細胞および分子生物学

抽象的な

Impressive outcome of recombinant protein purification applied to GSTtag affinity chromatography: A review

Dr. Asaduaman1 *, Lutfun Nahar2 , Md. Bakhtiar Lijon3 , Shehen Imrn4 and Mohammad Saedur Rhamane5

Affinity Tags have been performed as the potential tools in the basic biological research field especially for production of recombinant protein and functional proteomics. Those affinity tags were wildly applied to simplify the purification of recombinant protein as well as differentiation of protein complex. Glutathione-S-Transferase (GST) tag has been extensively used in affinity chromatography for purification of fusion/recombinant protein to analysis of structure and function of protein, protein-protein interaction and to produce pharmaceutical product. In this review we describe the advantage of GST-tag in affinity chromatography technique as a method for inducible, high level protein expression and purification of recombinant protein. Recombinant protein which is expressed in a pGEX or pET vectors and that protein with GST-tag encoded at the NH2 - or COOH- region of genome sequence. There are some expression vectors which has different site to approve for unidirectional insertion of the coding region DNA, promoter, primers, antibiotic, Ori and GST-tag into pGEX vectors. By used reduced glutathione (GSH) during affinity chromatography the GST-tag with recombinant protein is eluted and stored it. Displacement of the GST-tag from the recombinant protein performed by protease enzyme for digestion which is purified by the application of another affinity technique.