Team:NRP-UEA-Norwich/Quanticare
From 2012.igem.org
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- | The comparator circuit project involves multiple promoters that are sensitive to similar chemicals, a different reporter/effector enzyme for each promoter, and a range of specially designed DNA leaders that go between the promoter and reporter of each gene system. The idea involves the specially designed DNA leaders being complementary to one another and resulting in DNA annealing which covers the ribosome binding site; this means that mRNAs transcribed following activation of the promoters will cancel one another out and fail to be translated. If there is an imbalance of substrates one set of mRNAs would be produced in excess of the others | + | The comparator circuit project involves multiple promoters that are sensitive to similar chemicals, a different reporter/effector enzyme for each promoter, and a range of specially designed DNA leaders that go between the promoter and reporter of each gene system. The idea involves the specially designed DNA leaders being complementary to one another and resulting in DNA annealing which covers the ribosome binding site; this means that mRNAs transcribed following activation of the promoters will cancel one another out and fail to be translated. If there is an imbalance of substrates one set of mRNAs would be produced in excess of the others as this set would not anneal to one another, therefore resulting in translation and expression of the associated protein. This is demonstrated in ''Figure 1.'' where two sets of mRNA have been produced, one with an RFP reporter and one with a CFP reporter; due to an imbalance in substrates four lots of the RFP-associated mRNA have been produced, while only two lots of the CFP-associated mRNAs have been produced. As the image shows the two CFP-associated mRNAs have annealed with two of the RFP-associated mRNAs, blocking the RBS and preventing translation, however due to the imbalance there are two RFP-associated mRNAs left to be translated and thus RFP would be expressed in this situation. |
Revision as of 14:34, 26 September 2012