Team:UniSalento Lecce

From 2013.igem.org

(Difference between revisions)
 
(47 intermediate revisions not shown)
Line 1: Line 1:
<html>
<html>
<head>
<head>
-
 
+
<meta http-equiv="Content-Type" content="text/html; charset=UTF-8" />
-
<meta http-equiv="Content-Type" content="text/html; charset=UTF-8" />
+
<link rel="stylesheet" href="https://2013.igem.org/Team:UniSalento_Lecce/css/common.css?action=raw&ctype=text/css" type="text/css" />
-
        <link rel="stylesheet" href="https://2013.igem.org/Team:UniSalento_Lecce/css/common.css?action=raw&ctype=text/css" type="text/css" />
+
<link rel="stylesheet" href="https://2013.igem.org/Team:UniSalento_Lecce/css/slider.css?action=raw&ctype=text/css" type="text/css" />
-
        <link rel="stylesheet" href="https://2013.igem.org/Team:UniSalento_Lecce/css/slider.css?action=raw&ctype=text/css" type="text/css" />
+
<script src="https://2013.igem.org/Team:UniSalento_Lecce/js/go-to-top.js?action=raw&ctype=text/javascript"></script>
-
<script src="https://2013.igem.org/Team:UniSalento_Lecce/js/go-to-top.js?action=raw&ctype=text/javascript"></script>
+
<script src="https://2013.igem.org/Team:UniSalento_Lecce/js/fixed-navbar.js?action=raw&ctype=text/javascript"></script>
-
<script src="https://2013.igem.org/Team:UniSalento_Lecce/js/fixed-navbar.js?action=raw&ctype=text/javascript"></script>
+
<script src="https://2013.igem.org/Team:UniSalento_Lecce/js/jquery.carouFredSel-5.6.1.js?action=raw&ctype=text/javascript"></script>
-
        <script src="https://2013.igem.org/Team:UniSalento_Lecce/js/jquery.carouFredSel-5.6.1.js?action=raw&ctype=text/javascript"></script>
+
<script src="https://2013.igem.org/Team:UniSalento_Lecce/js/slider.js?action=raw&ctype=text/javascript"></script>
-
        <script src="https://2013.igem.org/Team:UniSalento_Lecce/js/slider.js?action=raw&ctype=text/javascript"></script>
+
<script type="text/javascript">
<script type="text/javascript">
-
    $(document).ready(function() {
+
$(document).ready(function() {
-
        $("p").filter( function() {
+
$("p").filter( function() {
-
            return $.trim($(this).html()) == '';
+
return $.trim($(this).html()) == '';
-
        }).remove()
+
}).remove()
-
    });
+
});
</script>
</script>
-
 
</head>
</head>
<body>
<body>
<!-- ****************** Start Header *********************** -->
<!-- ****************** Start Header *********************** -->
-
<header></header>
+
<header>
 +
<img src="https://static.igem.org/mediawiki/2013/0/0b/Unisalento_logo_team.png" height="110px" style="position: absolute; left:37px; top:105px">
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce" style="background: none">
 +
<img src="https://static.igem.org/mediawiki/2013/a/a5/Unisalento_logo_project.png" width="450px" style="margin-top:0px;margin-left:0px">
 +
</a>
 +
<a href="http://www.igem.org" style="background: none">
 +
<img src="https://static.igem.org/mediawiki/2013/b/b0/Unisalento_Igem_official.png" height="75px" style="position: absolute; right:40px; top:15px">
 +
</a>
 +
<a href="https://www.unisalento.it" style="background: none">
 +
<img src="https://static.igem.org/mediawiki/2013/c/c7/Unisalento_logo_university.png" height="85px" style="position: absolute; right:40px; top:125px">
 +
</a>
 +
</header>
<!-- ****************** End Header ************************* -->
<!-- ****************** End Header ************************* -->
<!-- ****************** Start Navigation Bar ************************* -->
<!-- ****************** Start Navigation Bar ************************* -->
<nav>
<nav>
-
<ul id="css3menu1" class="topmenu">
+
<ul id="css3menu1" class="topmenu">
-
<li class="topmenu"><a href="#" title="Home" style="height:24px;line-height:24px;"><img src="https://static.igem.org/mediawiki/2013/b/bc/Unisalento_home.png" alt="">Home</a></li>
+
<li class="topmenu">
-
<li class="toproot"><a href="team.html" title="Team" style="height:24px;line-height:24px;"><img src="https://static.igem.org/mediawiki/2013/a/ab/Users.png" alt="">Team</a>
+
<a href="#" title="Home" style="height:24px;line-height:24px;">
-
<div class="submenu" style="width:230px">
+
<img src="https://static.igem.org/mediawiki/2013/b/bc/Unisalento_home.png" alt="">
-
<div class="column" style="width:100%">
+
Home
-
<ul>
+
</a>
-
<li><a href="team.html" title="Team"><img src="https://static.igem.org/mediawiki/2013/1/12/Unisalento_user1.png" alt="">Team</a></li>
+
</li>
-
<li><a href="university.html" title="University/Department"><img src="https://static.igem.org/mediawiki/2013/c/c3/Unisalento_university.png" alt="">University/Department</a></li>
+
<li class="toproot">
-
</ul>
+
<a href="#" title="Team" style="height:24px;line-height:24px;">
-
</div>
+
<img src="https://static.igem.org/mediawiki/2013/5/50/Unisalento_users.png" alt="">
-
</div>
+
Team
-
</li>
+
</a>
-
<li class="toproot">
+
<div class="submenu" style="width:230px">
-
<a href="#" title="Project" style="height:24px;line-height:24px;"><span><img src="https://static.igem.org/mediawiki/2013/d/d4/Bag.png" alt="">Project</span></a>
