concentrations of alcohols. <br />
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                               We would like to present two different  approaches to our project. Depending on initial results, we will take different  measures to reaching our goal. Being restricted by the possibility that the  promoter may not act as we would like it to, we thought of another route. </p>
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                               The other approach focuses on the reaction  catalyzed by methanol dehydrogenase. This enzyme, in the presence of NAD+,  is able to alter methanol into formaldehyde. While detecting the methanol with simple  measures is very hard, performing a colorimetric reaction with formaldehyde seems  to be quite possible. <br />
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The bacterium, which we want to use as a host is <em> E. coli </em> and if it works, we will try to perform the transformation on <em> Zymomonas mobilis</em>, which ethanol resistance is much hiigher. <em> Z. mobilis </em> transformation is much harder, so firstly we will focus on more known organism, <em> E.coli </em>.
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                              The first method, based on  methanol-dependent promoter is very clear and easy to achieve. Provided that  this promoter is insensible to ethanol and that it works well in our final,  transformed organism, we only have to measure the efficiency of dye production  in different concentrations of alcohols. <br />
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                               The other approach focuses on the reaction  catalyzed by methanol dehydrogenase. This enzyme, in the presence of NAD+,  is able to alter methanol into formaldehyde. <br />
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                              While detecting the methanol with simple  measures is very hard, performing a colorimetric reaction with formaldehyde seems  to be quite possible. We are hoping that the enzyme shows higher affinity to  methanol than to ethanol, but even if it doesn&rsquo;t &ndash; this method would still be sufficiently  precise to detect low concentrations of one-carbon alcohol.<br />
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                            Taking into account that we may decide to follow the second route, we are also gathering information about detecting  formaldehyde in purely biological ways. We found a formaldehyde operon but, unfortunately, there is not enough information about it.&nbsp; We are hoping to find either an enzyme,  promoter or an operon that would enable us to insert into bacteria a sequence  resulting in producing colored substance during bacterial growth in methanol. </p></td>
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Latest revision as of 16:49, 3 October 2013

Gdansk UG

Home Team

Official

Team Profile

Project

Parts Submitted

to the Registry
Human practice Notebook Safety Attributions

Welcome to our Site

 

We are a group of students from Poland, Gdańsk and we present you our project: Metoli!

 

 

We aim to construct bacteria able to detect methanol in ethanol solutions. Our project focuses on using a methanol-dependent promoter and a gene regulated by it – methanol dehydrogenase. We would like to introduce bacteria, which –in presence of methanol – would produce a dye, for instance, GFP.
The main problem is to find a promoter that is insensible to ethanol. Unfortunately, the most suitable organism – Picha pastoris – has methanol-dependent promoters which are blocked in the presence of ethanol. Therefore, we have found a bacterium which is an obligate methylotroph – Methylobacterium organophilum. In theory, its methanol-dependent promoter should react only to methanol.
We would like to present two different approaches to our project. Depending on initial results, we will take different measures to reaching our goal. Being restricted by the possibility that the promoter may not act as we would like it to, we thought of another route.

The first method, based on methanol-dependent promoter is very clear and easy to achieve. Provided that this promoter is insensible to ethanol and that it works well in our final, transformed organism, we only have to measure the efficiency of dye production in different concentrations of alcohols.
The other approach focuses on the reaction catalyzed by methanol dehydrogenase. This enzyme, in the presence of NAD+, is able to alter methanol into formaldehyde. While detecting the methanol with simple measures is very hard, performing a colorimetric reaction with formaldehyde seems to be quite possible.

The bacterium, which we want to use as a host is E. coli and if it works, we will try to perform the transformation on Zymomonas mobilis, which ethanol resistance is much hiigher. Z. mobilis transformation is much harder, so firstly we will focus on more known organism, E.coli .