Team:Goettingen/Focus Groups deu

From 2012.igem.org

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<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" dir="ltr" lang="en"><head>
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{{GoettingenHeader_deu|deu=Team:Goettingen/Focus Groups_deu|eng=Team:Goettingen/Focus Groups}}
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                <meta http-equiv="Content-Type" content="text/html; charset=UTF-8">
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__TOC__
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                                <meta name="keywords" content="Team:Goettingen,Team:Goettingen,Team:Goettingen/Homing coli,Team:Goettingen/Human Practice/Flash coli,Team:Goettingen/Press,Team:Goettingen/Project/General information">
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                <link rel="shortcut icon" href="https://2012.igem.org/favicon.ico">
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                <link rel="search" type="application/opensearchdescription+xml" href="https://2012.igem.org/wiki/opensearch_desc.php" title="2012.igem.org (English)">
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                <link title="Creative Commons" type="application/rdf+xml" href="https://2012.igem.org/wiki/index.php?title=Team:Goettingen&amp;action=creativecommons" rel="meta">
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<link rel="alternate" type="application/atom+xml" title="2012.igem.org Atom Feed" href="https://2012.igem.org/wiki/index.php?title=Special:Recentchanges&amp;feed=atom">
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== Fokus Gruppe #1 - Selektion / (bakterielles) Schwimmen  ==
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<html><a href="https://static.igem.org/mediawiki/2012/4/48/Goett_taxtaskf_b.jpg"><img width="900" src="https://static.igem.org/mediawiki/2012/6/60/Goett_Team1_focus_t2.jpg"></a></html>
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<body class="mediawiki ns-0 ltr page-Team_Goettingen">
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In dieser Gruppe dreht sich alles um die Entwicklung eines Selektionssystems, welches die Auslese der am schnellsten schwimmenden <i>E. coli</i> Klone ermöglicht. In einer intensiven Literaturrecherche sammelten wir Zusammensetzungen verschiedenster Schwimmagarmedien und Anleitungen für Schwimm-Assays und evaluierten diese hinsichtlich unseres Zieles. Generell können Medien sich auf die verschiedensten Weisen voneinander unterscheiden, aber in unserem Fall sind die Agarkonzentration und der Nährstoffgehalt am bedeutendsten. Die Agarkonzentration muss niedrig genug sein, um Schwimmen zu erlauben, aber dennoch hoch genug um eine halbfeste Struktur bilden zu können.
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Da die <i>E. coli</i> Zellen nur Schwimmen, wenn sich keine Nährstoffe mehr in ihrer unmittelbaren Umgebung befinden, sollte der Nährstoffgehalt so niedrig wie möglich sein ohne dass das Wachstum der <i>E. coli</i> Zellen bedeutend beeinträchtigt wird. Der niedrige Nährstoffgehalt ist außerdem notwendig, um zu untersuchen, ob eine gerichtete Taxis in Abhängigkeit unsere getesteten Moleküle stattfindet.
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Mithilfe aller dieser unterschiedlichen Experimente werden wir in der Lage sein ein Selektionssystem zu etablieren, welches schnell und einfach zu handhaben ist und zudem reproduktive Resultate liefert.
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                        <!-- start content -->
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[[File:Goett_Coli_select.jpg|center]]
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<br><br>
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== Fokus Gruppe #2 - Verbesserung der Geschwindigkeit ==
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<style>
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<html><a href="https://static.igem.org/mediawiki/2012/9/93/Goett_team2_PS.jpg"><img width="900" src="https://static.igem.org/mediawiki/2012/0/04/Gr2_Speedsquad_t.jpg"></a></html>
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h1.firstHeading { display: none; }
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p {text-align: justify;}
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In dieser Gruppe konzentrieren wir uns auf die Verbesserung der Schwimmfähigkeiten von <i>E. coli</i>. Um dies zu erreichen wählten wir ein Set von Genen, die in das  Schwimmverhalten der E. colis involviert sein könnten. Durch die Verwendung verschiedener Promotoren können die Expressionslevel dieser Gene kontrolliert und die modifizierten Bakterien anschließend auf Schwimm-Agarplatten getestet werden.
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a:link { color: #004080; text-decoration: none}
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Zu den gewählten Genen gehören flhDC, welchesden Masterregulator für Motilität und Chemotaxis verschlüsselt oder auch motB, welches Bestandteile des bakteriellen Flagellums codiert. Weiterhin verwenden wir für unsere Experimente unterschiedliche Stämme von <i>E. coli</i> um Unterschiede in der Beweglichkeit identifizieren zu können.
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a:visited { color:##003090; text-decoration: none}
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#bodyContent { padding: 10px auto; width: 910px; margin: auto; clear: none; }
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[[File:Goett_Coli_speed.jpg]]
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<br><br>
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table#team_members { text-align: justify; border: 0; }
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== Fokus Gruppe #3 - Chemorezeptoren  ==
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table#team_members h2, table#team_members h3 { clear: both; }
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<html><a href="https://static.igem.org/mediawiki/2012/3/30/Goett_group3.jpg"><img width="900" src="https://static.igem.org/mediawiki/2012/2/21/Goett_group3_t.jpg"></a></html>