+
<div class="column" style="width:100%">
-
<div class="submenu" style="width:250px">
+
<ul>
-
<div class="column" style="width:100%">
+
<li>
-
<ul>
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Team" title="Team">
-
<li><a href="#" title="Why? ... An overview"><img src="https://static.igem.org/mediawiki/2013/3/30/Question.png" alt="">Why? ... An overview</a></li>
+
<img src="https://static.igem.org/mediawiki/2013/1/12/Unisalento_user1.png" alt="">
-
<li><a href="#" title="Experimental data and results"><img src="https://static.igem.org/mediawiki/2013/7/75/Unisalento_graph.png" alt="">Experimental data and results</a></li>
+
Team
-
<li>
+
</a>
-
<a href="#" title="Biobricks"><span><img src="https://static.igem.org/mediawiki/2013/c/cc/Unisalento_bricks.png" alt="">Biobricks</span></a>
+
</li>
-
<div class="submenu" style="width:120px;">
+
<li>
-
<div class="column" style="width:100%">
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/University" title="University/Department">
-
<ul>
+
<img src="https://static.igem.org/mediawiki/2013/c/c3/Unisalento_university.png" alt="">
-
<li><a href="#" title="data 1"><img src="https://static.igem.org/mediawiki/2013/7/7d/Data.png" alt="">data 1</a></li>
+
University/Department
-
<li><a href="#" title="data 2"><img src="https://static.igem.org/mediawiki/2013/7/7d/Data.png" alt="">data 2</a></li>
+
</a>
-
<li><a href="#" title="data 3"><img src="https://static.igem.org/mediawiki/2013/7/7d/Data.png" alt="">data 3</a></li>
+
</li>
-
<li><a href="#" title="data 4"><img src="https://static.igem.org/mediawiki/2013/7/7d/Data.png" alt="">data 4</a></li>
+
</ul>
-
<li><a href="#" title="data 5"><img src="https://static.igem.org/mediawiki/2013/7/7d/Data.png" alt="">data 5</a></li>
+
</div>
-
</ul>
+
</div>
-
</div>
+
</li>
-
</div>
+
<li class="toproot">
-
</li>
+
<a href="#" title="Project" style="height:24px;line-height:24px;">
-
</ul>
+
<span>
-
</div>
+
<img src="https://static.igem.org/mediawiki/2013/f/f2/Unisalento_project.png" alt="">
-
</div>
+
Project
-
</li>
+
</span>
-
<li class="topmenu">
+
</a>
-
<a href="#" title="Model" style="height:24px;line-height:24px;"><span><img src="https://static.igem.org/mediawiki/2013/a/a4/Idea.png" alt="">Model</span></a>
+
<div class="submenu" style="width:250px">
-
</li>
+
<div class="column" style="width:100%">
-
<li class="toproot">
+
<ul>
-
<a href="#" title="Nootebook" style="height:24px;line-height:24px;"><span><img src="https://static.igem.org/mediawiki/2013/5/51/Notes.png" alt="">Notebook</span></a>
+
<li>
-
<div class="submenu" style="width:200px;">
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Overview" title="Why NickBusters?">
-
<div class="column" style="width:100%">
+
<img src="https://static.igem.org/mediawiki/2013/7/79/Unisalento_question.png" alt="">
-
<ul>
+
Why NickBusters?
-
<li><a href="notebook.html" title="Weekly Journal"><img src="https://static.igem.org/mediawiki/2013/e/e0/Date.png" alt="">Weekly Journal</a></li>
+
</a>
-
<li><a href="#" title="Protocols"><img src="https://static.igem.org/mediawiki/2013/3/3b/Db.png" alt="">Protocols</a></li>
+
</li>
-
</ul>
+
<li>
-
</div>
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Data" title="Experimental data and results">
-
</div>
+
<img src="https://static.igem.org/mediawiki/2013/c/c9/Unisalento_chart.png" alt="">
-
</li>
+
Experimental data and results
-
<li class="topmenu">
+
</a>
-
<a href="#" title="Safety" style="height:24px;line-height:24px;"><span><img src="https://static.igem.org/mediawiki/2013/5/5e/Unisalento_safety.png" alt="">Safety</span></a>
+
</li>
-
</li>
+
<li>
-
<li class="toproot"><a href="#" title="Extra" style="height:24px;line-height:24px;"><img src="https://static.igem.org/mediawiki/2013/2/2e/Star.png" alt="">Extra</a>
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Parts" title="Biobricks and parts">
-
<div class="submenu" style="width:220px;">
+
<img src="https://static.igem.org/mediawiki/2013/3/36/Unisalento_lego.png" alt="">
-
<div class="column" style="width:100%">
+
Biobricks and parts
-
<ul>
+
</a>
-
<li><a href="#" title="Human_practise"><img src="https://static.igem.org/mediawiki/2013/3/39/World.png" alt="">Human practicies</a></li>
+
</li>
-
<li><a href="#" title="Sposor"><img src="https://static.igem.org/mediawiki/2013/7/76/Flag.png" alt="">Sponsor</a></li>
+
<li>
-
<li><a href="#" title="Acknowledgment"><img src="https://static.igem.org/mediawiki/2013/9/95/Search.png" alt="">Acknowledgment</a></li>
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Applications" title="Applications">
-
</ul>
+
<img src="https://static.igem.org/mediawiki/2013/6/6e/Unisalento_applications.png" alt="">
-
</div>
+
Applications
-
</div>
+
</a>
-
</li>
+
</li>
-
</ul>
+
</ul>
-
<!-- ****************** Start Social Links ************************ -->
+
</div>
-
<div id="social_facebook"><a href="https://www.facebook.com/IgemUnisalento2013" style="background: none"><img src="https://static.igem.org/mediawiki/2013/f/f3/Unisalento_facebook.png" alt="" height="50px"></a></div>
