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| | ===Applications of BBa_K396000=== | | ===Applications of BBa_K396000=== |
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| | <!--- Main Contents Start ---> | | <!--- Main Contents Start ---> |
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| − | <br><br><br>
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| − | <font size="6" T7/CI-OR1 hybrid promoter</font> <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K396000">BBa_K396000</a><br>
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| − | 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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| − | <tr>
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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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| − | </font></td>
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| − | </tr>
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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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| − | <hr width="500" size="1" align="left"><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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| − | <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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| − | <hr width="500" size="1" align="left"><br>
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| − | <font size="3">
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| − | <br>Dubendorf, J.W. & Studier, F.W. Controlling basal expression in an
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| − | inducible T7 expression system by blocking the target T7 promoter with
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| − | lac repressor. Journal of Molecular Biology 219, 45-59 (1991).
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| − | </td>
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| − | </td>
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| − | </table>
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| − | </html>
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| | ===User Reviews=== | | ===User Reviews=== |
UNIQ187f0b8a4b733e83-partinfo-00000000-QINU
UNIQ187f0b8a4b733e83-partinfo-00000001-QINU
