@article {Nguyen027656, author = {Phuc H.B. Nguyen and Yong Y. Wu and Shaobin Guo and Richard M. Murray}, title = {Design Space Exploration of the Violacein Pathway in Escherichia coli Based Transcription Translation Cell-Free System (TX-TL)}, elocation-id = {027656}, year = {2015}, doi = {10.1101/027656}, publisher = {Cold Spring Harbor Laboratory}, 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{\textquoteright}s coding sequence (CDS) significantly affected its expression level. Finally, pathway exploration suggested an improvement in violacein production at high VioC and VioD DNA concentrations.}, URL = {https://www.biorxiv.org/content/early/2015/09/27/027656}, eprint = {https://www.biorxiv.org/content/early/2015/09/27/027656.full.pdf}, journal = {bioRxiv} }