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Issue 8, 2014
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Light-responsive control of bacterial gene expression: precise triggering of the lac promoter activity using photocaged IPTG

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Abstract

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.

Graphical abstract: Light-responsive control of bacterial gene expression: precise triggering of the lac promoter activity using photocaged IPTG

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Supplementary files

Article information


Submitted
07 Feb 2014
Accepted
15 May 2014
First published
16 May 2014

This article is Open Access

Integr. Biol., 2014,6, 755-765
Article type
Paper
Author version available

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