Difference between revisions of "Team:Lethbridge/Improve"

 
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{{Team:Lethbridge/assets/reset.css}}
 
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{{Team:Lethbridge/assets/sidemenuremove.css}}
 
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<html>
 
<html>
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<head>
 
 
<style>
 
<style>
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/*SIDE MENU FOR IGEM STUFF*/
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/*RESET AND REMOVAL CSS IS PART OF RESET AND SIDEMENUREMOVE TEMPLATE*/
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#sideMenu{width: 200px;position: absolute;top: 20px;left: 1020px;z-index: 10;padding-top: 0px;padding-bottom: 15px;padding-left: 15px;padding-right: 15px;background-color: white;text-align: left;display: none;z-index: 9996;}
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/* RULES FOR ALL PAGES ARE PART OF THE ALLPAGES.CSS TEMPLATE*/
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#content{width: 100%;padding: 0px;margin-left: 0px;}
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/* RULES FOR THIS PAGE ONLY */
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#top_menu_14{height: 20px;}
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/*#globalWrapper{padding-bottom: 200px;}*/
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/*VISIBLE CONTENT STYLING BEGINS HERE*/
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body {padding-top: 3.9em;}
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</style>
 
</style>
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</head>
 
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<body>
 
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</body>
 
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</html>
 
  
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                <div class="content_box">
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<body>
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<div><img src="https://static.igem.org/mediawiki/2017/9/95/Banner_PAimproved.png"  class="bannerImg"></div>
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<div id="allContent">
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<br>
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<center>
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<form>
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    <input class="tealButton" type="button" value="MetRS" onclick="window.location.href='#anchor1'" />
 +
</form>
 
      
 
      
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                    <h1>Composite Parts</h1>
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<form>
 +
    <input class="tealButton" type="button" value="T7 RNAP" onclick="window.location.href='#anchor2'" />
 +
</form>
 
      
 
      
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                    <h2>Part BBa_K1791001</h2>
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<form>
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<p>Part BBa_K1791001 contains two modules, a standard ribosome binding site (RBS) expressing MS2 coat protein and a theophylline ribozyme with a MS2 aptamer at the 3’ end. To express MS2 coat  protein the part utilizes expression of c-terminal 6x His tagged MS2 coat protein under the regulation of  a T7 promoter and a standard Shine Dalgarno sequence followed by a double terminator (BBa_0015). Downstream of the RBS is a T7 promoter followed by a theophylline ribozyme and a MS2 aptamer at the  3’ end. Coexpression of the two modules results in MS2 binding to the 3’ end of the theophylline ribozyme module which allows for MS2-RNA purification by nickel affinity chromatography, RNA can  then be eluted from the MS2-RNA complex by addition of theophylline.  </p>
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    <input class="tealButton" type="button" value="EF-Ts" onclick="window.location.href='#anchor3'" />
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<img src="https://static.igem.org/mediawiki/2015/8/84/Uleth15_StandardRPA.png" height="30%" width="30%">
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</form>
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<p>Check it out on the <a href="http://parts.igem.org/Part:BBa_K1791001">Registry</a></p>
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 +
<form>
 +
    <input class="tealButton" type="button" value="EF-Tu" onclick="window.location.href='#anchor4'" />
 +
</form>
  
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                    <h2>Part BBa_K1791002</h2>
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<form>
−
<p>Part BBa_K17910012 contains two modules, a low efficiency ribosome binding site (RBS) expressing MS2 coat protein and a theophylline ribozyme with a MS2 aptamer at the 3’ end. To express  MS2 coat protein the part utilizes expression of c-terminal 6x His tagged MS2 coat protein under a T7 promoter and a standard Shine Dalgarno sequence followed by a double terminator (BBa_0015). Downstream of the RBS is a T7 promoter followed by a theophylline ribozyme and a MS2 aptamer at the 3’ end. Coexpression of the two modules results in MS2 binding to the 3’ end of the theophylline  ribozyme construct which allows for MS2-RNA purification by nickel affinity chromatography, RNA can  then be eluted from the MS2-RNA complex by addition of theophylline.</p>
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    <input class="tealButton" type="button" value="MS2BP" onclick="window.location.href='#anchor5'" />
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<img src="https://static.igem.org/mediawiki/2015/e/e5/Uleth_ConstructSchematic.png" height="30%" width="30%">
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</form>
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<p>Check it out on the <a href="http://parts.igem.org/Part:BBa_K1791002">Registry</a></p>
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<div style="clear:both" id="anchor1"></div>
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                </div>
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</center>
 +
 
