Team:Chiba/Plasmid1

From 2010.igem.org

(Difference between revisions)
(→Reference)
 
(21 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/2010/0/09/Chiba_menu.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 56: Line 15:
       font-size:100%;
       font-size:100%;
       line-height:normal;
       line-height:normal;
-
       margin-top: -8px;
+
       margin-top: -9px;
       }
       }
     #tabsE ul {
     #tabsE ul {
         margin:0;
         margin:0;
-
         padding:2px 10px 0 20px;
+
         padding:2px 10px 0 95px;
         list-style:none;
         list-style:none;
       }
       }
Line 135: Line 94:
-->
-->
-
</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 157: Line 118:
</li>
</li>
<li><a href="https://2010.igem.org/Team:Chiba/Parts"><span>Parts</span></a></li>
<li><a href="https://2010.igem.org/Team:Chiba/Parts"><span>Parts</span></a></li>
-
<li><a href="https://2010.igem.org/Team:Chiba/Modeling"><span>Modeling</span></a></li>
 
<li><a href="https://2010.igem.org/Team:Chiba/Result"><span>Result</span></a>
<li><a href="https://2010.igem.org/Team:Chiba/Result"><span>Result</span></a>
<ul style="z-index:1"><br>
<ul style="z-index:1"><br>
Line 178: Line 138:
                                 </ul>
                                 </ul>
                         </div>
                         </div>
 +
<br>
                 </body>
                 </body>
-
<!--- Background Contents End --->
 
-
 
-
<!--- Main Contents Start --->
 
-
<br><br><br>
 
-
<font size="5">PT7/cI hybrid promoter</font>&nbsp;&nbsp;&nbsp;<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K396000">BBa_K396000</a><br>
 
-
&nbsp;&nbsp;&nbsp; low unregulated, repressed by cI, activated by T7, repression is stronger than activation.
 
</html>
</html>
 +
 +
<!-- -------------------------------------------------------------------------------
 +
-------------------------------------------------------------------------------
 +
ここから下を編集
 +
-------------------------------------------------------------------------------
 +
----------------------------------------------------------------------------------- -->
 +
 +
==T7/CI-OR1 hybrid promoter <Favorite Part>==
 +
*[http://partsregistry.org/wiki/index.php?title=Part:BBa_K396000 BBa_K396000]
 +
T7/CI-OR1 hybrid promoter was designed to be activated by T7 RNA Polymerase and repressed by lambda CI protein. In sequence design, lambda CI operator site 1 (OR1, Part of BBa_R1051) was directly attached to the downstream of T7 promoter sequence (BBa_I719005). This characterization was implemented to validate the promoter function. In results, the activation and repression was actually observed.
 +
 +
===Materials and Methods===
 +
[[Image:T7-CI-1.png|frame|center|Fig. A materials and methods]]
 +
 +
===Results and Conclusion===
 +
Under the situation of CI non-expressed , T7 RNAP causes GFP expression by activating T7/CI hybrid promoter (Fig.1). On the other hand, under the situation of CI expressed, T7 RNAP does not cause GFP expression (Fig.2). The reason of this is considered as following. Under the situation of CI expressed, even though T7 RNAP tries to activate T7/CI promoter, the promoter Is repressed by CI because the ability of CI repression is superior to the one of T7 activation for this promoter. So it is concluded that T7/CI hybrid promoter is to be activated by T7 RNA Polymerase and repressed by lambda CI protein, following the rules of CI repression > T7 activation.
 +
[[Image:CI-2.png|frame|center|Fig. B Results]]
 +
[[Image:CI-3.jpg|frame|center|Fig. C Supplementary Data]]
 +
 +
===Reference===
 +
Dubendorf, J.W. & Studier, F.W. Controlling basal expression in an
 +
inducible T7 expression system by blocking the target T7 promoter with
 +
lac repressor. Journal of Molecular Biology 219, 45-59 (1991).

Latest revision as of 16:22, 27 October 2010



Contents

T7/CI-OR1 hybrid promoter <Favorite Part>

  • [http://partsregistry.org/wiki/index.php?title=Part:BBa_K396000 BBa_K396000]

T7/CI-OR1 hybrid promoter was designed to be activated by T7 RNA Polymerase and repressed by lambda CI protein. In sequence design, lambda CI operator site 1 (OR1, Part of BBa_R1051) was directly attached to the downstream of T7 promoter sequence (BBa_I719005). This characterization was implemented to validate the promoter function. In results, the activation and repression was actually observed.

Materials and Methods

Fig. A materials and methods

Results and Conclusion

Under the situation of CI non-expressed , T7 RNAP causes GFP expression by activating T7/CI hybrid promoter (Fig.1). On the other hand, under the situation of CI expressed, T7 RNAP does not cause GFP expression (Fig.2). The reason of this is considered as following. Under the situation of CI expressed, even though T7 RNAP tries to activate T7/CI promoter, the promoter Is repressed by CI because the ability of CI repression is superior to the one of T7 activation for this promoter. So it is concluded that T7/CI hybrid promoter is to be activated by T7 RNA Polymerase and repressed by lambda CI protein, following the rules of CI repression > T7 activation.

Fig. B Results
Fig. C Supplementary Data

Reference

Dubendorf, J.W. & Studier, F.W. Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor. Journal of Molecular Biology 219, 45-59 (1991).

Retrieved from "http://2010.igem.org/Team:Chiba/Plasmid1"