"
Team:Tsinghua-A/Parts
From 2011.igem.org
(Difference between revisions)
| (13 intermediate revisions not shown) | |||
| Line 35: | Line 35: | ||
color:#548dd4; | color:#548dd4; | ||
font-weight:bold; | font-weight:bold; | ||
| - | font-size: | + | font-size:130%; |
padding:0; | padding:0; | ||
border:none; | border:none; | ||
margin:0; | margin:0; | ||
| + | } | ||
| + | h2{ | ||
| + | line-height:2; | ||
| + | font-family:"Times New Roman",Georgia,Serif; | ||
| + | color:#005555; | ||
| + | font-size:120%; | ||
} | } | ||
/* Menu */ | /* Menu */ | ||
| Line 94: | Line 100: | ||
</head> | </head> | ||
<body> | <body> | ||
| - | <p align="CENTER"><img src="https://static.igem.org/mediawiki/2011/1/17/THU_banner_par.jpg" alt="" width="960"/></p> | + | <div style="position:fixed;right:0px;bottom: 0px;display:inline"><a href="#back-top"> |
| + | <img src="https://static.igem.org/mediawiki/2011/5/52/Back-top.png" width="60px" height="60px"></a></div> | ||
| + | <p id="back-top" align="CENTER" STYLE="text-indent:0em"><img src="https://static.igem.org/mediawiki/2011/1/17/THU_banner_par.jpg" alt="" width="960"/></p> | ||
<div id="menu"> | <div id="menu"> | ||
<ul> | <ul> | ||
| Line 119: | Line 127: | ||
</HEAD> | </HEAD> | ||
<BODY LANG="zh-CN" DIR="LTR"> | <BODY LANG="zh-CN" DIR="LTR"> | ||
| - | <br> | + | <br><br> |
| - | + | <div class="nav"> | |
| - | < | + | <table id="toc" class="toc"> |
| - | + | <tr> | |
| - | < | + | <td> |
| - | + | <div id="toctitle"> | |
| - | + | <h2>Contents</h2> | |
| - | + | </div> | |
| - | + | <ul> | |
| - | + | <li class="toclevel-1 tocsection-1"><a href="#Parts List"><span class="tocnumber">1</span> <span class="toctext">Parts List</span></a></li> | |
| - | + | <li class="toclevel-1 tocsection-2"><a href="#Descriptions"><span class="tocnumber">2</span> <span class="toctext">Descriptions</span></a></li> | |
| - | + | <li class="toclevel-1 tocsection-3"><a href="#Methods"><span class="tocnumber">3</span> <span class="toctext">Methods</span></a></li> | |
| - | + | <li class="toclevel-1 tocsection-4"><a href="#Results"><span class="tocnumber">4</span> <span class="toctext">Results</span></a></li> | |
| - | + | </ul> | |
| - | + | </td> | |
| - | + | </tr> | |
| - | + | </table> | |
| - | + | </div> | |
| - | + | <br><!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.0 Transitional//EN"> | |
| - | + | <HTML> | |
| - | + | <HEAD> | |
| - | + | <META HTTP-EQUIV="CONTENT-TYPE" CONTENT="text/html; charset=gb2312"> | |
| - | + | <TITLE></TITLE> | |
| - | + | <META NAME="GENERATOR" CONTENT="OpenOffice.org 3.2 (Win32)"> | |
| - | + | <META NAME="AUTHOR" CONTENT="Sl Y"> | |
| - | + | <META NAME="CREATED" CONTENT="20111028;10190100"> | |
| - | + | <META NAME="CHANGEDBY" CONTENT="Sl Y"> | |
| - | + | <META NAME="CHANGED" CONTENT="20111028;10224900"> | |
| - | + | </HEAD> | |
| - | + | <BODY LANG="zh-CN" DIR="LTR"> | |
| - | + | <H1 id="Parts List"><A NAME="Parts List"></A><FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US"><B>Parts | |
| - | + | List</B></SPAN></FONT></FONT></FONT></H1> | |
| - | + | <HR SIZE=1> | |
| - | + | <P STYLE="margin-bottom: 0cm; line-height: 0.58cm; widows: 2; orphans: 2"> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | < | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | < | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
<BR> | <BR> | ||
| - | < | + | </P> |
| - | + | <CENTER> | |
| - | < | + | <TABLE WIDTH=760 BORDER=4 CELLPADDING=0 CELLSPACING=0 BGCOLOR="#ffffff"> |
| - | + | <COL WIDTH=160> | |
| - | + | <COL WIDTH=130> | |
| - | + | <COL WIDTH=340> | |
| - | + | <COL WIDTH=130> | |
| - | < | + | <TR> |
| - | + | <TH WIDTH=160 BGCOLOR="#aaaaff"> | |
| - | < | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Name</SPAN></FONT></FONT></FONT></P> |
| - | <A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8">BBa_E0840</FONT></A> | + | </TH> |
| - | downstream | + | <TH WIDTH=130 BGCOLOR="#aaaaff"> |
| - | <A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_K574009"><FONT COLOR="#002bb8">BBa_K574009</FONT></A> | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Type</SPAN></FONT></FONT></FONT></P> |
| - | , which acts as a reporter.</ | + | </TH> |
| - | + | <TH WIDTH=340 BGCOLOR="#aaaaff"> | |
| - | < | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Description</SPAN></FONT></FONT></FONT></P> |
| - | 220 rpm for 12 hours, as well as the DH5alpha.</ | + | </TH> |
| - | + | <TH WIDTH=130 BGCOLOR="#aaaaff"> | |
| - | < | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Length</SPAN></FONT></FONT></FONT></P> |
| - | each 3 tubes as a group. The later is also diluted in 3 tubes as | + | </TH> |
| - | the negative control.</ | + | </TR> |
| - | + | <TR> | |
| - | < | + | <TD WIDTH=160 BGCOLOR="#ddddff"> |
| - | with 3OC12HSL 0, 1e-7 M, 1e-6 M, 1e-5 M and 1e-4 M respectively.</ | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574000"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574000</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> |