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In dieser Gruppe liegt der Hauptfokus darauf den Aspartat-Rezeptor von <i>E. coli</i> unter Kontrolle verschieden starker konstitutiver Promotoren in Vektoren zu klonieren. Nachdem die Effekte der Überexpression des Chemotaxis-Rezeptors
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/*
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untersucht wurden, ist die nächste Herausforderung die Mutagenese dieses Proteins. Diese soll der Erstellung einer Chemorezeptor-Bibliothek dienen.
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li:hover works in IE7 and FF2.
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div.MenuBar ul li:hover ul.DropDownMenu li ul.SideMenu,
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Idealerweise wird durch die Mutation der für chemisches Binden bedeutenden Liganden-Bindestelle die Spezifität des Rezeptors verändert.
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[[File:Goett_Coli_nose.jpg]]
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                  Use MenuTailor.css to customize. */
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                            Use MenuTailor.css to customize. */
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div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu,
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                  Use MenuTailor.css to customize. */
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                Keep it sync with the side menu "effective border" width. */
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  left: 13em; /* Places the side menu to the right of the drop-down menu.
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                            Keep it sync with the drop-down menu width.
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                            Use MenuTailor.css to customize. */
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div.MenuBar ul li:hover ul.DropDownMenu li:hover ul.SideMenu li a,
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                            Use MenuTailor.css to customize. */
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  font: 12px Verdana; /* Required for IE55. */
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  left: 13em; /* Places the side menu to the right of the drop-down menu.
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                            Keep it sync with the drop-down menu width.
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                            Use MenuTailor.css to customize. */
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}
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div.MenuBar ul li ul.DropDownMenu li ul.SideMenu li a span {
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  padding-left: 1.5em; /* Sets the left space of each side menu item. */
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  text-align: left;
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  font: 12px Verdana; /* Required for IE55. */
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  left: 13em; /* Places the side menu to the right of the drop-down menu.
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                            Keep it sync with the drop-down menu width.
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                            Use MenuTailor.css to customize. */
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}
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/*----------------------------------------------------------------------------- Browser Specific */
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* html div.MenuBar ul li a {
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  float: left; /* Required for IE55 and IE6.
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                            Breaks O9.
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                            Hidden (* html) from non-IE browsers. */
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}
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* html ul.DropDownMenu li a:hover {
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                  Hidden (* html) from non-IE browsers. */
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ul.DropDownMenu li a:hover {
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                      Hidding it (* html) from non-IE browsers breaks IE7!
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* html div.MenuBar a:hover {
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  text-decoration: none; /* Required for IE55 and IE6.
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                            Hidden (* html) from non-IE browsers. */
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* html div.MenuBar ul li table,
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* html div.MenuBar ul li table td {
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  border: 0; /* Required for IE55 and IE6.
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                Hidden (* html) from non-IE browsers. */
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}
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/*------------------------------------------------------------------------------- Default Colors */
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div.MenuBar {
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Sprache: <img height="20", src="http://www.patrickreinke.de/igem/eng.jpg"> <a href="https://2012.igem.org/wiki/index.php?title=Team:Goettingen/Focus_Groups">English</a>, <img height="20", src="http://www.patrickreinke.de/igem/deu.jpg"> Deutsch <br>
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<table id="toc" class="toc"><tbody><tr><td><div id="toctitle"><h2>Inhalte</h2> <span class="toctoggle">[<a href="javascript:toggleToc()" class="internal" id="togglelink">verstecken</a>]</span></div>
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<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#Focus_Groups#1"><span class="tocnumber"></span> <span class="toctext">#1 - Selektion / (bakterielles) Schwimmen </span></a></li>
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Latest revision as of 19:13, 26 September 2012

