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Team:BostonU/Multiplexing
From 2014.igem.org
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As the key method of Phase II of the Chimera workflow, multiplexing is being employed by our group to determine the full range of function of our transcriptional units (TUs). We define full range of function as having a set of TUs with one varied constituent part, whose transfer curves display a difference across at least two decades of REU with varying peak fluorescence as measured by Stanton et al..[1] This wide range is desirable and essential to the robustness of the workflow because it provides the constitutive parts of a device with diverse expression levels. Having this diversity makes having a desired combination of parts more likely, which can more easily result in a desired device behavior. We have selected multiplexing as our tool to achieve this diversity because of its compatibility with our MoClo assembly method, which facilitates the quick replacement of any set of parts in a transcriptional unit. Our first multiplexing reactions vary the ribosomal binding sites (RBSs) of transcriptional units. |
References[1] B. Stanton, A. Nielsen, A. Tamsir, K. Clancy, T. Peterson & C. Voigt (2014). "Genomic mining of prokaryotic repressors for orthogonal logic gates." Nature Chemical Biology 10: 99-105. doi:10.1038/nchembio.1411 |
