title: Semi automated production of a set of different recombinant GST-Streptag fusion proteins.
abstract: We describe a high-throughput procedure for the large-scale production of recombinant GST-Streptag fusion proteins. This three-step process, comprising cloning, expression and purification, simultaneously produces up to 96 different proteins in a multi-well format with high yield and purity. Two complementary oligonucleotides, together encoding a specific peptide sequence are annealed and directly ligated into a pre-digested pGEX-2T plasmid carrying an N-terminal GST-tag and a C-terminal Streptag. Following expression, a multichannel pipetting robot purifies the resulting fusion proteins within 2 h by affinity chromatography on Streptactin Macroprep mini-columns.
authors.affiliation: Department of Protein Biochemistry, GPC-Biotech AG, Fraunhoferstr. 20, D-82152, Martinsried, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Sebastian
authors.forename: Jelena
authors.initials: J
authors.lastname: Wallwitz
authors.forename: Stefan
authors.initials: S
authors.lastname: Schmidt
mesh_terms: D001331:Automation; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017931:DNA Primers; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase