Team:Uppsala/signal-peptide

From 2013.igem.org

(Difference between revisions)
 
(24 intermediate revisions not shown)
Line 1: Line 1:
-
{{Team:Uppsala/standard-style}}
+
{{Team:Uppsala/signal-style}}
 +
{{Team:Uppsala/standard-style-no-frame}}
Line 12: Line 13:
</style>
</style>
 +
 +
<link href="https://2013.igem.org/Team:Uppsala/lightbox-css-code.css?action=raw&ctype=text/css" type="text/css" rel="stylesheet">
 +
 +
<script src="https://2013.igem.org/Team:Uppsala/jquery-code.js?action=raw&ctype=text/javascript" type="text/javascript"></script>
 +
 +
<script src="https://2013.igem.org/Team:Uppsala/lightbox-code.js?action=raw&ctype=text/javascript" type="text/javascript"></script>
</head>
</head>
Line 26: Line 33:
<!-- Contains sub-logo picture -->
<!-- Contains sub-logo picture -->
-
<div id="igem_logo"><a href="https://2013.igem.org/Main_Page"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/5/59/IGEMlogo.png"></a></div>
+
<div id="igem_logo"><a href="https://2013.igem.org/Team:Uppsala"><img class="igem_logo" src="https://static.igem.org/mediawiki/2013/e/e0/Uppsala2013_IGEMlogo.png"></a></div>
-
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/9/9c/Uppsala_IGEM_log_blue.png"></a></div>
+
<div id="igem"><a href="https://2013.igem.org/Main_Page"><img class="igem" src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_Uppsala_IGEM_log_blue.png"></a></div>
 +
 