+
</div>
-
<div id="social_twitter"><a href="https://www.twitter.com/iGEM_UniSalento" style="background: none"><img src="https://static.igem.org/mediawiki/2013/7/7d/Unisalento_twitter.png" alt="" height="50px"></a></div>
+
</li>
-
<div id="social_gmail"><a href="http://www.google.com"><img src="https://static.igem.org/mediawiki/2013/d/d3/Unisalento_gmail.png" alt="" height="50px"></a></div>
+
<li class="topmenu">
-
<div class="clear"></div>
+
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Human_Practices" title="Human practices" style="height:24px;line-height:24px;">
-
<!-- ******************* End Social Links ************************* -->
+
<span>
 +
<img src="https://static.igem.org/mediawiki/2013/6/6b/Unisalento_world.png" alt="">
 +
Human practices
 +
</span>
 +
</a>
 +
</li>
 +
<li class="toproot">
 +
<a href="#" title="Nootebook" style="height:24px;line-height:24px;">
 +
<span>
 +
<img src="https://static.igem.org/mediawiki/2013/2/2d/Unisalento_notes.png" alt="">
 +
Notebook
 +
</span>
 +
</a>
 +
<div class="submenu" style="width:200px;">
 +
<div class="column" style="width:100%">
 +
<ul>
 +
<li>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Notebook" title="Weekly Journal">
 +
<img src="https://static.igem.org/mediawiki/2013/3/34/Unisalento_date.png" alt="">
 +
Weekly Journal
 +
</a>
 +
</li>
 +
<li>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Protocols" title="Protocols">
 +
<img src="https://static.igem.org/mediawiki/2013/c/c5/Unisalento_db.png" alt="">
 +
Protocols
 +
</a>
 +
</li>
 +
</ul>
 +
</div>
 +
</div>
 +
</li>
 +
<li class="topmenu">
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Safety" title="Safety" style="height:24px;line-height:24px;">
 +
<span>
 +
<img src="https://static.igem.org/mediawiki/2013/5/5e/Unisalento_safety.png" alt="">
 +
Safety
 +
</span>
 +
</a>
 +
</li>
 +
<li class="toproot">
 +
<a href="#" title="Extra" style="height:24px;line-height:24px;">
 +
<img src="https://static.igem.org/mediawiki/2013/3/32/Unisalento_star.png" alt="">
 +
Extra
 +
</a>
 +
<div class="submenu" style="width:220px;">
 +
<div class="column" style="width:100%">
 +
<ul>
 +
<li>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Modeling" title="Model">
 +
<img src="https://static.igem.org/mediawiki/2013/9/9d/Unisalento_model.png" alt="">
 +
Model
 +
</a>
 +
</li>
 +
<li>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Sponsors" title="Sposors">
 +
<img src="https://static.igem.org/mediawiki/2013/f/f7/Unisalento_flag.png" alt="">
 +
Sponsors
 +
</a>
 +
</li>
 +
<li>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Attributions" title="Acknowledgments">
 +
<img src="https://static.igem.org/mediawiki/2013/e/e9/Unisalento_search.png" alt="">
 +
Acknowledgments
 +
</a>
 +
</li>
 +
</ul>
 +
</div>
 +
</div>
 +
</li>
 +
</ul>
 +
<!-- ****************** Start Social Links ************************ -->
 +
<div id="social_facebook"><a href="https://www.facebook.com/IgemUnisalento2013" style="background: none"><img src="https://static.igem.org/mediawiki/2013/f/f3/Unisalento_facebook.png" alt="" height="50px"></a></div>
 +
<div id="social_twitter"><a href="https://www.twitter.com/iGEM_UniSalento" style="background: none"><img src="https://static.igem.org/mediawiki/2013/7/7d/Unisalento_twitter.png" alt="" height="50px"></a></div>
 +
<div id="social_gmail"><a href="https://plus.google.com/u/0/108724165022727316894" style="background: none"><img src="https://static.igem.org/mediawiki/2013/d/d3/Unisalento_gmail.png" alt="" height="50px"></a></div>
 +
<div class="clear"></div>
 +
<!-- ******************* End Social Links ************************* -->
</nav>
</nav>
<!-- ****************** End Navigation Bar ************************* -->
<!-- ****************** End Navigation Bar ************************* -->
<!-- ********************************************** Start Main **************************************************** -->
<!-- ********************************************** Start Main **************************************************** -->
<div id="main">
<div id="main">
 +
<div id="main_wrap">
 +
<!-- ************************************** Start Slider ***************************************** -->
 +
<div id="slider">
 +
<div id="bg"></div>
 +
<div id="carousel">
 +
<div>
 +
<img class="img-front" src="https://static.igem.org/mediawiki/2013/8/88/Unisalento_slider1.jpg" alt="" />
 +
<img class="img-back" src="https://static.igem.org/mediawiki/2013/a/af/Unisalento_slider2.jpg" alt="" />
 +
<h3>Sense it!</h3>
 +
<p style="font-size: 14px">Population #1 feels the danger and calls up for help! When sensing nickel in the environment, this population turns red, then releases a QS molecule, recruiting population 2! </p>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Overview#o2">Read more &raquo;</a>
 +
</div>
 +
<div>
 +
<img class="img-front" src="https://static.igem.org/mediawiki/2013/5/52/Unisalento_slider3.jpg" alt="" />
 +
<img class="img-back" src="https://static.igem.org/mediawiki/2013/2/24/Unisalento_slider4.jpg" alt=""  />
