Team:NTNU Trondheim/Characterization

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Characterization Results

Several experiments were performed to test our BioBricks and constructs. This page will try to give a short summary of the construction of the test-constructs, the experiments performed to test them, and the results from the experiments.

ppGpp's effect on rrnB P1 promoter

To be able to use the rrnB P1 promoter, most of the BioBrick BBa_K112118 (lacking the first 11 bp) was amplified with PCR. The primers were designed to give the [http://openwetware.org/wiki/Synthetic_Biology:BioBricks/Part_fabrication Openwetware prefix and suffix]Openwetware prefix and suffix, which contain E, X and S, P restriction sites.

To test the down-regulating effect of ppGpp on rrnB P1, which is the basis of our project, we decided to make a construct containing ppGpp Synthase (RelA) inducible by the pBAD/AraC promoter. The construct would also contain beta-galactosidase (lacZ) which would be expressed by the rrnB P1 promoter. Thus, induction of the pBAD/AraC promoter with arabinose should give lower lacZ production, as relA is overproduced giving high ppGpp concentration.