[Construction and characterization of TetR and GFP fusion protein].
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 (TR::GFP) was purified. The binding of TR::GFP with tetracycline (tc) was demonstrated by nitrocellulose filter binding assay. TR::GFP also maintained the fluorescence property of GFP. Most significantly, fluorescence emission intensity of TR::GFP 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 TR::GFP possesses characteristics of a tetracycline biosensor.
Zuo
Yan
Y
State Key Laboratoy of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Yang
Ke-Qian
KQ
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D037401:Scyphozoa / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013752:Tetracycline / Q000378:metabolism*; D013753:Tetracycline Resistance
D001426:Bacterial Proteins; D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C083139:Tet O resistance protein, Bacteria; C034811:tetracycline resistance-encoding transposon repressor protein; D049452:Green Fluorescent Proteins; D013752:Tetracycline