Synthetic microbial consortia enable rapid assembly of pure translation machinery

Fernando Villarreal, Luis E. Contreras-Llano, Michael Chavez, Yunfeng Ding, Jinzhen Fan, Tingrui Pan, Cheemeng Tan

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Assembly of recombinant multiprotein systems requires multiple culturing and purification steps that scale linearly with the number of constituent proteins. This problem is particularly pronounced in the preparation of the 34 proteins involved in transcription and translation systems, which are fundamental biochemistry tools for reconstitution of cellular pathways ex vivo. Here, we engineer synthetic microbial consortia consisting of between 15 and 34 Escherichia coli strains to assemble the 34 proteins in a single culturing, lysis, and purification procedure. The expression of these proteins is controlled by synthetic genetic modules to produce the proteins at the correct ratios. We show that the pure multiprotein system is functional and reproducible, and has low protein contaminants. We also demonstrate its application in the screening of synthetic promoters and protease inhibitors. Our work establishes a novel strategy for producing pure translation machinery, which may be extended to the production of other multiprotein systems.

Original languageEnglish (US)
Pages (from-to)29-35
Number of pages7
JournalNature Chemical Biology
Volume14
Issue number1
DOIs
StatePublished - Jan 1 2018

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Villarreal, F., Contreras-Llano, L. E., Chavez, M., Ding, Y., Fan, J., Pan, T., & Tan, C. (2018). Synthetic microbial consortia enable rapid assembly of pure translation machinery. Nature Chemical Biology, 14(1), 29-35. https://doi.org/10.1038/nchembio.2514