In the sphere of fundamental biological research, affinity
tags have been used as potentially useful instruments, particularly for the
synthesis of recombinant proteins and functional proteomics. These affinity
tags were widely used to make the differentiation of protein complexes and the
purification of recombinant proteins easier. Affinity chromatography has made
substantial use of glutathione-S-Transferase (GST) tag for fusion/recombinant
protein purification, protein-protein interaction research, and the production
of medicinal products. 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. A recombinant protein is a
protein which is expressed in 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 sites to approve for unidirectional
insertion of the coding region DNA, promoter, primers, antibiotic, Ori and
GST-tag into pGEX vectors. Reduced glutathione (GSH) is utilized in affinity
chromatography to elute and store the GST-tag containing recombinant protein.
The recombinant protein's GST tag is removed for digestion using a protease
enzyme, and the protein is then purified using a different affinity approach.
Author(s)details:-
Md
Asaduzzaman
Department of Bacteriology, Graduate School of Medicine, Dentistry and
Pharmaceutical Sciences, Okayama University, Okayama, 700-8558, Japan.
Lutfun
Nahar
Department of General Medicine, Graduate School of Medicine, Dentistry
and Pharmaceutical Sciences, Okayama University Hospital, Okayama University,
700-8558, Japan.
Please See
the book here :- https://doi.org/10.9734/bpi/ibs/v5/12804F
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