title: Expression and Purification of SH2 Domains Using Baculovirus Expression System.
abstract: Recombinant proteins expressed in bacteria are sometimes insoluble, aggregated, and incorrectly folded. For those Src homology 2 (SH2) domains that are insoluble in bacteria, baculovirus-insect cell expression systems can be an alternative to produce soluble and functionally active proteins. We describe a protocol for cloning and purification of GST-tagged SH2 domains using the Bac-to-Bac baculovirus expression system.
authors.affiliation: Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, 400 Farmington Avenue, Farmington, CT, 06032, USA.
authors.forename: Mari
authors.initials: M
authors.lastname: Ogiue-Ikeda
authors.affiliation: Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, 400 Farmington Avenue, Farmington, CT, 06032, USA. machida@uchc.edu.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Machida
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D061987:Sf9 Cells; D014162:Transfection; D057188:Workflow; D018909:src Homology Domains
chemical_list: D011993:Recombinant Fusion Proteins
keywords: Bac-to-Bac; Baculovirus expression system; GST fusion protein; Insect cells; SH2 domain