Part:BBa_J100097:Experience

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Applications of BBa_J100097

We inserted this promoter, tetO, into J100091 upstream of RFP. This plasmid is ampicillin resistant. We transformed E. coli to contain our plasmid with tetO and then grew the cells on LB+amp plates. Then, we grew overnight cultures of a small colony divided into one culture of LB+amp+anhydrotetracycline and one of LB+amp. We grew another overnight culture of a big colony divided into LB+amp+anhydrotetracycline and LB+amp. For our positive control, we grew E. coli containing a known promoter, PlacI , and prepared an overnight culture in LB+amp. For our negative control we grew E. coli that contained our plasmid and the transcription terminator and prepared an overnight culture in LB+amp.

We measured the fluorescence for each population of cells in our overnight cultures and divided it by absorbance of each sample to correct for variances in population density. We found that our promoter worked better without anhydrotetracycline and worked almost as well as the known promoter PlacI.

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