Effective photoregulation of gene expression by photo responsive T7 promoter

We constructed a photoresponsive T7 promoter by tethering two 2,6-dimethyl azobenzenes (4-(2,6-dimethyl-phenlylazo)-benzoic acids) via D-threoninol linkers. Under UV light irradiation, the rate of transcription with T7 RNA polymerase (RNAP) on the photoresponsive promoter was 10-fold faster than tha...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic Acids Symposium Series 2007-11, Vol.51 (1), p.349-350
Main Authors: Liang, Xingguo, Wakuda, Ryuji, Tsuda, Yuichiro, Nishioka, Hidenori, Asanuma, Hiroyuki
Format: Article
Language:English
Online Access:Request full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We constructed a photoresponsive T7 promoter by tethering two 2,6-dimethyl azobenzenes (4-(2,6-dimethyl-phenlylazo)-benzoic acids) via D-threoninol linkers. Under UV light irradiation, the rate of transcription with T7 RNA polymerase (RNAP) on the photoresponsive promoter was 10-fold faster than that under visible light irradiation. Kinetic analysis revealed that Km of cis-cis-promoter (both of the introduced azobenzenes are in cis-form) with T7 RNAP was more than 60 times larger than that of trans-trans-form, indicating that the high photoregulatory efficiency was obtained mainly due to the remarkable difference in their affinity with RNAP. By attaching a photoresponsive promoter to the GFP gene, we also showed that photoregulation of gene expression became practicable.
ISSN:0261-3166
1746-8272
DOI:10.1093/nass/nrm175