 +
<h3>Store it!</h3>
 +
<p style="font-size: 14px">Population #2 receives the QS signal, which activates the adsorption of nickel; then amplifies the QS signal, inducing further bacteria to accumulate the heavy metal. </p>
 +
<a href="https://2013.igem.org/Team:UniSalento_Lecce/Overview#o3">Read more &raquo;</a>
 +
</div>
 +
</div>
 +
<a id="prev" href="#"><span></span></a>
 +
<a id="next" href="#"><span></span></a>
 +
</div>
 +
<!-- ************************************** End Slider ******************************************* -->
 +
<div id="container">
 +
<img src="https://static.igem.org/mediawiki/2013/6/68/Sticker02.png" alt="" height="60px" style="margin-left: -111px; margin-top: -30px; -moz-margin-start: -658px; position: absolute">
 +
<!-- *************************************************************************************************** -->
 +
<div class="split2_3l">
 +
<br/>
 +
<br/>
 +
<img src="https://static.igem.org/mediawiki/2013/1/1a/Unisalento.becker.png" width="547px">
 +
<h2>Introduction to NickBusters Project</h2>
 +
<p>
 +
Nickel is one of the most widespread heavy metals in the ecosystem, and it is essential for many organisms,
 +
from bacteria to higher forms of life. However, as all the heavy metals, even a small amount of Nickel higher
 +
than the essential dose could be toxic, leading to various noxious effects <a href="#biblio" style="color: #8ac101">[1-2]</a>: then the need to remove its
 +
excess from many substrates. Nowadays <b>bacteria-mediated bioremediation</b> from inorganic substances seems to be a considerably relevant frontier in microbial biotechnologies <a href="#biblio" style="color: #8ac101">[3]</a>.
 +
Our project aims to develop a living system, in an easily monitorable bacterial platform, who would work as a
 +
<b>Nickel detector</b> and a <b>Nickel remediation</b> system. The whole system is based on genetic parts from the
 +
pathogen <i>Helicobacter pylori</i>, whose evolution led to a stronger reliance on Nickel ions for host interaction
 +
and pathogenesis.
 +
<b>Sensing</b> relies on nickel regulator <i>NikR</i> from <i>H. pylori</i> (<i>HpNikR</i>): though homolog of <i>E.coli NikR</i>, it has a
 +
stronger dependence on Nickel ions and a wider range of regulated elements, being a rare case of pleiotropic
 +
regulator in bacteria.
 +
As <i>HpNikR</i> could contemporaneously activate and repress gene expression <a href="#biblio" style="color: #8ac101">[4-5-6]</a>, we thought to develop
 +
a synthetic gene cluster under <i>HpNikR</i> - regulated promoters. With a <b>double regulation</b>, one of the promoters
 +
being activated while the other gets repressed, the system allows to follow up Nickel sensing as a
 +
fluorescence signal from fluorescent proteins. Thus, adding bacteria to a nickel containing medium results in
 +
<b>switching off a basal occurring green fluorescence</b> and in <b>switching on a red fluorescence signal</b>.
 +
</p>
 +
</div>
 +
<div class="split1_3r">
 +
<h2>Twitter</h2>
 +
<a class="twitter-timeline" href="https://twitter.com/iGEM_UniSalento" data-widget-id="356106138686722049">Tweets di @iGEM_UniSalento</a>
 +
<script>!function(d,s,id){var js,fjs=d.getElementsByTagName(s)[0],p=/^http:/.test(d.location)?'http':'https';if(!d.getElementById(id)){js=d.createElement(s);js.id=id;js.src=p+"://platform.twitter.com/widgets.js";fjs.parentNode.insertBefore(js,fjs);}}(document,"script","twitter-wjs");</script>
 +
</div>
 +
<div class="clear"></div>
 +
<!-- *************************************************************************************************** -->
 +
<div class="nosplit">
 +
<img src="https://static.igem.org/mediawiki/2013/1/1d/Unisalento_welcome1.png" alt="" width="100%">
 +
<br/>
 +
<p>
 +
The Nickel <b>storage</b> system is even taken from our pathogenic friend: <i>Hpn</i> protein is a Nickel-binding protein,
 +
whose purpose is to store the Nickel ions inside the cell and to supply them with other protein interactors, from
 +
Nickel dependent heat shock proteins to pathogenesis factors as urease <a href="#biblio" style="color: #8ac101">[7]</a>. <i>Hpn</i> can be expressed in a
 +
heterologous model system (<i>E.coli</i>), increasing bacterial resistance up to a toxic dose of Nickel ions and allowing
 +
Ni++ intracellular accumulation <a href="#biblio" style="color: #8ac101">[8]</a>.
 +
We thought to split the <b>sensing and storing machineries into two different bacterial populations</b>, making them
 +
communicate with each other through Quorum Sensing <a href="#biblio" style="color: #8ac101">[9]</a>: we used the <i>LuxI/LuxR</i> system from <i>Alivibrio fischerii</i>. The first
 +
population senses nickel concentration in medium ( and translates it in a fluorescence signal), then activating
 +
<i>Acyl-Homoserine-Lactones</i> (<i>AHLs</i>) production. The second population receives the <i>QS</i> signal and sums it up to the nickel detection signal (as in an AND gate), activating <i>Hpn</i> (the storage protein) expression and <i>AHLs</i> signal amplification.
