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- Method for Assaying the Activity of Substrate Specific Proteases Utilizing Ecotin Fusion Protein
- Method for Assaying the Activity of Substrate Specific Proteases Utilizing Ecotin Fusion Protein
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机译:-利用Ecotin融合蛋白测定底物特异性蛋白酶活性的方法
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摘要
The present invention relates to a method measuring the activity of the specific enzyme for a specific substrate proteins. ; The present invention (1) fusing the echo tin carboxy terminus of a polypeptide comprising a portion that is cut by a specific enzyme for a specific substrate proteins and amino-terminal has GST, GFP, any one of the proteins selected from the group consisting of MBP phase by fusion to obtain a fusion protein; (2) the particular protease substrates and substrate to the fusion protein as a substrate, - performing an enzymatic reaction; Specific degradation of certain substrate proteins comprising the step of separating the supernatant by centrifugation and then heat the reaction of the enzyme reaction, and measuring the trypsin activity inhibition in the supernatant and (3) the substrate It provides a method of measuring the activity of the enzyme. ; When using the activity measuring method of the specific enzyme for a specific substrate proteins of the invention can be rapidly on fluorescence measuring via the suppression degree of the trypsin activity and to simply and measuring the effective activity of the specific enzyme for a specific substrate proteins. ; Ubiquitin, ubiquitin-like proteins, proteolytic enzymes, Eco tin, trypsin
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