 +
<br><br><br>
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<h2 align= "left"> Methionine Synthetase (MetRS)</h2>
 +
    <p class="text12j"> <b> Original part:</b> <a href="http://parts.igem.org/Part:BBa_K567015" id="pageLink" target="_blank">BBa_K567015</a>
 +
    <br><b>Submitted by:</b> SJTU-BioX-Shanghai 2011
 +
    <br><b>Designed by:</b> You Wang
 +
    <br><b>Rational behind improvements:</b>
 +
    BBa_K567015 encodes for a truncated MetRS without the anticodon recognition domain. We have improved this part by including the entire coding sequence, which has been codon optimized for optimal expression in <i>E. coli</i>. For easy purification we have included a C- terminal hexahistidine tag with a serine glycine linker. We have also optimized MetRS to be overexpressed in BL21 DE3 gold cell by putting it under the control of a T7 promoter (<a href="http://parts.igem.org/Part:BBa_I719005" id="pageLink" target="_blank">BBa_I719005</a>), RBS (<a href="http://parts.igem.org/Part:BBa_B0034" id="pageLink" target="_blank">BBa_B0034</a>) and double terminator (<a href="http://parts.igem.org/Part:BBa_B0015" id="pageLink" target="_blank">BBa_B0015</a>). The original part is also incompatible with biobrick standards. To address this we have removed all illegal cut sites from the construct.
 +
    <br><br>
 +
    <b>Our Part:</b>
 +
    </p>
 +
 
 +
<center>
 +
    <img src="https://static.igem.org/mediawiki/2017/4/4b/T--Lethbridge--MetRS.png" width=640px; height=138px; class="bannerImg" />
 +
</center>
 +
 
 +
<br><br>
 +
<div style="clear:both" id="anchor2"></div><br><br>
 +
<hr class="dividerLine">
 +
<br>
 +
 
 +
<h2 align= "left"> T7 RNA Polymerase (T7 RNAP)</h2>
 +
    <p class="text12j"> <b> Original part:</b> <a href="http://parts.igem.org/Part:BBa_I2032" id="pageLink" target="_blank">BBa_I2032</a>
 +
    <br><b>Submitted by:</b> MIT
 +
    <br><b>Designed by:</b> Bartholomew Canton
 +
    <br><b>Rational behind improvements:</b> BBa_I2032 encodes exclusively for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our N<i>ex</i>t <i>vivo</i> system we have Codon optimized for use in <i>E. coli</i> and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized our T7 RNA polymerase to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (<a href="http://parts.igem.org/Part:BBa_I719005" id="pageLink" target="_blank">BBa_I719005</a>), RBS (<a href="http://parts.igem.org/Part:BBa_B0034" id="pageLink" target="_blank">BBa_B0034</a>) and double terminator (<a href="http://parts.igem.org/Part:BBa_B0015" id="pageLink" target="_blank">BBa_B0015</a>).
 +
    <br><br>
 +
    <b>Our Part:</b>
 +
    </p>
 +
 
 +
<center>
 +
    <img src="https://static.igem.org/mediawiki/2017/0/07/T--Lethbridge--T7cs.png" width=640px; height=138px; class="bannerImg" />
 +
</center>
 +
 
 +
<br><br>
 +
<div style="clear:both" id="anchor3"></div><br><br>
 +
<hr class="dividerLine">
 +
<br>
 +
 
 +
<h2 align= "left"> Elongation Factor Thermo Stable (EF-Ts)</h2>
 +
    <p class="text12j"> <b> Original part:</b> <a href="http://parts.igem.org/Part:BBa_K1906000" id="pageLink" target="_blank">BBa_K1906000</a>
 +
    <br><b>Submitted by:</b> XJTLU-China 2016
 +
    <br><b>Designed by:</b> Wenbo Xu
 +
    <br><b>Rational behind improvements:</b> BBa_K1906000 encodes exclusively for the coding region of EF-Ts. In order to improve it and incorporate it into our N<i>ex</i>t <i>vivo</i> system we have codon optimized for use in <i>E. coli</i> and attached a C-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Ts to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (<a href="http://parts.igem.org/Part:BBa_I719005" id="pageLink" target="_blank">BBa_I719005</a>), RBS (<a href="http://parts.igem.org/Part:BBa_B0034" id="pageLink" target="_blank">BBa_B0034</a>) and double terminator (<a href="http://parts.igem.org/Part:BBa_B0015" id="pageLink" target="_blank">BBa_B0015</a>).
 +
    <br><br>
 +
    <b> Our Part:</b>
 +
    </p>
 +
 
 +
<center>
 +
    <img src="https://static.igem.org/mediawiki/2017/b/b1/T--Lethbridge--Ef-Ts.png" width=640px; height=138px; class="bannerImg" />
 +
</center>
 +
 
 +
   
 +
<br><br>
 +
<div style="clear:both" id="anchor4"></div><br><br>
 +
<hr class="dividerLine">
 +
<br>
 +
 
