Team:Tokyo Tech/Construction

From 2008.igem.org

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== '''Construction''' ==
== '''Construction''' ==
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[[Image:Construction.png|260px|thumb|left|figure1 We constructed A.PtetR-GFP, B.promoter less-GFP for confirmatory experiment]]
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For confirming pressure-response ability of pressure-inducible promoter, we experimented  
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under 30MPa pressure. We chose TetR promoter(PtetR) as a pressure-inducible promoter.  
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we constructed two plasmids - one is PtetR-GFP on pSB6([http://partsregistry.org/Part:BBa_K121010 BBa_K121010]), the other is promoter less-GFP on pSB6([http://partsregistry.org/Part:BBa_K121013 BBa_K121013]).
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<p class=MsoNormal>For checking pressure-response ability of Plac, we constructed three plasmids.Those are showed at figure 1.Plac stands for a promoter repressed by lacI protein. Ptet stands for promoter repressed by tetR protein.∆p stands for promoter less.JM109 is constitutive expression lacI protein so that Plac is always repressed by lacI protein.And JM109 isn’t expression tetR. Therefore, we used Ptet as a positive control.  </font></p>
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<p class=MsoNormal>Fig1</p></td>
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<IMG src="https://static.igem.org/mediawiki/2008/c/c8/Tech_const_fig1.jpg
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<p class=MsoNormal>Each parts is constructed on low-copy plasmid <a herf=http://partsregistry.org/Part:BBa_I739201>“pSB6”</a> ,and inserted into E.coli strain JM109.</p></td>
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For confirming pressure-response ability of pressure-inducible promoter, we  
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experimented under 30MPa pressure. We chose TetR promoter(Ptet) as a pressure-inducible promoter. we constructed two plasmids - one is Ptet-GFP on pSB6([http://partsregistry.org/Part:BBa_K121010 BBa_K121010]), the other is promoter less-GFP on pSB6([http://partsregistry.org/Part:BBa_K121013 BBa_K121013]).
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[[Image:Ptet-GFP.png|250px|center|fig1.Ptet-GFP]]
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[[Image:promoter_less-GFP.png|180px|center|fig1.Promoter less-GFP]]
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Revision as of 06:54, 26 October 2008

Home Construction Acrylic container Development of promoter Genetic toggle switch Parts Submitted to the Registry Result The Team Notebook

Construction

figure1 We constructed A.PtetR-GFP, B.promoter less-GFP for confirmatory experiment

For confirming pressure-response ability of pressure-inducible promoter, we experimented under 30MPa pressure. We chose TetR promoter(PtetR) as a pressure-inducible promoter. we constructed two plasmids - one is PtetR-GFP on pSB6([http://partsregistry.org/Part:BBa_K121010 BBa_K121010]), the other is promoter less-GFP on pSB6([http://partsregistry.org/Part:BBa_K121013 BBa_K121013]).

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