 +
</p>
 +
<br/>
 +
<img src="https://static.igem.org/mediawiki/2013/6/68/Unisalento_welcome2.png" alt="" width="100%">
 +
<br/>
 +
<br/>
 +
</div>
 +
<div class="clear"></div>
 +
<!-- *************************************************************************************************** -->
 +
<img src="https://static.igem.org/mediawiki/2013/2/2f/Unisalento_dna.png" alt="" style="margin-bottom: -100px; margin-left: 360px; position: absolute">
 +
<!-- *************************************************************************************************** -->
 +
</div>
 +
<div class="clear"></div>
 +
<!-- *************************************************************************************************** -->
 +
<div id="container">
 +
<!-- *************************************************************************************************** -->
 +
<div class="nosplit">
 +
<a name="biblio"><h2>Bibliography</h2></a>
 +
<ol>
 +
<li>Denkhaus and Salnikow, Nickel essentiality, toxicity and carcinogenity; Critical Reviews in Oncology/Hematology, 42 (2002) 35–56</li>
 +
<li>“Nickel, Elemental” on TOXNET- Databases on toxicology, hazardous chemicals, environmental health, and toxic releases.</li>
 +
<li>Gadd, Metals, minerals and microbes: geomicrobiology and bioremediation; Microbiology, 156 (2010), 609–643</li>
 +
<li>Dosanjh and Michel, Microbial nickel metalloregulation: NikRs for nickel ions; Current Opinion in Chemical Biology, 10 (2006), 123–130</li>
 +
<li>Contreras et al., Characterization of the roles of NikR, a nickel-responsive pleiotropic autoregulator of Helicobacter pylori; Molecular Microbiology, 49 (4) (2003), 947–963</li>
 +
<li>Muller et al., Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori; Nucleic Acids Research, 39 (17) (2011), 7564–7575</li>
 +
<li>Seshadri et al., Roles of His-Rich Hpn and Hpn-Like Proteins in Helicobacter pylori Nickel Physiology; Journal of Bacteriology, 189 (11) (2007), 4120–4126</li>
 +
<li>Ge et al., Expression and characterization of a histidine-rich protein, Hpn: potential for Ni2+ storage in Helicobacter pylori; Biochemistry Journal 393 (2006), 285–293</li>
 +
<li>Miller, Bassler, Quorum Sensing in bacteria; Annual Review of Microbiology 55 (2001), 165-99.</li>
 +
</ol>
 +
</div>
 +
<!-- *************************************************************************************************** -->
 +
</div>
 +
<!-- *************************************************************************************************** -->
 +
</div>
 +
</div>
 +
<div class="clear"></div>
 +
<!-- ********************* Start Footer *********************************** -->
 +
<footer>
<div id="main_wrap">
<div id="main_wrap">
-
<!-- ************************************** Start Slider ***************************************** -->
+
<div class="split2_3r" style="width:50%;text-align:right;padding-right:50px">
-
<div id="slider">
+
<img src="https://static.igem.org/mediawiki/2013/f/f9/Unisalento_igem_official.png" width="200px" align="left" style="margin-top:20px;margin-left:-45px">
-
<div id="bg"></div>
+
<h2 style="margin-right:70px;color:#cccccc">Sponsors</h2>
-
<div id="carousel">
+
<table style="float:right;background-color:transparent">
-
<div>
+
<tr>
-
<img class="img-front" src="images/slider/3.jpg" alt="" />
+
<td><a href="http://www.mps.it/" style="background: none"><img src="https://static.igem.org/mediawiki/2013/5/56/Unisalento_sponsor_mps_small.png" width="45px" align="center"/></a></td>
-
<img class="img-back" src="images/slider/4.jpg" alt="" />
+
<td><a href="http://www.mathworks.it/" style="background: none"><img src="https://static.igem.org/mediawiki/2013/1/1f/Unisalento_sponsor_mathworks_small.png" width="45px" align="center"/></a></td>
-
<h3>NICKBUSTERS</h3>
+
<td><a href="http://www.unisalentostore.it/" style="background: none"><img src="https://static.igem.org/mediawiki/2013/4/41/Unisalento_sponsor_unisalentostore_small.png" height="45px" align="right"/></a></td>
-
<p>Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. </p>
+
</tr>
-
<a href="#">Read more &raquo;</a>
+
<tr>
-
</div>
+
<td><a href="http://www.idt.com/" style="background: none"><img src="https://static.igem.org/mediawiki/2013/f/f4/Unisalento_sponsor_idt_small.png" width="45px" align="center"/></a></td>
-
<div>
+
<td><a href="https://www.neb.com/" style="background: none"><img src="https://static.igem.org/mediawiki/2013/2/25/Unisalento_sponsor_neb_small.png" height="45px" align="center"/></a></td>
-
<img class="img-front" src="images/slider/1.jpg" alt="" />
+
<td><a href="http://www.dasitsciences.it/" style="background: none"><img src="https://static.igem.org/mediawiki/2013/4/44/Unisalento_sponsor_dasit_small.png" width="45px" align="right"/></a></td>
-
<img class="img-back" src="images/slider/2.jpg" alt="" />
+
</tr>
-
<h3>Welcome...</h3>
+
</table>
-
<p>Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. </p>
+
-
<a href="#">Read more &raquo;</a>
+
-
</div>
+
-
<div>
+
-
