Team:NTNU Trondheim/Characterization

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Characterization Results

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 [[Openwetware prefix and suffix]http://openwetware.org/wiki/Synthetic_Biology:BioBricks/Part_fabrication], which contain E, X and S, P restriction sites.

To test the downregulating-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.