Expression and Purification of SH2 Domains Using Baculovirus Expression System

Methods Mol Biol. 2017:1555:183-198. doi: 10.1007/978-1-4939-6762-9_11.

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.

Keywords: Bac-to-Bac; Baculovirus expression system; GST fusion protein; Insect cells; SH2 domain.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cloning, Molecular
  • Gene Expression*
  • Gene Order
  • Genetic Vectors / genetics*
  • Humans
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / isolation & purification*
  • Sf9 Cells
  • Transfection
  • Workflow
  • src Homology Domains*

Substances

  • Recombinant Fusion Proteins