Testing usu.html

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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.

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.