• Medientyp: E-Artikel
  • Titel: Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG
  • Beteiligte: Binder, Dennis [VerfasserIn]; Grünberger, Alexander [VerfasserIn]; Loeschcke, Anita [VerfasserIn]; Probst, Christopher [VerfasserIn]; Bier, Claus [VerfasserIn]; Pietruszka, Jörg [VerfasserIn]; Wiechert, Wolfgang [VerfasserIn]; Kohlheyer, Dietrich [VerfasserIn]; Jaeger, Karl-Erich [VerfasserIn]; Drepper, Thomas [VerfasserIn]
  • Erschienen: RSC Publ., 2014
  • Erschienen in: Integrative biology 6(8), 755-765 (2014). doi:10.1039/c4ib00027g
  • Sprache: Englisch
  • DOI: https://doi.org/10.1039/c4ib00027g
  • ISSN: 1757-9694; 1757-9708
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  • Beschreibung: Light can be used to control numerous cellular processes including protein function and interaction as well as gene expression in a non-invasive fashion and with unprecedented spatiotemporal resolution. However, for chemical phototriggers tight, gradual, and homogeneous light response has never been attained in living cells. Here, we report on a light-responsive bacterial T7 RNA polymerase expression system based on a photocaged derivative of the inducer molecule isopropyl-β-D-thiogalactopyranoside (IPTG). We have comparatively analyzed different Escherichia coli lac promoter-regulated expression systems in batch and microfluidic single-cell cultivation. The lacY-deficient E. coli strain Tuner(DE3) harboring additional plasmid-born copies of the lacI gene exhibited a sensitive and defined response to increasing IPTG concentrations. Photocaged IPTG served as a synthetic photo-switch to convert the E. coli system into an optogenetic expression module allowing for precise and gradual light-triggering of gene expression as demonstrated at the single cell level.
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