Effective photoregulation of gene expression by photo responsive T7 promoter | Zendy

Xingguo Liang | Zendy; R. Wakuda | Zendy; Y. Tsuda | Zendy; H. Nishioka | Zendy; Hiroyuki Asanuma | Zendy

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Effective photoregulation of gene expression by photo responsive T7 promoter

Author(s) -

Xingguo Liang,

R. Wakuda,

Y. Tsuda,

H. Nishioka,

Hiroyuki Asanuma

Publication year - 2007

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/nrm175

Subject(s) - t7 rna polymerase , transcription (linguistics) , chemistry , gene , rna polymerase , irradiation , promoter , gene expression , rna polymerase ii , biophysics , microbiology and biotechnology , stereochemistry , rna , biochemistry , biology , linguistics , philosophy , physics , escherichia coli , bacteriophage , nuclear physics

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 K(m) 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.

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