Team:TCU Taiwan/Parts

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       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Team"><font size="3">Members</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Team"><font size="3">Members</font></a></li>
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      <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Achievements"><font size="3">Achievements</font></a></li>
       <li><a href="https://igem.org/Team.cgi?id=1473" target="_blank"><font size="3">Official Team Profile</font></a></li>
       <li><a href="https://igem.org/Team.cgi?id=1473" target="_blank"><font size="3">Official Team Profile</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Contact"><font size="3">Contact</font></a></li>
       <li><a href="https://2014.igem.org/Team:TCU_Taiwan/Contact"><font size="3">Contact</font></a></li>
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   <td colspan="2" height="10px"><img src="https://static.igem.org/mediawiki/2014/b/b8/TCU-Phage.png" width="35px"/><font face="Trebuchet MS" size="5" color="#90B849">BBa_K1473005</font></td>
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   <td height="10px">&nbsp;</td>
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  <td valign="top" rowspan="4" align="center"><div style="width: 520px; padding: 0em">
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<img alt=" change width 586 to 520 & 745 " style="float: left; position: relative; z-index: 2" src="https://static.igem.org/mediawiki/2014/0/0f/TCU_PART_10718959_854093597942524_327155245_o.jpg" onmouseover="this.style.width='745px'" onmouseout="this.style.width='520px'" width="100%"><font size="3" face="Verdana"><strong>Fig.1</strong></font>
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   <td><img src="https://static.igem.org/mediawiki/2014/b/b8/TCU-Phage.png" alt="" width="35px"/><font face="Trebuchet MS" size="5" color="#90B849">BBa_K1473005</font></td>
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   <td valign="top" width="40%"><font size="3" face="Verdana" color="#333">Phagemid pBluescript is a  special plasmid, it functions as a normal plasmid when transformed into  bacteria. But when helper phage M13KO7 infect the bacteria, it will produce  progeny M13 phage as vector for pBluescript.&nbsp;<br><br>
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     pBluescript contains a f1 ori while M13KO7  helper phage&rsquo;s f1 ori has been knock-down. So after the infection of M13KO7,  its genome can produce protein and replicate but these proteins will package  pBluescript as their true &ldquo;genome&rdquo;. As a result, the progeny phage&rsquo;s genome  will only contains pBluescript&rsquo;s MCS, they cannot make next generation.</font></td>
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    <table width="90%" border="1" cellspacing="0" cellpadding="0">
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   <td valign="top" width="40%"><font size="3" face="Verdana" color="#333">Phagemid pBluescript is a  special plasmid, it functions as a normal plasmid when transformed into  bacteria. But when helper phage M13KO7 infect the bacteria, it will produce  phagemid-carrying phages.&nbsp;<br><br>
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        <td align="center"><table width="100%" border="1" cellspacing="0" cellpadding="0">
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     pBluescript contains a f1 ori while M13KO7  helper phage&rsquo;s f1 ori has been mutated. So after the infection of M13KO7,  its genome can produce protein and replicate but these proteins will package  pBluescript as &quot;genome&quot;. As a result, the pahemid-carrying phaes will only contain pBluescript, they cannot make next generation.</font></td>
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            <td align="center"><img src="https://static.igem.org/mediawiki/2014/0/0f/TCU_PART_10718959_854093597942524_327155245_o.jpg" width="90%"></td>
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            <td align="center"><font size="3" face="Verdana"><strong>Fig.1</strong></font></td>
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   <td valign="top"><font size="3" face="Verdana" color="#333">Here we combined a CRISPR system(BBa_K1218011) with an inducible promoter(BBa_K914003). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal  
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   <td valign="top"><font size="3" face="Verdana" color="#333">Here we combined a CRISPR system(<a href="http://parts.igem.org/Part:BBa_K1218011" target="_blank">BBa_K1218011</a>) with an inducible promoter(<a href="http://parts.igem.org/Part:BBa_K914003" target="_blank">BBa_K914003</a>). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal  
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     CRISPR array, it functions to knockout target gene. We give this system an inducible promoter so we can decided when to switch if on or off. </font></td>
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     CRISPR array, it functions to knockout target gene. We give this system an inducible promoter so we can decided when to switch it on or off. </font></td>
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         <td align="center"><img src="https://static.igem.org/mediawiki/2014/c/c7/TCU_CRISPR_POT.jpg" alt="" width="30%"></td>
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         <td align="center"><img src="https://static.igem.org/mediawiki/2014/c/c7/TCU_CRISPR_POT.jpg" alt="" width="60%"></td>
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           <td align="center"><img src="https://static.igem.org/mediawiki/2014/7/7c/TCU_PART_10736069_854704547881429_1390079281_n.jpg" width="100%"></td>
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           <td align="center"><img src="https://static.igem.org/mediawiki/2014/b/bf/TCU_SD_10733196_853652871319930_1466704235_o.jpg" width="100%"></td>
           <td align="center"><img src="https://static.igem.org/mediawiki/2014/b/bf/TCU_SD_10733196_853652871319930_1466704235_o.jpg" width="100%"></td>
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      <p>We put a <em>sac</em>I restriction  cutting site downstream to gRNA (not shown in sequence information) and ligated  it back to pSB1C3 with correct prefix and surffix. Then we cut this recombinant  plasmid with <em>sac</em>I and run gel  electrophoresis for testing. As we can see in this image, because pSB1C3 only  contains 1 <em>sac</em>I cutting site itself,  so it would not be cut into 2 parts when no gRNA is ligated inside. But if  there is, then <em>sac</em>I will cut this  recombinant plasmid into 2 parts, whose lengths are 939 bp and 1159 bp.  Obviously, we have successfully synthesized this gRNA.</p></font></td>
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      <p><font color="#333" size="3" face="Verdana">We put a <em>sac</em>I restriction  cutting site downstream to gRNA (not shown in sequence information) and ligated  it back to pSB1C3 with correct prefix and surffix. Then we cut this recombinant  plasmid with <em>sac</em>I and run gel  electrophoresis for checking. As we can see in this image, because pSB1C3 only  contains 1 <em>sac</em>I cutting site itself,  so it would not be cut into 2 parts when no gRNA is ligated inside. But if  there is, then <em>sac</em>I will cut this  recombinant plasmid into 2 parts, whose lengths are 939 bp and 1159 bp.  Obviously, we have successfully synthesized this gRNA.</font></p></td>
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Latest revision as of 21:09, 17 October 2014

