Team:Tsinghua-A/Parts

From 2011.igem.org

(Difference between revisions)
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1147</SPAN></FONT></FONT></P>
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1147</SPAN></FONT></FONT></P>
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</TD>
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</TR>
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</TABLE>
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<BR>
 +
<h1>Descriptions</h1><hr width="100%" size=2 color=gray>
 +
<br>
 +
<p>We assembled <A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8">BBa_E0840</FONT></A>
 +
under the pLas promoter
 +
<A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_R0079"><FONT COLOR="#002bb8">BBa_R0079</FONT></A>
 +
that was contained into K574009. We kept pSB1A2 as the scaffold vector.</p>
 +
<br>
 +
<h1>Methods</h1><hr width="100%" size=2 color=gray>
 +
<br>
 +
<p>1. Ligate
 +
<A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8">BBa_E0840</FONT></A>
 +
downstream of
 +
<A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_K574009"><FONT COLOR="#002bb8">BBa_K574009</FONT></A>
 +
, which acts as a reporter.</p>
 +
 +
<p>2. Culture the positive colony at 37&deg;C,
 +
220 rpm for 12 hours, as well as the DH5alpha.</p>
 +
 +
<p>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.</p>
 +
 +
<p>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.</p>
 +
 +
<p>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).</p>
 +
 +
<P>6. Measure the fluorescence of the samples with the flow cytometer.</P>
 +
<br>
 +
<h1>Results</h1><hr width="100%" size=2 color=gray>
 +
<br>
 +
<p style="text-indent:0em"align="CENTER"><img src="https://static.igem.org/mediawiki/2011/c/ca/K574009_chart.png" NAME="K574009_chart" /></p>
 +
<P>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<sup>-5</sup>M, and higher concentration may lead to the
 +
expression of GFP decreasing. Additionally, to most groups, the intensity of GFP
 +
reached its maxium after 4 hours.</P>
 +
 +
 +
<br><br><br>
 +
<p align="CENTER" STYLE="text-indent:0em"><a href="https://2011.igem.org/Team:Tsinghua-A"><img src="https://static.igem.org/mediawiki/2011/9/92/Killbanner_header.jpg" alt="" width="960"/><a href="https://2011.igem.org"><img src="https://static.igem.org/mediawiki/igem.org/2/29/Killbanner_header2.jpg" alt="" width="960"/></p>
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</BODY>
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</HTML>
 +
{{:Team:Tsinghua-A/template/killbanner}}
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<html xmlns="http://www.w3.org/1999/xhtml">
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<head>
 +
<meta http-equiv="Content-Type" content="text/html; charset=iso-8859-1" />
 +
<title>JavaScript Slideshow - TinySlideshow</title>
 +
<link rel="stylesheet" href="http://bioinfo.au.tsinghua.edu.cn/member/IGEM/thu_style.css" />
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<style>
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 +
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body {
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margin: 0;
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padding: 0;
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 +
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 +
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 +
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 +
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 +
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 +
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 +
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 +
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 +
 +
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 +
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 +
border-left: 1px solid #FFFFFF;
 +
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 +
 +
#menu a {
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display: block;
 +
width: 155px;
 +
height: 33px;
 +
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 +
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 +
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 +
 +
#menu a:hover, #menu .current_page_item a {
 +
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 +
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 +
 +
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#menu .normal_item a {
 +
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 +
</style>
 +
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</head>
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<body>
 +
<p 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">
 +
<ul>
 +
<li><a href="https://2011.igem.org/Team:Tsinghua-A/Team">Team</a></li>
 +
<li><a href="https://2011.igem.org/Team:Tsinghua-A/Project">Project</a></li>
 +
<li><a href="https://2011.igem.org/Team:Tsinghua-A/Safety">Safety</a></li>
 +
<li ><a href="https://2011.igem.org/Team:Tsinghua-A/Modeling">Modeling</a></li>
 +
<li class="current_page_item"><a href="https://2011.igem.org/Team:Tsinghua-A/Parts">Parts</a></li>
 +
<li ><a href="https://2011.igem.org/Team:Tsinghua-A/Notebook">Notebook</a></li>
 +
</ul>
 +
</div>
 +
 +
<!--end of enu--------------------------------------------><!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.0 Transitional//EN">
 +
<HTML>
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<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="20111002;3132500">
 +
<META NAME="CHANGEDBY" CONTENT="Sl Y">
 +
<META NAME="CHANGED" CONTENT="20111002;20522400">
 +
<META NAME="CHANGEDBY" CONTENT="Sl Y">
 +
</HEAD>
 +
<BODY LANG="zh-CN" DIR="LTR">
 +
<br>
 +
 +
<h1>Parts List</h1><hr width="100%" size=2 color=gray>
 +
<br>
 +
<TABLE Align=CENTER WIDTH=760 BORDER=5 CELLPADDING=0 CELLSPACING=0 BGCOLOR="#ccccff">
 +
<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 text-indent:0em text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Name</SPAN></FONT></FONT></P>
 +
</TH>
 +
<TH WIDTH=130 BGCOLOR="#aaaaff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Type</SPAN></FONT></FONT></P>
 +
</TH>
 +
<TH WIDTH=340 BGCOLOR="#aaaaff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Description</SPAN></FONT></FONT></P>
 +
</TH>
 +
<TH WIDTH=130 BGCOLOR="#aaaaff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Length</SPAN></FONT></FONT></P>
 +
</TH>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pLux + TetR</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">903</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">TetR
 +
regulated by 3OC6HSL</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1890</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pluxR
 +
+ CFP</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">941</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC6HSL -> TetR && reporter</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2839</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL regulated by TetR</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">797</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL and YFP regulated by pBad</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1712</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL regulated by pTet and pBad</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2517</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">lasR + TT</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">918</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">a constitutive LasR producer</SPAN></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">982</SPAN></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><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 text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL -> PoPS Receiver</SPAN></FONT></FONT></P>
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<TD WIDTH=130 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1147</SPAN></FONT></FONT></P>
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Revision as of 08:23, 26 October 2011

