Preparation of insoluble GST fusion proteins.
A wide variety of plasmid vectors are commercially available for the production of fusion proteins in bacterial cells. Most are also designed to incorporate a tag that allows affinity purification of the expressed fusion protein from bacterial cell extracts. The most commonly used are vectors that incorporate a portion of the glutathione-S-transferase (GST) enzyme that is able to bind to immobilized glutathione and vectors that use a polyhistidine tag which binds immobilized nickel ions with high affinity. This protocol describes preparation of an insoluble GST fusion protein, isolated using glutathione agarose beads.
Ilaria
I
Rebay
Richard G
RG
Fehon
D001671:Biochemistry / Q000379:methods*; D002458:Cell Fractionation; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D012995:Solubility
D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase