Journal Article

FZJ-2014-03427

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Light-responsive control of bacterial gene Expression: precise triggering of the lac promoter activity using photocaged IPTG

Binder, D.FZJ* ; Grünberger, A.FZJ* ; Loeschcke, A.FZJ* ; Probst, C.FZJ* ; Bier, C.FZJ* ; Pietruszka, J.FZJ* ; Wiechert, W.FZJ* ; Kohlheyer, D.FZJ* ; Jaeger, K.-E. (Corresponding Author)FZJ* ; Drepper, T.FZJ*

2014
RSC Publ. Cambridge

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Please use a persistent id in citations: http://hdl.handle.net/2128/7981  doi:10.1039/c4ib00027g

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.

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Contributing Institute(s):

1. Biotechnologie (IBG-1)
2. Institut für Bioorganische Chemie (HHUD) (IBOC)

Research Program(s):

1. 899 - ohne Topic (POF2-899) (POF2-899)

Appears in the scientific report 2014

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Database coverage:
; ; ; BIOSIS Previews ; JCR ; NCBI Molecular Biology Database ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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