Edit Paper: Design Space Exploration of the Violacein Pathway in Escherichia coli Based Cell-Free System

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Title*:	Design Space Exploration of the Violacein Pathway in Escherichia coli Based Cell-Free System
Authors*:	Phuc H.B. Nguyen, Yong Wu, Shaobin Guo, Richard M Murray
Source*:	Submitted, 2016 Synthetic Biology: Engineering, Evolution and Design (SEED) Conference
Abstract*:	In this study, an Escherichia coli (E. coli) based transcription translation cell-free system (TX-TL) was employed to sample various enzyme expression levels of the violacein pathway. TX-TL enables rapid modifications and prototyping of the pathway without complicated cloning cycles. The violacein metabolic pathway has been successfully reconstructed in TX-TL. Analysis of the product via UV-Vis absorption and liquid chromatography-mass spectrometry detected 4.95 mM of violacein. Expression levels of pathway enzymes were modeled using the TX-TL Toolbox. The model revealed the length of an enzyme coding sequence (CDS) significantly affected its expression level. Finally, pathway exploration suggested an improvement in violacein production at high VioC and VioD DNA concentrations.
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