Difference between revisions of "Team:UMaryland/Part Collection"

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{{UMaryland}}
 
<html>
 
  
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</div>
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</div>
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<html lang="en">
  
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<head>
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    <script>
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        $('body').css('opacity', '0');
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    <script src="https://static.igem.org/mediawiki/2016/9/95/T--UMaryland--html5shiv.txt"></script>
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    <title>Modeling</title>
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<body>
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            font-family: 'Exo';
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        @font-face {
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/* Slider */ .slick-slider { position: relative; display: block; box-sizing: border-box; -webkit-user-select: none; -moz-user-select: none; -ms-user-select: none; user-select: none; -webkit-touch-callout: none; -khtml-user-select: none; -ms-touch-action: pan-y; touch-action: pan-y; -webkit-tap-highlight-color: transparent; } .slick-list { position: relative; display: block; overflow: hidden; margin: 0; padding: 0; } .slick-list:focus { outline: none; } .slick-list.dragging { cursor: pointer; cursor: hand; } .slick-slider .slick-track, .slick-slider .slick-list { -webkit-transform: translate3d(0, 0, 0); -moz-transform: translate3d(0, 0, 0); -ms-transform: translate3d(0, 0, 0); -o-transform: translate3d(0, 0, 0); transform: translate3d(0, 0, 0); } .slick-track { position: relative; top: 0; left: 0; display: block; } .slick-track:before, .slick-track:after { display: table; content: ''; } .slick-track:after { clear: both; } .slick-loading .slick-track { visibility: hidden; } .slick-slide { display: none; float: left; height: 100%; min-height: 1px; } [dir='rtl'] .slick-slide { float: right; } .slick-slide img { display: block; } .slick-slide.slick-loading img { display: none; } .slick-slide.dragging img { pointer-events: none; } .slick-initialized .slick-slide { display: block; } .slick-loading .slick-slide { visibility: hidden; } .slick-vertical .slick-slide { display: block; height: auto; border: 1px solid transparent; } .slick-arrow.slick-hidden { display: none; } .slick-arrow { background-color: #B0B0B0;}
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<style>
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@charset "UTF-8";
  
<div class="column full_size judges-will-not-evaluate">
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/*!
<h3>★ ALERT! </h3>
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  * animate.css -http://daneden.me/animate
<p>This page is used by the judges to evaluate your team for the <a href="https://2016.igem.org/Judging/Awards#Special_Prizes">Part Collection special prize</a>. </p>
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* Version - 3.5.1
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* Licensed under the MIT license - http://opensource.org/licenses/MIT
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*
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* Copyright (c) 2016 Daniel Eden
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*/
  
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 +
</style>
 +
 +
<style>
 +
/* Permanent bottom bar */
 +
 +
#bot-nav {
 +
    background-color: #f9f9f9;   
 +
    border-top: 1px solid #f5f5f5;
 +
    position: absolute;
 +
    width: 100%;
 +
    height: 100px;
 +
    display: block;
 +
    color: black;
 +
    font-size: 12pt;
 +
    font-family: 'Lora', serif;
 +
    z-index: -1;
 +
}
  
 +
#social-media {
 +
width: 100%;
 +
margin:0 0 0 0;
 +
}
 +
.pull-right a {
 +
padding-right: 0px !important;
 +
}
  
<p> Delete this box in order to be evaluated for this medal. See more information at <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions"> Instructions for Pages for awards</a>.</p>
+
.copyright {
</div>
+
text-align: center;
 +
width: 272px;
 +
height: 30px;
 +
margin-top: -20px;
 +
margin-left: auto;
 +
margin-right: auto;
 +
}
  
 +
.pull-left {
 +
margin-top: -3px;
 +
width: 83px;
 +
margin-left: 30px;
 +
margin-right: auto;
 +
display: inline;
 +
height: 34px;
 +
}
  
 +
.pull-left p {
 +
margin: 0 0 0 0;
 +
padding: 0 0 0 0 !important;
 +
line-height: 0px !important;
 +
}
  
 +
.pull-right {
 +
width: 83px;
 +
margin-right: 30px;
 +
margin-left: auto;
 +
}
  