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Parts List


Name

Type

Description

Length

BBa_K574000

Composite

pLux + TetR

903

BBa_K574001

Composite

TetR regulated by 3OC6HSL

1890

BBa_K574002

Composite

pluxR + CFP

941

BBa_K574003

Device

3OC6HSL -> TetR && reporter

2839

BBa_K574004

Composite

3OC12HSL regulated by TetR

797

BBa_K574005

Composite

3OC12HSL and YFP regulated by pBad

1712

BBa_K574006

Device

3OC12HSL regulated by pTet and pBad

2517

BBa_K574007

Composite

lasR + TT

918

BBa_K574008

Composite

a constitutive LasR producer

982

BBa_K574009

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 maxium after 4 hours.




JavaScript Slideshow - TinySlideshow


Parts List


Name

Type

Description

Length

BBa_K574000

Composite

pLux + TetR

903

BBa_K574001

Composite

TetR regulated by 3OC6HSL

1890

BBa_K574002

Composite

pluxR + CFP

941

BBa_K574003

Device

3OC6HSL -> TetR && reporter

2839

BBa_K574004

Composite

3OC12HSL regulated by TetR

797

BBa_K574005

Composite

3OC12HSL and YFP regulated by pBad

1712

BBa_K574006

Device

3OC12HSL regulated by pTet and pBad

2517

BBa_K574007

Composite

lasR + TT

918

BBa_K574008

Composite

a constitutive LasR producer

982

BBa_K574009

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 maxium after 4 hours.




JavaScript Slideshow - TinySlideshow


Parts List


Name

Type

Description

Length

BBa_K574000

Composite

pLux + TetR

903

BBa_K574001

Composite

TetR regulated by 3OC6HSL

1890

BBa_K574002

Composite

pluxR + CFP

941

BBa_K574003

Device

3OC6HSL -> TetR && reporter

2839

BBa_K574004

Composite

3OC12HSL regulated by TetR

797

BBa_K574005

Composite

3OC12HSL and YFP regulated by pBad

1712

BBa_K574006

Device

3OC12HSL regulated by pTet and pBad

2517

BBa_K574007

Composite

lasR + TT

918

BBa_K574008

Composite

a constitutive LasR producer

982

BBa_K574009

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 maxium after 4 hours.




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