Team:Cornell/project/wetlab/chassis

From 2014.igem.org

(Difference between revisions)
 
(7 intermediate revisions not shown)
Line 7: Line 7:
});
});
</script>
</script>
-
<div class="container" id="top">
+
<body>
 +
<div class="container">
<div class="row">
<div class="row">
<div class="col-md-9 col-xs-18">
<div class="col-md-9 col-xs-18">
-
<h1 style="margin-top: 0px;"><i>E. coli DH5α</i></h1>
+
<h1><i>E. coli</i> DH5α</h1>  
-
<i>E. coli</i> is a gram-negative, facultative anaerobic, rod-shaped bacterium that is the most widely studied prokaryotic model organism for work with recombinant DNA. We are using electrocompotent and chemically compotent DH5α strains that contain recA1 and endA1 mutations that increase insert stability and improve the quality of plasmid DNA prepared from minipreps <sup>[1]</sup>.
+
<i>E. coli</i> is a gram-negative, facultative anaerobic, rod-shaped bacterium that is the most widely studied prokaryotic model organism for work with recombinant DNA. We are using electrically competent and chemically competent DH5α strains that contain <i>recA1</i> and <i>endA1</i> mutations that increase insert stability and improve the quality of plasmid DNA prepared from minipreps.<sup>[1]</sup>
-
<br><br>
+
                <h1><i>E. coli</i> BL21</h1>
-
                                <h1><i>E. coli BL21</i></h1>
+
We are also using BL21 competent strains of <i>E. coli</i>, which are designed for high-efficiency protein expression of any gene under control of the T7 bacteriophage promoter system. T7 RNA Polymerase is regulated by arabinose induction and glucose inhibition of the <i>araBAD</i> promoter in its chromosomal DNA. Proteins under control of <i>T7</i> promoter-driven vectors are then expressed at high levels. The cells also contain the T7 lysozyme gene within the pLysS plasmid, which suppresses T7 RNA polymerase activity caused by leaky basal expression.<sup>[2]</sup>
-
                                We are also using BL21 competent strains of <i>E. coli</i>, which are designed for high-efficiency protein expression of any gene under control of the T7 bacteriophage promoter system. T7 RNA Polymerase is regulated by arabinose induction and glucose inhibition of the <i>ara</i>BAD promoter in its chromosomal DNA. Proteins under control of T7 promoter driven vectors are then expressed at high levels. The cells also contain the T7 lysozyme gene within the pLysS plasmid, which suppresses T7 RNA polymerase activity caused by leaky basal expression <sup>[2]</sup>.
+
</div>
</div>
<div class="col-md-3 col-xs-5">
<div class="col-md-3 col-xs-5">
-
<a href="https://static.igem.org/mediawiki/2014/5/5a/Cornell_ecoli.png" data-toggle="lightbox" data-gallery="metal" class="thumbnail">
+
<div class="thumbnail">
<img src="https://static.igem.org/mediawiki/2014/5/5a/Cornell_ecoli.png">
<img src="https://static.igem.org/mediawiki/2014/5/5a/Cornell_ecoli.png">
-
</a>
 
</div>
</div>
</div>
</div>
</div>
</div>
-
+
<div class="row">
 +
<div class="col-md-12">
<h1>References</h1>
<h1>References</h1>
<hr>
<hr>
<ol>
<ol>
<li>Dh5-Alpha Competent <i>E. coli</i>. (2013). Retrieved from: http://www.mclab.com/Dh5-Alpha-Competent-E.-Coli.html </li>
<li>Dh5-Alpha Competent <i>E. coli</i>. (2013). Retrieved from: http://www.mclab.com/Dh5-Alpha-Competent-E.-Coli.html </li>
-
<li>Competent Cell BL21(DE3)pLysS. (2013). Retrieved from: http://www.biodynamics.co.jp/images/prd_ds250/DS260BLysShp.pdf  
+
<li>Competent Cell BL21(DE3)pLysS. (2013). Retrieved from: http://www.biodynamics.co.jp/images/prd_ds250/DS260BLysShp.pdf</li>
-
</li>
+
</ol>
-
<br><br>
+
</div>
</div>
</div>
</div>
</div>
</div>
-
<span id="top-link-block" class="hidden">
 
-
<a href="#top" class="well well-sm" onclick="$('html,body').animate({scrollTop:0},'slow');return false;">
 
-
<i class="glyphicon glyphicon-chevron-up"></i> Back to Top
 
-
</a>
 
-
</span>
 
   </body>
   </body>
</html>
</html>

Latest revision as of 03:37, 18 October 2014

Cornell iGEM

web stats

Wet Lab

E. coli DH5α

E. coli is a gram-negative, facultative anaerobic, rod-shaped bacterium that is the most widely studied prokaryotic model organism for work with recombinant DNA. We are using electrically competent and chemically competent DH5α strains that contain recA1 and endA1 mutations that increase insert stability and improve the quality of plasmid DNA prepared from minipreps.[1]

E. coli BL21

We are also using BL21 competent strains of E. coli, which are designed for high-efficiency protein expression of any gene under control of the T7 bacteriophage promoter system. T7 RNA Polymerase is regulated by arabinose induction and glucose inhibition of the araBAD promoter in its chromosomal DNA. Proteins under control of T7 promoter-driven vectors are then expressed at high levels. The cells also contain the T7 lysozyme gene within the pLysS plasmid, which suppresses T7 RNA polymerase activity caused by leaky basal expression.[2]

References


  1. Dh5-Alpha Competent E. coli. (2013). Retrieved from: http://www.mclab.com/Dh5-Alpha-Competent-E.-Coli.html
  2. Competent Cell BL21(DE3)pLysS. (2013). Retrieved from: http://www.biodynamics.co.jp/images/prd_ds250/DS260BLysShp.pdf