greenTEV represents the catalytic domain of the nuclear inclusion a (NIa) protein with a molecular weight of 27 kDa encoded by the plant virus Tobacco Etch Virus. "green" indicates fusion of the protease to green fluorescent protein (GFP), which leads to increased stability and solubility of TEV protease. Moreover, greenTEV has been optimized by site directed mutagenesis to prevent autocatalytic cleavage. greenTEV is a highly site-specific cysteine protease that recognizes the amino acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) [ENLYFQ(G/S)] and cleaves between the residues Gln and Gly/Ser. The most commonly used recognition sequence is ENLYFQG. In biotechnology, greenTEV is a versatile enzyme to remove affinity tags from recombinant proteins with high specificity and activity over a wide range of pH, ionic strength and temperatures between 4 °C and 30 °C. The optimal temperature for cleavage is 30 °C. It is recommended to improve cleavage efficiency for each fusion protein by varying the amount of recombinant greenTEV, reaction time, or incubation temperature. The great advantage of greenTEV is its facile removal after cleavage reaction by immobilized metal affinity chromatography (IMAC) since it is equipped with a His-tag. Furthermore, the removal of greenTEV can be monitored instantly by detection of fluorescence in solution - this easy, fast and sensitive method omits time-consuming SDS-PAGE or Western blot analysis. If the green fluorescence of greenTEV is not suitable and you prefer blue fluorescence as readout, check our blueTEV protease.