<img class="img-front" src="images/slider/5.jpg" alt="" />
+
-
<img class="img-back" src="images/slider/6.jpg" alt="" />
+
-
<h3>More info</h3>
+
-
<p>Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. </p>
+
-
<a href="#">Read more &raquo;</a>
+
-
</div>
+
-
</div>
+
-
<a id="prev" href="#"><span></span></a>
+
-
<a id="next" href="#"><span></span></a>
+
</div>
</div>
-
<!-- ************************************** End Slider ******************************************* -->
+
<div class="split2l" style="width:30%;text-align:left;margin-left:50px">
-
<div id="container">
+
<h2 style="margin-left:40px;color:#cccccc">Contacts</h2>
-
<img src="https://static.igem.org/mediawiki/2013/6/68/Sticker02.png" alt="" height="60px" style="margin-left: -111px; margin-top: -30px; -moz-margin-start: -658px; position: absolute">
+
<table style="float:left;background-color:transparent">
-
<!-- *************************************************************************************************** -->
+
<tr>
-
<div class="split2_3l">
+
<td><a href="https://www.facebook.com/IgemUnisalento2013" style="background: none"><img src="https://static.igem.org/mediawiki/2013/a/ac/Unisalento_facebook_small.png" width="35px" align="left"/></a></td>
-
<br/>
+
<td><font style="font-size:15px;color:#cccccc">Facebook</font></td>
-
<br/>
+
</tr>
-
<p>
+
<tr>
-
Lorem ipsum dolor sit amet, consectetuer adipiscing elit, sed diam nonummy nibh euismod tincidunt ut laoreet dolore magna aliquam erat volutpat. Ut wisi enim ad minim veniam, quis nostrud exerci tation ullamcorper suscipit lobortis nisl ut aliquip ex ea commodo consequat. Duis autem vel eum iriure dolor in hendrerit in vulputate velit esse molestie consequat, vel illum dolore eu feugiat nulla facilisis at vero eros et accumsan et iusto odio dignissim qui blandit praesent luptatum zzril delenit augue duis dolore te feugait nulla facilisi.
+
<td><a href="https://www.twitter.com/iGEM_UniSalento" style="background: none"><img src="https://static.igem.org/mediawiki/2013/b/b7/Unisalento_twitter_small.png" width="35px" align="left"/></a></td>
-
</p>
+
<td><font style="font-size:15px;color:#cccccc">Twitter</font></td>
-
<img src="images/img/slide02.jpg" width="100%">
+
</tr>
-
<p>
+
<tr>
-
Investigationes demonstraverunt lectores legere me lius quod ii legunt saepius. Claritas est etiam processus dynamicus, qui sequitur mutationem consuetudium lectorum. Lorem ipsum dolor sit amet, consectetuer adipiscing elit, sed diam nonummy nibh euismod tincidunt ut laoreet dolore magna aliquam erat volutpat. Ut wisi enim ad minim veniam, quis nostrud exerci tation ullamcorper suscipit lobortis nisl ut aliquip ex ea commodo consequat. Duis autem vel eum iriure dolor in hendrerit in vulputate velit esse molestie consequat, vel illum dolore eu feugiat nulla facilisis at vero eros et accumsan et iusto odio dignissim qui blandit praesent luptatum zzril delenit augue duis dolore te feugait nulla facilisi. Nam liber tempor cum soluta nobis eleifend option congue nihil imperdiet doming id quod mazim placerat facer possim assum.
+
<td><a href="https://plus.google.com/u/0/108724165022727316894" style="background: none"><img src="https://static.igem.org/mediawiki/2013/d/d7/Unisalento_google_small.png" width="35px" align="left"/></a></td>
-
</p>
+
<td><font style="font-size:15px;color:#cccccc">Google+</font></td>
-
</div>
+
</tr>
-
<div class="split1_3r">
+
</table>
-
<h2>Twitter</h2>
+
<br><br><br><br>
-
<a class="twitter-timeline" href="https://twitter.com/iGEM_UniSalento" data-widget-id="356106138686722049">Tweets di @iGEM_UniSalento</a>
+
<p style="margin-top:20px">
-
<script>!function(d,s,id){var js,fjs=d.getElementsByTagName(s)[0],p=/^http:/.test(d.location)?'http':'https';if(!d.getElementById(id)){js=d.createElement(s);js.id=id;js.src=p+"://platform.twitter.com/widgets.js";fjs.parentNode.insertBefore(js,fjs);}}(document,"script","twitter-wjs");</script>
+
<font style="font-size:15px">
-
</div>
+
E-mail:
-
<div class="clear"></div>
+
</font>
-
<!-- *************************************************************************************************** -->
+
<a href="mailto:igem.unisalento@gmail.com" style="color:#cccccc;background:none;font-size: 15px">
-
<div class="nosplit">
+
igem.unisalento@gmail.com
-
<h2>Introduction to NickBusters Project</h2>
+
</a>
-
<p>
+
<br>
-
Nickel is one of the most widespread heavy metals in the ecosystem, and it is essential for many organisms,
+
</p>
-
from bacteria to higher forms of life. However, as all the heavy metals, even a small amount of Nickel higher
+
-
than the essential dose could be toxic, leading to various noxious effects <a href="#biblio" style="color: #8ac101">[1-2]</a>: then the need to remove its
+
-
excess from many substrates. <b>Nowadays bacteria-mediated</b> bioremediation from inorganic substances seems to be a considerably relevant frontier in microbic biotechnologies <a href="#biblio" style="color: #8ac101">[3]</a>.