| - | + | </TD> | |
| - | < | + | <TD WIDTH=130 BGCOLOR="#ddddff"> |
| - | centrifugalizate 0.5 mL of each culture and suspend them by 1 mL of | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> |
| - | (phosphate-buffered saline).</ | + | </TD> |
| - | + | <TD WIDTH=340 BGCOLOR="#ddddff"> | |
| - | <P>6. Measure the fluorescence of the samples with the flow cytometer.</P> | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pLux |
| - | + | + TetR</SPAN></FONT></FONT></FONT></P> | |
| - | < | + | </TD> |
| - | < | + | <TD WIDTH=130 BGCOLOR="#ddddff"> |
| - | <P>From | + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">903</SPAN></FONT></FONT></FONT></P> |
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574001"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574001</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">TetR | ||
| + | regulated by 3OC6HSL</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1890</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574002"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574002</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pluxR | ||
| + | + CFP</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">941</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574003"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574003</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC6HSL | ||
| + | -> TetR && reporter</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2839</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574004"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574004</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL | ||
| + | regulated by TetR</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">797</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574005"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574005</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL | ||
| + | and YFP regulated by pBad</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1712</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574006"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574006</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL | ||
| + | regulated by pTet and pBad</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2517</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574007"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574007</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">lasR | ||
| + | + TT</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">918</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574008"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574008</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">a | ||
| + | constitutive LasR producer</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">982</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | <TR> | ||
| + | <TD WIDTH=160 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574009"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574009</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=340 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL | ||
| + | -> PoPS Receiver</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | <TD WIDTH=130 BGCOLOR="#ddddff"> | ||
| + | <P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1147</SPAN></FONT></FONT></FONT></P> | ||
| + | </TD> | ||
| + | </TR> | ||
| + | </TABLE> | ||
| + | </CENTER> | ||
| + | <P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <H1 id="Descriptions" STYLE="page-break-before: always; page-break-after: avoid"><A NAME="Descriptions"></A> | ||
| + | <FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US">Descriptions</SPAN></FONT></FONT></FONT></H1> | ||
| + | <HR SIZE=1> | ||
| + | <P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <P STYLE="margin-left: 1.13cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">We | ||
| + | assembled </SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_E0840</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"> under | ||
| + | the pLas promoter </SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_R0079"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_R0079</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"> that | ||
| + | was contained into K574009. We kept pSB1A2 as the scaffold vector.</SPAN></SPAN></FONT></FONT></FONT></SPAN></FONT></P> | ||
| + | <P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <H1 id="Methods" STYLE="page-break-before: always; page-break-after: avoid"><A NAME="Methods"></A> | ||
| + | <FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US">Methods</SPAN></FONT></FONT></FONT></H1> | ||
| + | <HR SIZE=1> | ||
| + | <P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">1. | ||
| + | Ligate </SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_E0840</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"> downstream | ||
