Testing usu.html

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Revision as of 06:25, 17 October 2010

Utah State University

CyanoBricks

Testing

Promoter Expression Testing

Promoter activation was tested by constructing a protein expression device using the BioBrick promoter, the Synechocystis RBS (BBa_M11402), the GFP(mut3b) (BBa_E0040), and terminator sequences (BBa_B0015) in the vector (BBa_M11400) in E. coli. Once the device was constructed, plasmid DNA was extracted and Synechocystis was naturally transformed, using the protocol described in Zang, et al. (2007). Baseline expression readings were taken by diluting a sample of cells to the same O.D. using a spectrophotometer, and reading the GFP fluorescence with a spectrofluorometer under normal, non-stressing conditions. Stress was then induced, and fluorescence was re-measured. All fluorescence readings were converted to relative fluorescence compared to GFP expression driven by the sigA promoter under the same treatment conditions.

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            <td><a class="sideLinks" href="Construction_usu.html" title="Construction"><img src="https://static.igem.org/mediawiki/2010/3/37/Construction.png" height="60"></a></td>
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-           <td><a class="sideLinks" href="Modeling_usu.html" title="Software Modeling"><img src="https://static.igem.org/mediawiki/2010/3/3c/Modeling.png" height="60"></a></td>
+           <!td><!a class="sideLinks" href="Modeling_usu.html" title="Software Modeling"><!img src="https://static.igem.org/mediawiki/2010/3/3c/Modeling.png" height="60"></a></td>
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            <td><a class="sideLinks" href="Testing_usu.html" title="Testing"><img src="https://static.igem.org/mediawiki/2010/7/7f/Testing.png" height="60"></a></td>
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-Add some information for testing....
+Promoter Expression Testing
+<hr>
+Promoter activation was tested by constructing a protein expression device using the BioBrick promoter, the Synechocystis RBS (BBa_M11402), the GFP(mut3b) (BBa_E0040), and terminator sequences (BBa_B0015) in the vector (BBa_M11400) in E. coli. Once the device was constructed, plasmid DNA was extracted and Synechocystis was naturally transformed, using the protocol described in Zang, et al. (2007). Baseline expression readings were taken by diluting a sample of cells to the same O.D. using a spectrophotometer, and reading the GFP fluorescence with a spectrofluorometer under normal, non-stressing conditions. Stress was then induced, and fluorescence was re-measured. All fluorescence readings were converted to relative fluorescence compared to GFP expression driven by the sigA promoter under the same treatment conditions.