+
-
Our project aims to develop a living system, in an easy monitorable bacterial platform, who would work as a
+
-
<b>Nickel detector</b> and a <b>Nickel remediation</b> system. The whole system is based on genetic parts from the
+
-
pathogen <i>Helicobacter pylori</i>, whose evolution led to a stronger reliance on Nickel ions for host interaction
+
-
and pathogenesis.
+
-
<b>Sensing</b> relies on nickel regulator <i>NikR</i> from <i>H. pylori</i> (<i>HpNikR</i>): though homolog of <i>E.coli NikR</i>, it has a
+
-
stronger dependance on Nickel ions and a wider range of regulated elements, being a rare case of pleiotropic
+
-
regulator in bacteria.
+
-
As <i>HpNikR</i> could contemporaneously activate and repress gene expression <a href="#biblio" style="color: #8ac101">[4-5-6]</a>, we thought of developing
+
-
a synthetic gene cluster under <i>HpNikR</i> - regulated promoters. With a <b>double regulation</b>, one of the promoters
+
-
being activated while the other being repressed, the system allows to follow up Nickel sensing as a
+
-
fluorescence signal from fluorescent proteins. Thus, adding bacteria to a nickel containing medium results in
+
-
<b>switching off a basal occuring green fluorescence</b> and in <b>switching on a red fluorescence signal</b>.
+
-
</p>
+
-
<br/>
+
-
<img src="https://static.igem.org/mediawiki/2013/1/1d/Unisalento_welcome1.png" alt="" width="100%">
+
-
<br/>
+
-
<p>
+
-
The Nickel <b>storage</b> system is even taken from our pathogenic friend: <i>Hpn</i> protein is a Nickel binding protein,
+
-
whose role is to store the Nickel ions inside the cell and to supply them to other proteic interactors, from
+
-
Nickel dependent heat shock proteins to pathogenesis factors as urease <a href="#biblio" style="color: #8ac101">[7]</a>. <i>Hpn</i> can be expressed in a
+
-
heterologous model system (<i>E.coli</i>), increasing bacterial resistance to toxic dose of Nickel ions and allowing
+
-
Ni++ intracellular accumulation <a href="#biblio" style="color: #8ac101">[8]</a>.
+
-
We thought to split the <b>sense and store machineries in two different bacterial populations</b>, making them
+
-
communicate through Quorum Sensing <a href="#biblio" style="color: #8ac101">[9]</a>: we used the <i>LuxI/LuxR</i> system from <i>Alivibrio fischerii</i>. The first
+
-
population senses nickel concentration in medium ( and translates it in a fluorescence signal), then activating
+
-
<i>Acyl-Homoserine-Lactones</i> (<i>AHLs</i>) production. The second population receives the <i>QS</i> signal and sums it up to the nickel detection signal (as in an AND gate), activating <i>Hpn</i> (the storage protein) expression and <i>AHLs</i> signal amplification.
+
-
</p>
+
-
<br/>
+
-
<img src="ihttps://static.igem.org/mediawiki/2013/6/68/Unisalento_welcome2.png" alt="" width="100%">
+
-
<br/>
+
-
<br/>
+
-
</div>
+
-
<div class="clear"></div>
+
-
<!-- *************************************************************************************************** -->
+
-
<img src="images/img/dna.png" alt="" style="margin-bottom: -100px; margin-left: 360px; position: absolute">
+
-
<!-- *************************************************************************************************** -->
+
</div>
</div>
-
<!-- *************************************************************************************************** -->
+
<div class="clear"></div>
-
<div id="container">
+
<div class="nosplit">
-
<!-- *************************************************************************************************** -->
+
<p style="text-align:center;color:#cccccc">
-
<div class="nosplit">
+
<br><br>Design by Mattia Corvaglia - <a href="http://www.mattiacorvaglia.com" style="color:#cccccc">www.mattiacorvaglia.com</a> - Visitors:
-
<h2><a name="biblio">Bibliography</a></h2>
+
<!-- hitwebcounter Code START -->
-
<ol>
+
<a href="http://www.hitwebcounter.com/" target="_blank">
-
<li>Denkhaus and Salnikow, Nickel essentiality, toxicity and carcinogenity; Critical Reviews in Oncology/Hematology, 42 (2002) 35–56</li>
+
<img src="http://hitwebcounter.com/counter/counter.php?page=5095130&style=0008&nbdigits=5&type=page&initCount=0" title="URLS counter" Alt="URLS counter" border="0" >
-
<li>“Nickel, Elemental” on TOXNET- Databases on toxicology, hazardous chemicals, environmental health, and toxic releases.</li>
+
</a><br/>
-
<li>Gadd, Metals, minerals and microbes: geomicrobiology and bioremediation; Microbiology, 156 (2010), 609–643</li>
+
<!--<a href="http://www.hitwebcounter.com/" title="Free Counters" target="_blank" style="font-family: sans-serif, Arial, Helvetica; font-size: 9px; color: #9F9F97; text-decoration: none ;">
-
<li>Dosanjh and Michel, Microbial nickel metalloregulation: NikRs for nickel ions; Current Opinion in Chemical Biology, 10 (2006), 123–130</li>
+
<strong>Free Counters</strong>
-
<li>Contreras et al., Characterization of the roles of NikR, a nickel-responsive pleiotropic autoregulator of Helicobacter pylori; Molecular Microbiology, 49 (4) (2003), 947–963</li>
+
</a>-->
-
<li>Muller et al., Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori; Nucleic Acids Research, 39 (17) (2011), 7564–7575</li>
+
</p>
-
<li>Seshadri et al., Roles of His-Rich Hpn and Hpn-Like Proteins in Helicobacter pylori Nickel Physiology; Journal of Bacteriology, 189 (11) (2007), 4120–4126</li>
+
-