 
Parts
  
Favorite parts
 
change width 586 to 520 & 745 Fig.1
BBa_K1473005  
   
Phagemid pBluescript is a special plasmid, it functions as a normal plasmid when transformed into bacteria. But when helper phage M13KO7 infect the bacteria, it will produce phagemid-carrying phages. 

pBluescript contains a f1 ori while M13KO7 helper phage’s f1 ori has been mutated. So after the infection of M13KO7, its genome can produce protein and replicate but these proteins will package pBluescript as "genome". As a result, the pahemid-carrying phaes will only contain pBluescript, they cannot make next generation.
BBa_K1473005
 
 
BBa_K1473009
 
Here we combined a CRISPR system(BBa_K1218011) with an inducible promoter(BBa_K914003). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal CRISPR array, it functions to knockout target gene. We give this system an inducible promoter so we can decided when to switch it on or off.
Source :Science

BBa_K1473009
 
 
 
Parts Sandbox
 
Click on the name of the parts for detailed information.
 
NameTypeDescriptionDesignerLength
  BBa_K1473001DNAgRNA 1 Shan, Zhe22
  BBa_K1473002DNAgRNA 2Shan, Zhe22
  BBa_K1473003DNAgRNA 3Shan, Zhe22
  BBa_K1473004DNAgRNA 4Shan, Zhe22
  BBa_K1473005Plasmid_Backbonephagemid pBluescript Shan, Zhe2929
  BBa_K1473006DNAgRNA for ampR 1Shan, Zhe22
  BBa_K1473007DNAgRNA for ampR 2Shan, Zhe22
  BBa_K1473008DNAgRNA for ampR 3Shan, Zhe22
 BBa_K1473009CodingpRha induced CRISPRShan, Zhe5011
  BBa_K1473010DNAgRNA for ampR 4Shan, Zhe22
  BBa_K1473011DNAgRNA for ampR 5Shan, Zhe22
  BBa_K1473012DNAgRNA for NeoR/kanR 1Shan, Zhe22
  BBa_K1473013DNAgRNA for NeoR/kanR 2Shan, Zhe22
  BBa_K1473014DNAgRNA for NeoR/kanR 3Shan, Zhe22
  BBa_K1473015DNAgRNA for NeoR/kanR 4Shan, Zhe22
  BBa_K1473016DNAgRNA for NeoR/kanR 5Shan, Zhe22
  BBa_K1473017DNAgRNA for tetR 1Shan, Zhe22
  BBa_K1473018DNAgRNA for tetR 2Shan, Zhe22
  BBa_K1473019DNAgRNA for tetR 3Shan, Zhe22
  BBa_K1473020DNAgRNA for tetR 4Shan, Zhe22
  BBa_K1473021DNAgRNA for tetR 5Shan, Zhe22
TCU_Taiwan 2014 iGEM Team Parts
gRNA
 
Fig.2
 
Fig.3

We put a sacI restriction cutting site downstream to gRNA (not shown in sequence information) and ligated it back to pSB1C3 with correct prefix and surffix. Then we cut this recombinant plasmid with sacI and run gel electrophoresis for checking. As we can see in this image, because pSB1C3 only contains 1 sacI cutting site itself, so it would not be cut into 2 parts when no gRNA is ligated inside. But if there is, then sacI will cut this recombinant plasmid into 2 parts, whose lengths are 939 bp and 1159 bp. Obviously, we have successfully synthesized this gRNA.
 
Parts Table

This table is from the Database of iGEM.
Any parts our team has created will appear in this table below:
<groupparts>iGEM014 TCU_Taiwan</groupparts>
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Team Members Project Parts Human Pratics Modeling Safety Notebook Attributions

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