 +
<img class="slogan" src="https://static.igem.org/mediawiki/2013/a/ad/Uppsala2013_Header.png">
</div>  
</div>  
Line 43: Line 52:
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a>
<li><a href="https://2013.igem.org/Team:Uppsala/project" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/a/aa/Uppsala2013_Project.png"></a>
<ul>
<ul>
-
<li><a href="https://2013.igem.org/Team:Uppsala/overview">Overview</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a>
<li><a href="https://2013.igem.org/Team:Uppsala/chassi">Chassi</a>
<ul>
<ul>
Line 49: Line 58:
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/promoters">Promoters</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/reporter-genes">Reporter genes</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/affinity-tags">Toxin-antitoxin system</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/toxin-antitoxin-system">Toxin-antitoxin system</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/vectors">Vectors</a></li>
                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li>
                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/signal-peptide">Signal peptide</a></li>
Line 56: Line 65:
                                                 <li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a>
                                                 <li><a href="https://2013.igem.org/Team:Uppsala/metabolic-engineering">Metabolic engineering</a>
                                                     <ul>
                                                     <ul>
-
                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">P-coumaric acid</a></li>
+
                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/p-coumaric-acid">p-Coumaric acid</a></li>
                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li>
                                                                 <li><a href="https://2013.igem.org/Team:Uppsala/resveratrol">Resveratrol</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/lycopene">Lycopene</a></li>
Line 62: Line 71:
                                                                 <li><a  
                                                                 <li><a  
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li>
href="https://2013.igem.org/Team:Uppsala/saffron">Saffron</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/astraxantin">Astaxanthin</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/astaxanthin">Astaxanthin</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxantin">Zeaxanthin</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/zeaxanthin">Zeaxanthin</a></li>
</ul></li>
</ul></li>
</li>  
</li>  
Line 69: Line 78:
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li>
href="https://2013.igem.org/Team:Uppsala/miraculin">Miraculin</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/chromoproteins">Chromoproteins</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/safety">Safety experiment</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/safety-experiment">Safety experiment</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/result">Result</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/results">Results</a></li>
Line 77: Line 86:
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a>
<li><a href="https://2013.igem.org/Team:Uppsala/modeling" id="list_type1"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/6/63/Uppsala2013_Modeling.png"></a>
<ul>
<ul>
-
<li><a href="https://2013.igem.org/Team:Uppsala/resveratrol-pathway">Resveratrol pathway </a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/P-Coumaric-acid-pathway">Kinetic model</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/modeling-tutorial">Modeling tutorial </a></li>
 +
 +
<li><a href="https://2013.igem.org/Team:Uppsala/toxicity-model">Toxicity model</a></li>
</ul></li>
</ul></li>
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/parts" id="list_type2"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/e/eb/Uppsala2013_parts.png"></a></li>
Line 87: Line 98:
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/carotenoid-group">Carotenoid group</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li>
<li><a href="https://2013.igem.org/Team:Uppsala/chassi-group">Chassi group</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li>
+
                                                <li><a href="https://2013.igem.org/Team:Uppsala/advisors">Advisors</a></li>
 +
</ul></li>
</ul></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/hp" id="list_type3"><img class="nav-text"      src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a>
+
<li><a href="https://2013.igem.org/Team:Uppsala/human-practice" id="list_type3"><img class="nav-text"      src="https://static.igem.org/mediawiki/2013/b/b8/Uppsala2013_HP.png"></a>
<ul>
<ul>
-
<li><a href="https://2013.igem.org/Team:Uppsala/Yoghurtproducts">Yoghurt products</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/yoghurt">Yoghurt +</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBio day</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/synbioday">SynBioDay</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/safety">Bioethics and safety</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/biosafety-and-ethics">Biosafety and ethics</a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/society">Society and outreach</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/public-opinion">Public opinion </a></li>
-
<li><a href="https://2013.igem.org/Team:Uppsala/highschool-outreach">High school outreach</a></li>
+
                                                <li><a href="https://2013.igem.org/Team:Uppsala/Outreach">High school & media </a></li>
-
                                                <li><a href="https://2013.igem.org/Team:Uppsala/outreach">Media outreach</a></li>
+
<li><a href="https://2013.igem.org/Team:Uppsala/bioart">BioArt</a></li>
 +
<li><a href="https://2013.igem.org/Team:Uppsala/LactonutritiousWorld">A LactoWorld</a></li>
 +
<li><a href="https://2013.igem.org/Team:Uppsala/killswitches">Killswitches</a></li>
 +
<li><a href="https://2013.igem.org/Team:Uppsala/realization">Patent</a></li>
</ul></li>
</ul></li>
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a></li>  
<li><a href="https://2013.igem.org/Team:Uppsala/attribution" id="list_type4"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/5/5d/Uppsala2013_Attributions.png"></a></li>  
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a>
<li><a href="https://2013.igem.org/Team:Uppsala/notebook" id="list_type3"><img class="nav-text" src="https://static.igem.org/mediawiki/2013/3/36/Uppsala2013_Notebook.png"></a>
                                     <ul>
                                     <ul>
-
                                         <li><a href="https://2013.igem.org/Team:Uppsala/protocol">Protocol</a></li>
+
                                         <li><a href="https://2013.igem.org/Team:Uppsala/safety-form">Safety form</a></li>
-
                                         <li><a href="https://2013.igem.org/Team:Uppsala/notebook">Lab journal</a></li>
+
                                         <li><a href="https://2013.igem.org/Team:Uppsala/protocols">Protocols</a></li>
                                     </ul></li>
                                     </ul></li>
</ul>
</ul>
-
</div>
+
</div>  
</div>
</div>
-
 
<div id="main_content"> <!-- Put content here -->
<div id="main_content"> <!-- Put content here -->
 +
<h1 class="main-title-left">  Signal peptides </h1>
 +
<img class="symbol-pic" src="https://static.igem.org/mediawiki/2013/4/40/Uppsala2013_Signal_peptide_symbol.png">
 +
 +
<div id="clear"></div>
 +
 +
<h1> How signal peptides work </h1>
 +
<p>A signal peptide, also called leader peptide, is the first part of extracellular proteins and can vary in length, its a short sequence fused to the N-terminus of a protein. The signal peptides we work with are 25 and 50 amino acids in length. The signal peptides enables the protein to be translocated through the bacterial plasma membrane via the SecY complex.<sup><a href="#l1">[1]</a></sup></p> 
 +
 +
<a id="b1"></a>
 +
<h1> Lactobacillus - A champion of protein secretion </h1>
 +
<p>One of the specific advantageous traits of the lactobacillus genus is that they are great at secreting proteins. Therefore we decided to synthesize the signal peptide usp45 from lactococcus lactis according to the Freiburg fusion standard to enable fusion with any protein. This peptide has been shown to work in lactobacillus reuteri together with GFP and will probably work in other lactobacillus species.<sup><a href="#l2">[2]</a></sup></p>
 +
 +
<a href="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_signal-peptides.jpg" data-lightbox="roadtrip"><img class="method-plasmid" src="https://static.igem.org/mediawiki/2013/9/9d/Uppsala2013_signal-peptides.jpg"></a>
 +
 +
<p>One of the goals in our project was to make a probiotic bacteria express the protein miraculin (Mir in picture) which would work as a sweetener. However because it works by binding to receptors on the tongue and the protein is too big to pass through the cell membrane of the bacteria it needs to be actively secreted to work optimally. Fusing a signal peptide to the N-terminus of miraculin can solve this. We have synthesised miraculin fused together with usp45. Because we received the synthesised miraculin gene late, we did not have time to put it in pSB1C3 and send it to the registry. We have also not yet had the chance to transform and characterise it in Lactobacillus.</p>
 +
 +
<h1> References: </h1>
 +
<a name="l1">[1]</a> Rapoport T. (Nov. 2007). "Protein translocation across the eukaryotic endoplasmic reticulum and bacterial plasma membranes.". Nature 450 (7170): 663–9.
 +
<br><br>
 +
<a name="l2">[2]</a> Chi-Ming Wu. (April 2006). “Green fluorescent protein is a reliable reporter for screening signal peptides functional in Lactobacillus reuteri” Journal of Microbiological Methods (2006) 181-186.
Line 118: Line 152:
<div id="bottom-pic">
<div id="bottom-pic">
-
<img class="bottom-pic" src="https://static.igem.org/mediawiki/2013/a/aa/Bottom_picture.png">
+
</div>
</div>

Latest revision as of 21:30, 28 October 2013

Signal peptides

How signal peptides work

A signal peptide, also called leader peptide, is the first part of extracellular proteins and can vary in length, its a short sequence fused to the N-terminus of a protein. The signal peptides we work with are 25 and 50 amino acids in length. The signal peptides enables the protein to be translocated through the bacterial plasma membrane via the SecY complex.[1]

Lactobacillus - A champion of protein secretion

One of the specific advantageous traits of the lactobacillus genus is that they are great at secreting proteins. Therefore we decided to synthesize the signal peptide usp45 from lactococcus lactis according to the Freiburg fusion standard to enable fusion with any protein. This peptide has been shown to work in lactobacillus reuteri together with GFP and will probably work in other lactobacillus species.[2]

One of the goals in our project was to make a probiotic bacteria express the protein miraculin (Mir in picture) which would work as a sweetener. However because it works by binding to receptors on the tongue and the protein is too big to pass through the cell membrane of the bacteria it needs to be actively secreted to work optimally. Fusing a signal peptide to the N-terminus of miraculin can solve this. We have synthesised miraculin fused together with usp45. Because we received the synthesised miraculin gene late, we did not have time to put it in pSB1C3 and send it to the registry. We have also not yet had the chance to transform and characterise it in Lactobacillus.

References:

[1] Rapoport T. (Nov. 2007). "Protein translocation across the eukaryotic endoplasmic reticulum and bacterial plasma membranes.". Nature 450 (7170): 663–9.

[2] Chi-Ming Wu. (April 2006). “Green fluorescent protein is a reliable reporter for screening signal peptides functional in Lactobacillus reuteri” Journal of Microbiological Methods (2006) 181-186.