Photoregulation of in vitro transcription/translation of GFP by tethering an azobenzene to T7 promoter.

Abstract

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.