Multiprotein complex production in insect cells by using polyproteins

Methods Mol Biol. 2014:1091:131-41. doi: 10.1007/978-1-62703-691-7_8.

Abstract

A powerful approach utilizing polyproteins for balancing stoichiometry of recombinant multiprotein complexes overproduced in baculovirus expression vector systems (BEVS) is described. This procedure has been implemented here in the MultiBac system but can also be directly adapted to all commonly used BEVS. The protocol details the design principles of polyprotein-expressing DNA constructs, the generation of composite baculovirus for polyprotein production, and the expression and in vivo processing of polyproteins in baculovirus infected insect cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cell Line
  • Cloning, Molecular
  • Gene Order
  • Genetic Vectors / genetics
  • Homologous Recombination
  • Integrases / genetics
  • Integrases / metabolism
  • Multiprotein Complexes / biosynthesis*
  • Multiprotein Complexes / genetics*
  • Plasmids / genetics
  • Polyproteins / biosynthesis*
  • Polyproteins / genetics*
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics*

Substances

  • Multiprotein Complexes
  • Polyproteins
  • Recombinant Proteins
  • Cre recombinase
  • Integrases