Deutsch  / English 

Contents


Fokus Gruppe #1 - Selektion / (bakterielles) Schwimmen

In dieser Gruppe dreht sich alles um die Entwicklung eines Selektionssystems, welches die Auslese der am schnellsten schwimmenden E. coli Klone ermöglicht. In einer intensiven Literaturrecherche sammelten wir Zusammensetzungen verschiedenster Schwimmagarmedien und Anleitungen für Schwimm-Assays und evaluierten diese hinsichtlich unseres Zieles. Generell können Medien sich auf die verschiedensten Weisen voneinander unterscheiden, aber in unserem Fall sind die Agarkonzentration und der Nährstoffgehalt am bedeutendsten. Die Agarkonzentration muss niedrig genug sein, um Schwimmen zu erlauben, aber dennoch hoch genug um eine halbfeste Struktur bilden zu können.

Da die E. coli Zellen nur Schwimmen, wenn sich keine Nährstoffe mehr in ihrer unmittelbaren Umgebung befinden, sollte der Nährstoffgehalt so niedrig wie möglich sein ohne dass das Wachstum der E. coli Zellen bedeutend beeinträchtigt wird. Der niedrige Nährstoffgehalt ist außerdem notwendig, um zu untersuchen, ob eine gerichtete Taxis in Abhängigkeit unsere getesteten Moleküle stattfindet.

Mithilfe aller dieser unterschiedlichen Experimente werden wir in der Lage sein ein Selektionssystem zu etablieren, welches schnell und einfach zu handhaben ist und zudem reproduktive Resultate liefert.


Goett Coli select.jpg




Fokus Gruppe #2 - Verbesserung der Geschwindigkeit

In dieser Gruppe konzentrieren wir uns auf die Verbesserung der Schwimmfähigkeiten von E. coli. Um dies zu erreichen wählten wir ein Set von Genen, die in das Schwimmverhalten der E. colis involviert sein könnten. Durch die Verwendung verschiedener Promotoren können die Expressionslevel dieser Gene kontrolliert und die modifizierten Bakterien anschließend auf Schwimm-Agarplatten getestet werden.

Zu den gewählten Genen gehören flhDC, welchesden Masterregulator für Motilität und Chemotaxis verschlüsselt oder auch motB, welches Bestandteile des bakteriellen Flagellums codiert. Weiterhin verwenden wir für unsere Experimente unterschiedliche Stämme von E. coli um Unterschiede in der Beweglichkeit identifizieren zu können.

Goett Coli speed.jpg

Fokus Gruppe #3 - Chemorezeptoren


In dieser Gruppe liegt der Hauptfokus darauf den Aspartat-Rezeptor von E. coli unter Kontrolle verschieden starker konstitutiver Promotoren in Vektoren zu klonieren. Nachdem die Effekte der Überexpression des Chemotaxis-Rezeptors untersucht wurden, ist die nächste Herausforderung die Mutagenese dieses Proteins. Diese soll der Erstellung einer Chemorezeptor-Bibliothek dienen.

Idealerweise wird durch die Mutation der für chemisches Binden bedeutenden Liganden-Bindestelle die Spezifität des Rezeptors verändert.

Goett Coli nose.jpg


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