Team:Chiba/lux promoter

From 2010.igem.org

(Difference between revisions)
(→Results)
 
(88 intermediate revisions not shown)
Line 1: Line 1:
 +
<!-- ---------------------------------------------------------
 +
--------------------------------------------------------------
 +
下の方に「ここから下を編集」というところがあります
 +
そっちを編集してください
 +
--------------------------------------------------------------
 +
------------------------------------------------------------ -->
 +
 +
<html>
<html>
<style type="text/css">
<style type="text/css">
-
#top-section {
 
-
    height: 0px;
 
-
    margin-top: -80px;
 
-
    margin-left: auto;
 
-
    margin-right: auto;
 
-
    margin-bottom: 0 !important;
 
-
    margin-bottom: 0px;
 
-
    padding:0;
 
-
    border: 0;
 
-
    }
 
-
</style>
 
-
</html>
 
-
<!--- Background Contents --->
 
-
<html><br><br><br>
 
-
<div id="search-controls">
 
-
<form action="/Special:Search" id="searchform">
 
-
<input id="searchInput" name="search" type="text" title="Search 2010.igem.org [f]" accesskey="f" value="" />
 
-
<input type='submit' name="go" class="searchButton" id="searchGoButton" value="Go" title="Go to a page with this exact name if exists" />&nbsp;
 
-
      <input type='submit' name="fulltext" class="searchButton" id="mw-searchButton" value="Search" title="Search the pages for this text" />
 
-
</form>
 
-
</div> <!-- close search controls -->
 
-
 
-
 
-
<body bgcolor="white" text="black" link="yellow" vlink="purple" alink="red" background="https://static.igem.org/mediawiki/2010/a/a1/Chiba_ground_2.jpg">
 
-
<img src="https://static.igem.org/mediawiki/2010/f/f9/Chiba_menu_2.jpg">
 
-
<p>
 
-
<style type="text/css">
 
-
<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
 
-
        <html xmlns="http://www.w3.org/1999/xhtml">
 
-
                <head>
 
-
                        <meta http-equiv="Content-Type" content="text/html; charset=iso-8859-1" />
 
-
                        <title>Free CSS Navigation Menu Designs 1 at exploding-boy.com</title>
 
-
                        <style type="text/css">
 
-
<!--
 
-
    body {
 
-
        margin:0;
 
-
        padding:0;
 
-
        font: bold 11px/1.5em Verdana;
 
-
}
 
-
 
-
h2 {
 
-
        font: bold 14px Verdana, Arial, Helvetica, sans-serif;
 
-
        color: #000;
 
-
        margin: 0px;
 
-
        padding: 0px 0px 0px 15px;
 
-
}
 
-
 
/*- Menu Tabs E--------------------------- */
/*- Menu Tabs E--------------------------- */
Line 135: Line 96:
-->
-->
-
</style>
 
-
                </head>
 
-
                <body>
+
 
-
                        <div id="tabsE">
+
 
 +
</style>
 +
<body bgcolor="white" text="black" link="yellow" vlink="purple" alink="red" background="https://static.igem.org/mediawiki/2010/a/a1/Chiba_ground_2.jpg">
 +
<img src="https://static.igem.org/mediawiki/2010/f/f9/Chiba_menu_2.jpg">
 +
<div id="tabsE">
                                 <ul>
                                 <ul>
                                         <!-- CSS Tabs -->
                                         <!-- CSS Tabs -->
Line 177: Line 140:
                                 </ul>
                                 </ul>
                         </div>
                         </div>
 +
<br>
 +
                </body>
</html>
</html>
-
<!--- Background Contents End --->
 
-
<!--- Main Contents Start --->
 
-
<br><br><br>
 
-
<font size="5">Lux promoter inverter</font>&nbsp;&nbsp;&nbsp;<br><br>[[Image:Plux inv-GFP.png|thumb|Fig. 1 cloning process]]
 
