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Team:NTNU Trondheim/Results
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| - | ==Characterization of ppGpp effect on the rrnB P1 promoter== | + | ==Characterization of ppGpp-effect on the rrnB P1 promoter== |
To be able to use the rrnB P1 promoter, most of it (lacking the first 11 bp) was amplified with PCR from the BioBrick BBa_K112118. The primers were designed to give the openwetware prefix and suffix, which contain E, X and S, P restriction sites. | To be able to use the rrnB P1 promoter, most of it (lacking the first 11 bp) was amplified with PCR from the BioBrick BBa_K112118. The primers were designed to give the openwetware prefix and suffix, which contain E, X and S, P restriction sites. | ||
Latest revision as of 09:47, 19 August 2011
Results
Characterization of ppGpp-effect on the rrnB P1 promoter
To be able to use the rrnB P1 promoter, most of it (lacking the first 11 bp) was amplified with PCR from the BioBrick BBa_K112118. The primers were designed to give the openwetware prefix and suffix, 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.
