Construction and characterization of TetR and GFP fusion protein / 生物工程学报

ABSTRACT

Tetracycline repressor gene (tetR) from E. coli transposon Tn10 was fused in frame with green fluorescent protein gene (gfp) from jellyfish Aequorea Victoria on an E. coli expression vector and the fusion protein (TRGFP) was purified. The binding of TRGFP with tetracycline (tc) was demonstrated by nitrocellulose filter binding assay. TRGFP also maintained the fluorescence property of GFP. Most significantly, fluorescence emission intensity of TRGFP increased by 2-fold in the presence of tc, from 1.132 to 2.214, while those of GFP and TetR showed little change under similar conditions. The results indicated TRGFP possesses characteristics of a tetracycline biosensor.