 +
<h2 align= "left"> Elongation Factor Thermo Unstable (EF-Tu)</h2>
 +
    <p class="text12j"><b> Original part:</b><a href="http://parts.igem.org/Part:BBa_K1906004" id="pageLink" target="_blank">BBa_K1906004</a>
 +
    <br><b>Submitted by:</b> XJTLU-China 2016
 +
    <br><b>Designed by:</b> Yuwei Han
 +
    <br><b> Rational behind improvements:</b> BBa_K1906004 encodes exclusively for the coding region of EF-Tu. In order to improve it and incorporate it into our N<i>ex</i>t <i>vivo</i> system we have codon optimized for use in <i>E. coli</i> and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Tu to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (<a href="http://parts.igem.org/Part:BBa_I719005" id="pageLink" target="_blank">BBa_I719005</a>), RBS (<a href="http://parts.igem.org/Part:BBa_B0034" id="pageLink" target="_blank">BBa_B0034</a>) and double terminator (<a href="http://parts.igem.org/Part:BBa_B0015" id="pageLink" target="_blank">BBa_B0015</a>).
 +
    <br><br>
 +
    <b> Our Part:</b>
 +
    </p>
 +
 
 +
<center>
 +
    <img src="https://static.igem.org/mediawiki/2017/f/f7/T--Lethbridge--Ef-Tu.png" width=640px; height=138px; class="bannerImg" />
 +
</center>
 +
 
 +
<br><br>
 +
<div style="clear: both" id="anchor5"></div><br><br>
 +
<hr class="dividerLine">
 +
<br>
 +
 
 +
<h2 align= "left">MS2 Coat Protein</h2>
 +
    <p class="text12j"><b> Original part:</b> <a href="http://parts.igem.org/Part:BBa_K2109108" id="pageLink" target="_blank">BBa_K2109108</a>
 +
    <br><b>Submitted by:</b> Lethbridge 2016
 +
    <br><b>Designed by:</b> Andy Hudson
 +
    <br><b>Rational behind improvements:</b> This year we utilized the MS2 coat protein (MS2BP) in our purification strategy of tRNA and ribosomes. As such, we expressed and purified MS2BP along with our tRNA<sup>Phe</sup> construct. From this we concluded that MS2BP expresses, binds to Nickel Sepharose affinity column, and binds the MS2 hairpin as designed.
 +
        <br><br>
 +
        <b> Our Part:</b>
 +
    </p>
 +
 
 +
<center>
 +
    <img src="https://static.igem.org/mediawiki/2017/b/b0/T--Lethbridge--MS2BP.png" width=640px; height=138px; class="bannerImg" />
 +
</center>
 +
 
 +
<br><br>
 +
<div style="clear: both"></div>
 +
 
 +
</div><!-- closing #allContent -->
 +
<img src="https://static.igem.org/mediawiki/2017/7/7d/Banner_footer_blank.png" class="bannerImg">
 +
</body>
 +
</html>

Latest revision as of 03:02, 2 November 2017

top




Methionine Synthetase (MetRS)

Original part: BBa_K567015
Submitted by: SJTU-BioX-Shanghai 2011
Designed by: You Wang
Rational behind improvements: BBa_K567015 encodes for a truncated MetRS without the anticodon recognition domain. We have improved this part by including the entire coding sequence, which has been codon optimized for optimal expression in E. coli. For easy purification we have included a C- terminal hexahistidine tag with a serine glycine linker. We have also optimized MetRS to be overexpressed in BL21 DE3 gold cell by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015). The original part is also incompatible with biobrick standards. To address this we have removed all illegal cut sites from the construct.

Our Part:






T7 RNA Polymerase (T7 RNAP)

Original part: BBa_I2032
Submitted by: MIT
Designed by: Bartholomew Canton
Rational behind improvements: BBa_I2032 encodes exclusively for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our Next vivo system we have Codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized our T7 RNA polymerase to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).

Our Part:






Elongation Factor Thermo Stable (EF-Ts)

Original part: BBa_K1906000
Submitted by: XJTLU-China 2016
Designed by: Wenbo Xu
Rational behind improvements: BBa_K1906000 encodes exclusively for the coding region of EF-Ts. In order to improve it and incorporate it into our Next vivo system we have codon optimized for use in E. coli and attached a C-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Ts to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).

Our Part:






Elongation Factor Thermo Unstable (EF-Tu)

Original part:BBa_K1906004
Submitted by: XJTLU-China 2016
Designed by: Yuwei Han
Rational behind improvements: BBa_K1906004 encodes exclusively for the coding region of EF-Tu. In order to improve it and incorporate it into our Next vivo system we have codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Tu to be overexpressed in BL21 DE3 gold cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).

Our Part:






MS2 Coat Protein

Original part: BBa_K2109108
Submitted by: Lethbridge 2016
Designed by: Andy Hudson
Rational behind improvements: This year we utilized the MS2 coat protein (MS2BP) in our purification strategy of tRNA and ribosomes. As such, we expressed and purified MS2BP along with our tRNAPhe construct. From this we concluded that MS2BP expresses, binds to Nickel Sepharose affinity column, and binds the MS2 hairpin as designed.

Our Part: