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Optogenetic control of Bacillus subtilis gene expression

Author

Listed:
  • Sebastian M. Castillo-Hair

    (Rice University)

  • Elliot A. Baerman

    (Rice University)

  • Masaya Fujita

    (University of Houston)

  • Oleg A. Igoshin

    (Rice University
    Rice University
    Rice University)

  • Jeffrey J. Tabor

    (Rice University
    Rice University)

Abstract

The Gram-positive bacterium Bacillus subtilis exhibits complex spatial and temporal gene expression signals. Although optogenetic tools are ideal for studying such processes, none has been engineered for this organism. Here, we port a cyanobacterial light sensor pathway comprising the green/red photoreversible two-component system CcaSR, two metabolic enzymes for production of the chromophore phycocyanobilin (PCB), and an output promoter to control transcription of a gene of interest into B. subtilis. Following an initial non-functional design, we optimize expression of pathway genes, enhance PCB production via a translational fusion of the biosynthetic enzymes, engineer a strong chimeric output promoter, and increase dynamic range with a miniaturized photosensor kinase. Our final design exhibits over 70-fold activation and rapid response dynamics, making it well-suited to studying a wide range of gene regulatory processes. In addition, the synthetic biology methods we develop to port this pathway should make B. subtilis easier to engineer in the future.

Suggested Citation

  • Sebastian M. Castillo-Hair & Elliot A. Baerman & Masaya Fujita & Oleg A. Igoshin & Jeffrey J. Tabor, 2019. "Optogenetic control of Bacillus subtilis gene expression," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-10906-6
    DOI: 10.1038/s41467-019-10906-6
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    Cited by:

    1. Ryan McQuillen & Amilcar J. Perez & Xinxing Yang & Christopher H. Bohrer & Erika L. Smith & Sylvia Chareyre & Ho-Ching Tiffany Tsui & Kevin E. Bruce & Yin Mon Hla & Joshua W. McCausland & Malcolm E. W, 2024. "Light-dependent modulation of protein localization and function in living bacteria cells," Nature Communications, Nature, vol. 15(1), pages 1-15, December.

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