<li>Ge et al., Expression and characterization of a histidine-rich protein, Hpn: potential for Ni2+ storage in Helicobacter pylori; Biochemistry Journal 393 (2006), 285–293</li>
+
-
<li>Miller, Bassler, Quorum Sensing in bacteria; Annual Review of Microbiology 55 (2001), 165-99.</li>
+
-
</ol>
+
-
</div>
+
-
<!-- *************************************************************************************************** -->
+
</div>
</div>
-
<!-- *************************************************************************************************** -->
 
</div>
</div>
-
</div>
+
</footer>
-
<div class="clear"></div>
+
-
<!-- ********************* Start Footer *********************************** -->
+
-
<footer></footer>
+
<!-- ********************* End Footer ************************************* -->
<!-- ********************* End Footer ************************************* -->
<!-- ********************* Start Button "Back-to-top" *********************************** -->
<!-- ********************* Start Button "Back-to-top" *********************************** -->
<p id="back-top">
<p id="back-top">
-
<a href="#top"><span></span>Back to Top</a>
+
<a href="#top"><span></span>Back to Top</a>
</p>
</p>
<!-- ********************* End Button "Back-to-top" *********************************** -->
<!-- ********************* End Button "Back-to-top" *********************************** -->
</body>
</body>
</html>
</html>

Latest revision as of 14:43, 4 October 2013



Introduction to NickBusters Project

Nickel is one of the most widespread heavy metals in the ecosystem, and it is essential for many organisms, from bacteria to higher forms of life. However, as all the heavy metals, even a small amount of Nickel higher than the essential dose could be toxic, leading to various noxious effects [1-2]: then the need to remove its excess from many substrates. Nowadays bacteria-mediated bioremediation from inorganic substances seems to be a considerably relevant frontier in microbial biotechnologies [3]. Our project aims to develop a living system, in an easily monitorable bacterial platform, who would work as a Nickel detector and a Nickel remediation system. The whole system is based on genetic parts from the pathogen Helicobacter pylori, whose evolution led to a stronger reliance on Nickel ions for host interaction and pathogenesis. Sensing relies on nickel regulator NikR from H. pylori (HpNikR): though homolog of E.coli NikR, it has a stronger dependence on Nickel ions and a wider range of regulated elements, being a rare case of pleiotropic regulator in bacteria. As HpNikR could contemporaneously activate and repress gene expression [4-5-6], we thought to develop a synthetic gene cluster under HpNikR - regulated promoters. With a double regulation, one of the promoters being activated while the other gets repressed, the system allows to follow up Nickel sensing as a fluorescence signal from fluorescent proteins. Thus, adding bacteria to a nickel containing medium results in switching off a basal occurring green fluorescence and in switching on a red fluorescence signal.

Twitter


The Nickel storage system is even taken from our pathogenic friend: Hpn protein is a Nickel-binding protein, whose purpose is to store the Nickel ions inside the cell and to supply them with other protein interactors, from Nickel dependent heat shock proteins to pathogenesis factors as urease [7]. Hpn can be expressed in a heterologous model system (E.coli), increasing bacterial resistance up to a toxic dose of Nickel ions and allowing Ni++ intracellular accumulation [8]. We thought to split the sensing and storing machineries into two different bacterial populations, making them communicate with each other through Quorum Sensing [9]: we used the LuxI/LuxR system from Alivibrio fischerii. The first population senses nickel concentration in medium ( and translates it in a fluorescence signal), then activating Acyl-Homoserine-Lactones (AHLs) production. The second population receives the QS signal and sums it up to the nickel detection signal (as in an AND gate), activating Hpn (the storage protein) expression and AHLs signal amplification.




Bibliography
  1. Denkhaus and Salnikow, Nickel essentiality, toxicity and carcinogenity; Critical Reviews in Oncology/Hematology, 42 (2002) 35–56
  2. “Nickel, Elemental” on TOXNET- Databases on toxicology, hazardous chemicals, environmental health, and toxic releases.
  3. Gadd, Metals, minerals and microbes: geomicrobiology and bioremediation; Microbiology, 156 (2010), 609–643
  4. Dosanjh and Michel, Microbial nickel metalloregulation: NikRs for nickel ions; Current Opinion in Chemical Biology, 10 (2006), 123–130
  5. Contreras et al., Characterization of the roles of NikR, a nickel-responsive pleiotropic autoregulator of Helicobacter pylori; Molecular Microbiology, 49 (4) (2003), 947–963
  6. Muller et al., Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori; Nucleic Acids Research, 39 (17) (2011), 7564–7575
  7. Seshadri et al., Roles of His-Rich Hpn and Hpn-Like Proteins in Helicobacter pylori Nickel Physiology; Journal of Bacteriology, 189 (11) (2007), 4120–4126
  8. Ge et al., Expression and characterization of a histidine-rich protein, Hpn: potential for Ni2+ storage in Helicobacter pylori; Biochemistry Journal 393 (2006), 285–293
  9. Miller, Bassler, Quorum Sensing in bacteria; Annual Review of Microbiology 55 (2001), 165-99.

Back to Top

Retrieved from "http://2013.igem.org/Team:UniSalento_Lecce"