-
We've charaterized Plux inverter.<br>
+
<!-- -------------------------------------------------------------------------------
-
There is promoter (HSL-luxR repressor), biobrick number R0061.<br>  
+
-------------------------------------------------------------------------------
-
We've made Plux inv-GFP and characterized about it.<br>
+
ここから下を編集
-
[[Image:lux inverter function.png|thumb|Fig. 2  function check]]
+
-------------------------------------------------------------------------------
 +
----------------------------------------------------------------------------------- -->
 +
 
 +
==Abstract==
 +
 
 +
LuxR is an AHL-dependent activator.LuxR-AHL complex bind lux box,a 20-bp sequence and activate transcription.  However the lux box is inserted between -35 and -10 and LuxR functions as LuxR-AHL inverter (Plux inv) .The Plux inv was resistered in Biobrick number R0061.We've constructed Plux inv-GFP combinig R0061and E0240 and characterized about it.<br>
 +
 
 +
==Experiments==
 +
===1.Preparing Plux inv-GFP===
 +
 
 +
Plasmid:R0061 and E0240<br>
 +
Strain:XL10-G<br>
 +
protocol<br>
 +
Preparing process of Plux inv-GFP  is shown in '''Fig. 1'''.We transformed preparing Plux inv-GFP in XL10-G and confirmed the expression GFP Fluorescence.The sequence is also conformed.
 +
 
 +
[[Image:Plux inv-GFP.png|none|frame|right|'''Fig. 1''' 　Evaluation of LuxR Inverter]]
 +
 
 +
===2.Characterization Plux inv-GFP===
 +
 
 +
Plasmid:Plux inv-GFP and Plac-LuxR (LuxR constitutive generator)<br>
 +
strain:DH10B<br>
 +
 
 +
protocol<br>
 +
We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B.
 +
1,LBL
 +
incubation for12 h at 37゜C.
 +
 
 +
==Results==
 +
 
 +
We confirmed GFP Fluorescence both AHL+ and -sample.<br>
 +
<br>
 +
[[Image:Plux inv results.png|frame|none|Fig. 3 ]]
 +
 
 +
==discussion==
 +
私たちの系では，LuxRの抑制を確認することができなかった。
 +
 
 +
==Reference==
 +
#Egland.K.A, and Greenberg.E.P, Conversion of the ''Vibrio Fischeri'' Transcriptional Activator,LuxR, to a Repressor, ''J. Bacteriol.'', '''182''', P.805-811 (2000)
 +
#Cox.R.S.3rd, Surette.M.G, Elowitz.M.B, Programming gene expression with combinatorial promoters, ''Mol Syst Biol'', '''3''' ,145 (2007)

Latest revision as of 13:29, 24 January 2011




Contents

Abstract

LuxR is an AHL-dependent activator.LuxR-AHL complex bind lux box,a 20-bp sequence and activate transcription. However the lux box is inserted between -35 and -10 and LuxR functions as LuxR-AHL inverter (Plux inv) .The Plux inv was resistered in Biobrick number R0061.We've constructed Plux inv-GFP combinig R0061and E0240 and characterized about it.

Experiments

1.Preparing Plux inv-GFP

Plasmid:R0061 and E0240
Strain:XL10-G
protocol
Preparing process of Plux inv-GFP is shown in Fig. 1.We transformed preparing Plux inv-GFP in XL10-G and confirmed the expression GFP Fluorescence.The sequence is also conformed.

Fig. 1 　Evaluation of LuxR Inverter

2.Characterization Plux inv-GFP

Plasmid:Plux inv-GFP and Plac-LuxR (LuxR constitutive generator)
strain:DH10B

protocol
We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B. 1,LBL incubation for12 h at 37゜C.

Results

We confirmed GFP Fluorescence both AHL+ and -sample.

Fig. 3

discussion

私たちの系では，LuxRの抑制を確認することができなかった。

Reference

  1. Egland.K.A, and Greenberg.E.P, Conversion of the Vibrio Fischeri Transcriptional Activator,LuxR, to a Repressor, J. Bacteriol., 182, P.805-811 (2000)
  2. Cox.R.S.3rd, Surette.M.G, Elowitz.M.B, Programming gene expression with combinatorial promoters, Mol Syst Biol, 3 ,145 (2007)
Retrieved from "http://2010.igem.org/Team:Chiba/lux_promoter"