Photoregulation of in vitro transcription/translation of GFP by tethering an azobenzene to T7 promoter.
Based on a previous study of photo-regulation of transcription by T7 RNA polymerase, the regulatory effect of azobenzene on in vitro green fluorescent protein (GFP) production was studied by introduction of a D-threoninol-tethered p-azobenzene into the non template strand of the T7 promoter in a GFP template. By measurement of the resulting GFP fluorescence intensity after the in vitro transcription/translation coupled reaction, it is found that photo-regulation of GFP gene expression can be achieved in a position-dependent manner by trans-cis isomerization of azobenzene upon UV irradiation.
Zhao
Jing
J
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1, Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Zhou
Jingmin
J
Liu
Mingzhe
M
Asanuma
Hiroyuki
H
Komiyama
Makoto
M
D001391:Azo Compounds / Q000378:metabolism*; D017123:Bacteriophage T7 / Q000235:genetics; D001483:Base Sequence; D005786:Gene Expression Regulation / Q000528:radiation effects; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics*; D008027:Light; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014176:Protein Biosynthesis / Q000528:radiation effects*; D013698:Templates, Genetic; D014158:Transcription, Genetic / Q000528:radiation effects*
D001391:Azo Compounds; D049452:Green Fluorescent Proteins; C009850:azobenzene