| + | of </SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_K574009"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_K574009</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal"> , | ||
| + | which acts as a reporter.</SPAN></SPAN></FONT></FONT></FONT></SPAN></FONT></P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">2. | ||
| + | Culture the positive colony at 37°C, 220 rpm for 12 hours, as | ||
| + | well as the DH5alpha.</SPAN></FONT></FONT></FONT></P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">3. | ||
| + | Dilute 1:25 the former in 15 falcon tubes (4 ml cultures), each 3 | ||
| + | tubes as a group. The later is also diluted in 3 tubes as the | ||
| + | negative control.</SPAN></FONT></FONT></FONT></P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">4. | ||
| + | After culturing for another 2 hours, as their OD600 reach 0.2, induce | ||
| + | with 3OC12HSL 0, 1e-7 M, 1e-6 M, 1e-5 M and 1e-4 M respectively.</SPAN></FONT></FONT></FONT></P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">5. | ||
| + | Take samples after 1 hour, 4 hours, 8 hours and 24 hours, that is, | ||
| + | centrifugalizate 0.5 mL of each culture and suspend them by 1 mL of | ||
| + | PBS (phosphate-buffered saline).</SPAN></FONT></FONT></FONT></P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">6. | ||
| + | Measure the fluorescence of the samples with the flow cytometer.</SPAN></FONT></FONT></FONT></P> | ||
| + | <P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <H1 id="Results" STYLE="page-break-before: always; page-break-after: avoid"><A NAME="Results"></A> | ||
| + | <FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US">Results</SPAN></FONT></FONT></FONT></H1> | ||
| + | <HR SIZE=1> | ||
| + | <P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <P ALIGN=CENTER STYLE="line-height: 200%; widows: 2; orphans: 2"><FONT COLOR="#000000"><IMG SRC="https://static.igem.org/mediawiki/2011/c/ca/K574009_chart.png" NAME="K574009_chart" ALIGN=BOTTOM WIDTH=770 HEIGHT=490 BORDER=0></FONT></P> | ||
| + | <P ALIGN=CENTER STYLE="line-height: 200%; widows: 2; orphans: 2"><BR><BR> | ||
| + | </P> | ||
| + | <P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2"> | ||
| + | <FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">From | ||
the chart, we can see that cells were hardly induced in the control | the chart, we can see that cells were hardly induced in the control | ||
group, and with the concentration of inducer growing, the intensity | group, and with the concentration of inducer growing, the intensity | ||
of GFP increased by groups. The most efficient concentration of | of GFP increased by groups. The most efficient concentration of | ||
| - | inducer was around 10< | + | inducer was around 10<SUP>-5</SUP>M, and higher concentration may |
| - | + | lead to the expression of GFP decreasing. Additionally, to most | |
| - | reached its | + | groups, the intensity of GFP reached its maximum after 4 hours.</SPAN></FONT></FONT></FONT></P> |
| - | + | <P><FONT FACE="Arial, sans-serif"><SPAN LANG="en-US"><BR></SPAN></FONT><BR><BR> | |
| - | + | </P> | |
| - | < | + | |
| - | < | + | |
</BODY> | </BODY> | ||
</HTML> | </HTML> | ||
Latest revision as of 02:28, 28 October 2011
Parts List
|
Name |
Type |
Description |
Length |
|---|---|---|---|
|
Composite |
pLux + TetR |
903 |
|
|
Composite |
TetR regulated by 3OC6HSL |
1890 |
|
|
Composite |
pluxR + CFP |
941 |
|
|
Device |
3OC6HSL -> TetR && reporter |
2839 |
|
|
Composite |
3OC12HSL regulated by TetR |
797 |
|
|
Composite |
3OC12HSL and YFP regulated by pBad |
1712 |
|
|
Device |
3OC12HSL regulated by pTet and pBad |
2517 |
|
|
Composite |
lasR + TT |
918 |
|
|
Composite |
a constitutive LasR producer |
982 |
|
|
Composite |
3OC12HSL -> PoPS Receiver |
1147 |
Descriptions
We assembled BBa_E0840 under the pLas promoter BBa_R0079 that was contained into K574009. We kept pSB1A2 as the scaffold vector.
Methods
1. Ligate BBa_E0840 downstream of BBa_K574009 , which acts as a reporter.
2. Culture the positive colony at 37°C, 220 rpm for 12 hours, as well as the DH5alpha.
3. Dilute 1:25 the former in 15 falcon tubes (4 ml cultures), each 3 tubes as a group. The later is also diluted in 3 tubes as the negative control.
4. After culturing for another 2 hours, as their OD600 reach 0.2, induce with 3OC12HSL 0, 1e-7 M, 1e-6 M, 1e-5 M and 1e-4 M respectively.
5. Take samples after 1 hour, 4 hours, 8 hours and 24 hours, that is, centrifugalizate 0.5 mL of each culture and suspend them by 1 mL of PBS (phosphate-buffered saline).
6. Measure the fluorescence of the samples with the flow cytometer.
Results
From the chart, we can see that cells were hardly induced in the control group, and with the concentration of inducer growing, the intensity of GFP increased by groups. The most efficient concentration of inducer was around 10-5M, and higher concentration may lead to the expression of GFP decreasing. Additionally, to most groups, the intensity of GFP reached its maximum after 4 hours.
