Difference between revisions of "Part:BBa K396000:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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===Applications of BBa_K396000===
 
===Applications of BBa_K396000===
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<br><br><br>
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<font size="6" T7/CI-OR1 hybrid promoter</font>&nbsp;&nbsp;&nbsp;<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K396000">BBa_K396000</a><br>
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&nbsp;&nbsp;&nbsp; low unregulated, repressed by lambda CI, activated by T7 RNA Polymerase, repression is stronger than activation.<br><br>
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<table border="0" cellpadding="30" cellspacing="0">
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  <td width="800px"><font size="5" face=verdana>T7/CI-OR1 hybrid promoter</font><br>
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  <hr width="500" size="1" align="left"><br>
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  <font size="2" face=verdana>
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T7/CI-OR1 hybrid promoter was designed to be activated by T7 RNA Polymerase and repressed by lambda CI protein. In sequence design,  lambda CI operator site 1 (OR1, Part of BBa_R1051) was directly attached to the downstream of T7 promoter sequence (BBa_I719005). This characterization was implemented to validate the promoter function. In results, the activation and repression was actually observed.
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<br>
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</font></td> 
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</table>
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<table border="0" cellpadding="30" cellspacing="0">
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<tr>
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  <td width="800px"><font size="5" face=verdana>Experiments</font><br>
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  <hr width="500" size="1" align="left"><br>
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</tr>
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</table>
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<img src="https://static.igem.org/mediawiki/2010/1/10/T7-CI-1.png">
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<table border="0" cellpadding="30" cellspacing="0">
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<tr><td>
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<br><br>
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  <td width="800px"><font size="5" face=verdana>Results and Conclusion</font><br>
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<font size="2" face=verdana>
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Under the situation of CI non-expressed , T7 RNAP causes GFP expression by activating T7/CI hybrid
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promoter (Fig.1). On the other hand, under the situation of CI expressed, T7 RNAP does not cause GFP
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expression (Fig.2). The reason of this is considered as following.  Under the situation of CI expressed,
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even though T7 RNAP tries to  activate T7/CI promoter, the promoter Is repressed by CI because the
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ability of CI repression is superior to the one of T7 activation for this promoter.  So it is concluded that
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T7/CI hybrid  promoter is to be activated by T7 RNA Polymerase and repressed by lambda  CI protein,
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following the rules of CI repression > T7 activation.   
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</font></td> 
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</tr>
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</table>
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<img src="https://static.igem.org/mediawiki/2010/b/bd/CI-2.png">
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<img src="https://static.igem.org/mediawiki/2010/7/73/CI-3.jpg">
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<table border="0" cellpadding="30" cellspacing="0">
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  <td width="800px"><font size="5" face=verdana>Reference</font><br>
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<font size="3">
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<br>1)
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<br>2)
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</td>
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</td>
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===User Reviews===
 
===User Reviews===

Revision as of 14:14, 27 October 2010

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K396000




   BBa_K396000
    low unregulated, repressed by lambda CI, activated by T7 RNA Polymerase, repression is stronger than activation.

T7/CI-OR1 hybrid promoter

T7/CI-OR1 hybrid promoter was designed to be activated by T7 RNA Polymerase and repressed by lambda CI protein. In sequence design, lambda CI operator site 1 (OR1, Part of BBa_R1051) was directly attached to the downstream of T7 promoter sequence (BBa_I719005). This characterization was implemented to validate the promoter function. In results, the activation and repression was actually observed.
Experiments



Results and Conclusion

Under the situation of CI non-expressed , T7 RNAP causes GFP expression by activating T7/CI hybrid promoter (Fig.1). On the other hand, under the situation of CI expressed, T7 RNAP does not cause GFP expression (Fig.2). The reason of this is considered as following. Under the situation of CI expressed, even though T7 RNAP tries to activate T7/CI promoter, the promoter Is repressed by CI because the ability of CI repression is superior to the one of T7 activation for this promoter. So it is concluded that T7/CI hybrid promoter is to be activated by T7 RNA Polymerase and repressed by lambda CI protein, following the rules of CI repression > T7 activation.
Reference


1)
2)

User Reviews

UNIQ9bc6059b627446c5-partinfo-00000001-QINU UNIQ9bc6059b627446c5-partinfo-00000002-QINU