 +
#copyright {
 +
    text-align: center;
 +
left:auto;
 +
right: auto;
 +
width: 100%;
 +
padding: 0 0 0 0;
 +
margin-top: 0px;
 +
float:left;
 +
}
  
<div class="column full_size">
+
#container {
 +
    max-width: 1200px;
 +
    margin-left: auto;
 +
    margin-right: auto;
 +
margin-top: 40px;
 +
}
  
 +
#container img {
 +
    height: 20px;
 +
    width: 20px;
 +
    -webkit-filter: grayscale(100%);
 +
    filter: grayscale(100%);
 +
}
  
 +
#bot-nav img:hover {
 +
    -webkit-filter: grayscale(0%);
 +
    filter: grayscale(0%);
 +
}
  
 +
#container li {
 +
    display: inline;
 +
list-style:none;
 +
padding-right: 25px;
 +
}
  
 +
.pull-left {
 +
float:left;
 +
}
  
<p>Did your team make a lot of great parts? Is there a theme that ties all your parts together? Do you have more than 10 parts in this collection? Did you make a CRISPR collection, a MoClo collection, or a collection of awesome pigment parts? Describe your parts collection on this page, so the judges can evaluate you for the Best Part Collection award.</p>
+
#bot-nav a {
 +
    text-decoration: none;
 +
    color: black;
 +
}
  
<p>
+
#contact:hover {
While you should put all the characterization information for your parts on the Registry, you are encouraged to explain how all your parts form a collection on this page.  
+
    text-shadow: .1px .1px black;
</p>
+
}
 +
</style>
 +
    <style>
 +
        /* Navbar and body setup */
 +
        #bars_item {
 +
            display: none;
 +
        }
  
 +
        #sideMenu {
 +
            display: none;
 +
        }     
  
<div class="highlight">
+
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<h4>Note</h4>
+
                margin-top: 0px;
<p>This page should list all the parts in the collection your team made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page.</p>
+
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<nav>
 +
        <a href="https://2016.igem.org/Team:UMaryland">
 +
<img src="https://static.igem.org/mediawiki/2016/2/26/T--UMaryland--newlogo.jpeg" id="img-logo" width="128px">
 +
        </a>
 +
<ul>
 +
<a style="opacity: 0">Fix</a>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/projects">Projects</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/Part_Collection">Parts</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/Model">Modeling</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/outreach">Outreach</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/notebook">Notebook</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/about">About Us</a></li>
 +
</ul>
 +
</nav>
 +
<img src="https://static.igem.org/mediawiki/2016/e/e5/T--UMaryland--more.png" id="img-more">
 +
<!-- Nav bar when more button is clicked -->
 +
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 +
<a href="https://2016.igem.org/Team:UMaryland"><img src="https://static.igem.org/mediawiki/2016/2/26/T--UMaryland--newlogo.jpeg" id="img-logoResized"></a>
 +
<ul>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/projects">Projects</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/Part_Collection">Parts</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/Model">Modeling</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/outreach">Outreach</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/notebook">Notebook</a></li>
 +
<li><a href="https://2016.igem.org/Team:UMaryland/about">About Us</a></li>
 +
</ul>
 +
</div>
 +
<div data-parallax="scroll" data-image-src="https://static.igem.org/mediawiki/2016/6/69/T--UMaryland--parts.jpg" class="div-scrollPic" style="height: 650px;">
 +
<div class="titleText">
 +
<!-- where background image below titles show -->
 +
<h11>Parts Collection</h11></br>
 +
<h21>BioBrick Devices Submitted to the Registry</h21></br>
 +
<h31>Furthering collaboration and standardization of genetic parts</h31></br>
 +
</div>
 +
        </div>
 +
<div id="div-text"> <!-- Start of main text of page -->
 +
<!-- Heading text -->
 +
<div class="longText">
 +
<p>UMaryland iGEM submitted various basic and composite parts to the BioBrick Registry, which aims to increase standardization in synthetic biology by allowing genes to be added together easily. We synthesized the genes, put them inside standard BioBrick plasmids, and then characterized our parts.</p>
 +
<p>View:</p>
 +
<p>
 +
Basic Parts <input type="checkbox" class="filter" checked id="input-basic" />
 +
Composite Parts <input type="checkbox" class="filter" checked id="input-composite" />
 +
</p>
 +
</div>
 +
<style>
 +
input {
 +
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 +
 +
.longText strong {
 +
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 +
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 +
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 +
 +
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 +
margin-top: 30px;
 +
margin-bottom: 30px;
 +
}
 +
 +
 +
 +
.profiles {
 +
border: 2px solid #A8A8A8;
 +
width: 1000px;
 +
margin-left: 20px;
 +
margin-right: 20px;
 +
-webkit-border-radius: 5px;
 +
-moz-border-radius: 5px;
 +
border-radius: 5px;
 +
}
 +
 +
.profiles p {
 +
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 +
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 +
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 +
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 +
margin-top: 30px !important;
 +
margin-bottom: 0px !important;
 +
margin-right: 15px !important;
 +
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 +
 +
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 +
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 +
margin-top: 15px;
 +
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 +
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 +
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 +
text-decoration: underline;
 +
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 +
 +
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 +
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 +
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 +
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 +
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 +
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 +
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 +
 +
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 +
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 +
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 +
margin-top: 10px;
 +
margin-bottom: 15px;
 +
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 +
 +
#div-slideshow {
 +
margin-top: 40px;
 +
margin-bottom: 40px;
 +
margin-left: auto;
 +
margin-right: auto;
 +
width: 1000px;
 +
padding-bottom: 30px;
 +
overflow: normal;
 +
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 +
 +
</style>
 +
<div id="div-slideshow">
 +
<div class="profiles composite">
 +
<strong><a target="_blank" href="http://parts.igem.org/Part:BBa_K2032001">Fructose Pathway (BBa_K2032001)</strong></a>
 +
<img src="https://static.igem.org/mediawiki/2016/e/e7/T--UMaryland--BBa_K2032001_linear.jpg" class="linear" />
 +
<img src="https://static.igem.org/mediawiki/2016/1/14/T--UMaryland--BBa_K2032001_plasmid.jpg" class="plasmid" />
 +
<p>This composite biobrick part is a combination of the coding regions for three separate enzymes involved in the metabolization of methanol. Each coding region is preempted by a ribosome binding site in order to help counteract some of the translational issues associated with polycistronic mRNA. A lacI regulated promoter that allows for induction with IPTG was included in this construct to allow for selective gene expression along with the standard iGEM double terminator.</br></br>The three enzymes encoded by this part are methanol dehydrogenase 2 (MDH), 3-hexulose-6-phosphate synthase (HPS), and 6-Phospho-3-hexuloisomerase (PHI). These enzymes serve to function as a three step pathway in which methanol is metabolized. MDH converts methanol to formaldehyde, producing a molecule of NADH in the process. Formaldehyde is then combined with a molecule of D-ribulose-5-phosphate taken from the pentose phosphate pathway to form one molecule of D-arabino-3-hexulose-6-phosphate via the usage of HPS. PHI then converts D-arabino-3-hexulose-6-phosphate to D-fructose-6-phosphate which can then undergo glycolysis.</p>
 +
</div>
 +
<div class="profiles basic">
 +
<strong><a target="_blank" href="http://parts.igem.org/Part:BBa_K2032002">Formate Pathway (BBa_K2032002)</a></strong>
 +
<img src="https://static.igem.org/mediawiki/2016/9/9b/T--UMaryland--BBa_K2032002_linear.jpg" class="linear" style="height: 120px !important;"/>
 +
<img src="https://static.igem.org/mediawiki/2016/a/a6/T--UMaryland--BBa_K2032002.png" class="plasmid" />
 +
<p>The Formate Pathway is a three enzyme pathway that begins with methanol and NAD+ as substrates, and culminates in the production of NADH molecules and carbon dioxide. This pathway is used to detoxify alcohols in the cellular environment. The pathway consists of a series of oxidations: methanol oxidized to formaldehyde by Methanol Dehydrogenase 2 (MDH2); formaldehyde oxidized to formate by Formaldehyde Dehydrogenase (FALDH); and finally formate oxidized to carbon dioxide by Formate Dehydrogenase (FDH). Each of these catalyzed reactions results in lower energy products than reactants, so every reaction is coupled to the production of one NADH molecule, which contributes to energy for the cell and biomass production.</br></br> The first enzyme of the pathway, MDH2, has a low binding specificity and will oxidize many primary alcohols. The original use of this plasmid was to detoxify methanol as part of a larger pathway that consisted of eliminating methane gas from the atmosphere. The pathway has potential uses for detoxifying alcohols in the environment as well. As methanol becomes an increasingly popular liquid fuel source, one could imagine methanol spills in the future that require detoxification. This part contains one promoter that is IPTG inducible. The genes will be transcribed as a polycistronic mRNA strand. Each of the genes has a medium strength ribosome binding site. Modeling of this pathway has revealed that no toxic substrates should be produced when this pathway is expressed. The kinetics for each enzyme exist in such a way that there will be no buildup of formaldehyde of formic acid in the cell. Theoretically, this pathway should increase methanol resistance to cells that express it.</p>
 +
</p>
 +
</div>
 +
<div class="profiles basic">
 +
<strong><a target="_blank" href="http://parts.igem.org/Part:BBa_K2032003">Codon optimized MDH2 with Lac/pL promoter (BBa_K2032003)</strong></a>
 +
<img src="https://static.igem.org/mediawiki/2016/8/8c/T--UMaryland--BBa_K2032003_plasmid.jpg" class="plasmid" />
 +
<p>This is an intermediate used in the construction of BBa_K2032002. It contains the coding sequence for MDH2 which oxidizes methane to formaldehyde. It also contains the lac + pL promoter (BBa_R0011) which is repressed by LacI and induced by IPTG.</p>
 +
</div>
 +
<div class="profiles composite">
 +
<strong><a target="_blank" href="http://parts.igem.org/Part:BBa_K2032006">Fructose with RFP (BBa_K2032006)</strong></a>
 +
<img src="https://static.igem.org/mediawiki/2016/b/b3/T--UMaryland--BBa_K2032006_linear.jpg" class="linear" />
 +
<img src="https://static.igem.org/mediawiki/2016/f/f9/T--UMaryland--BBa_K2032006_plasmid.jpg" class="plasmid" />
 +
<p>Fructose construct with RFP reporter</p>
 +
</div>
 +
<div class="profiles basic">
 +
<strong><a target="_blank" href="http://parts.igem.org/Part:BBa_K2032009">MMOY/Z+TT (BBa_K2032009)</strong></a>
 +
<img src="https://static.igem.org/mediawiki/2016/4/44/T--UMaryland--BBa_K2032009_plasmid.jpg" class="plasmid" />
 +
<p>This biobrick contains the coding regions for the Y and Z subunits of the sMMO enzyme. Each coding region is preceded by a ribosome binding site (BBa_B0032) the whole biobrick is flanked by the standard iGEM double terminator (BBa_B0015
 +
)</p>
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</div>
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</div>
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<!-- <table id="table-selector"> Table of navigational links
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<thead>
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<th class="borderRight navigator borderTop" id="th-purpose" data-select="purpose"><p>Purpose</p></th>
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<th class="borderBoth navigator borderTop" id="th-background" data-select="background"><p>Background</p></th>
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<th class="borderBoth navigator borderTop" id="th-experiment" data-select="experiment"><p>Experiment</p></th>
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</thead>
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</table> -->
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<!--<div class="longText">
 +
<h4></h4>
 +
<p>Three different constructs were synthesized by UMaryland iGEM in hopes of being co-cultured. They were:</p>
 +
<ul>
 +
<li>sMMO Construct: oxidizing methane into methanol using oxygen</li>
 +
<li>Formate Construct: further oxidizing methanol into carbon dioxide using NADH</li>
 +
<li>Fructose Construct: further oxidizing methanol into cellular metabolites</li>
 +
</ul>
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</div>
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<div class="longText" id="div-protocol">
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<h4>Protocol</h4>
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</div>
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<div class="longText">
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<h4>Results</h4>
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<div id="div-results">
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</div>
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<div>
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<img class="figure" src="https://static.igem.org/mediawiki/2016/a/ae/T--UMaryland--absorbance.JPG">
 +
<small class="caption">Figure 1. Cell growth versus absorbance.</small>
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Latest revision as of 01:04, 20 October 2016

</div> </div> Modeling

Parts Collection
BioBrick Devices Submitted to the Registry
Furthering collaboration and standardization of genetic parts

UMaryland iGEM submitted various basic and composite parts to the BioBrick Registry, which aims to increase standardization in synthetic biology by allowing genes to be added together easily. We synthesized the genes, put them inside standard BioBrick plasmids, and then characterized our parts.

View:

Basic Parts Composite Parts

Fructose Pathway (BBa_K2032001)

This composite biobrick part is a combination of the coding regions for three separate enzymes involved in the metabolization of methanol. Each coding region is preempted by a ribosome binding site in order to help counteract some of the translational issues associated with polycistronic mRNA. A lacI regulated promoter that allows for induction with IPTG was included in this construct to allow for selective gene expression along with the standard iGEM double terminator.

The three enzymes encoded by this part are methanol dehydrogenase 2 (MDH), 3-hexulose-6-phosphate synthase (HPS), and 6-Phospho-3-hexuloisomerase (PHI). These enzymes serve to function as a three step pathway in which methanol is metabolized. MDH converts methanol to formaldehyde, producing a molecule of NADH in the process. Formaldehyde is then combined with a molecule of D-ribulose-5-phosphate taken from the pentose phosphate pathway to form one molecule of D-arabino-3-hexulose-6-phosphate via the usage of HPS. PHI then converts D-arabino-3-hexulose-6-phosphate to D-fructose-6-phosphate which can then undergo glycolysis.

Formate Pathway (BBa_K2032002)

The Formate Pathway is a three enzyme pathway that begins with methanol and NAD+ as substrates, and culminates in the production of NADH molecules and carbon dioxide. This pathway is used to detoxify alcohols in the cellular environment. The pathway consists of a series of oxidations: methanol oxidized to formaldehyde by Methanol Dehydrogenase 2 (MDH2); formaldehyde oxidized to formate by Formaldehyde Dehydrogenase (FALDH); and finally formate oxidized to carbon dioxide by Formate Dehydrogenase (FDH). Each of these catalyzed reactions results in lower energy products than reactants, so every reaction is coupled to the production of one NADH molecule, which contributes to energy for the cell and biomass production.

The first enzyme of the pathway, MDH2, has a low binding specificity and will oxidize many primary alcohols. The original use of this plasmid was to detoxify methanol as part of a larger pathway that consisted of eliminating methane gas from the atmosphere. The pathway has potential uses for detoxifying alcohols in the environment as well. As methanol becomes an increasingly popular liquid fuel source, one could imagine methanol spills in the future that require detoxification. This part contains one promoter that is IPTG inducible. The genes will be transcribed as a polycistronic mRNA strand. Each of the genes has a medium strength ribosome binding site. Modeling of this pathway has revealed that no toxic substrates should be produced when this pathway is expressed. The kinetics for each enzyme exist in such a way that there will be no buildup of formaldehyde of formic acid in the cell. Theoretically, this pathway should increase methanol resistance to cells that express it.

Codon optimized MDH2 with Lac/pL promoter (BBa_K2032003)

This is an intermediate used in the construction of BBa_K2032002. It contains the coding sequence for MDH2 which oxidizes methane to formaldehyde. It also contains the lac + pL promoter (BBa_R0011) which is repressed by LacI and induced by IPTG.

Fructose with RFP (BBa_K2032006)

Fructose construct with RFP reporter

MMOY/Z+TT (BBa_K2032009)

This biobrick contains the coding regions for the Y and Z subunits of the sMMO enzyme. Each coding region is preceded by a ribosome binding site (BBa_B0032) the whole biobrick is flanked by the standard iGEM double terminator (BBa_B0015 )