System-level studies of a cell-free transcription-translation platform for metabolic engineering

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Title System-level studies of a cell-free transcription-translation platform for metabolic engineering
Authors Yong Y. Wu, Hirokazu Sato, Hongjun Huang, Stephanie J. Culler, Julia Khandurina, Harish Nagarajan, Tae Hoon Yang, Stephen Van Dien, Richard M. Murray
Source bioRxiv Technical Report
Abstract Current methods for assembling biosynthetic pathways in microorganisms require a process of repeated trial and error and have long design-build-test cycles. We describe the use of a cell-free transcription-translation (TX-TL) system as a biomolecular breadboard for the rapid engineering of the 1,4-butanediol (BDO) pathway. We demonstrate the reliability of TX-TL as a platform for engineering biological systems by undertaking a careful characterization of its transcription and translation capabilities and provide a detailed analysis of its metabolic output. Using TX-TL to survey the design space of the BDO pathway enables rapid tuning of pathway enzyme expression levels for improved product yield. Leveraging TX-TL to screen enzyme variants for improved catalytic activity accelerates design iterations that can be directly applied to in vivo strain development.
Type Technical report
URL http://www.biorxiv.org/content/early/2017/08/03/172007
Tag wu+17-biorxiv
ID 2017i
Funding GBMF PMTI
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