title: Multiprotein complex production in insect cells by using polyproteins.
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.
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.
authors.forename: Yan
authors.initials: Y
authors.lastname: Nie
authors.forename: Itxaso
authors.initials: I
authors.lastname: Bellon-Echeverria
authors.forename: Simon
authors.initials: S
authors.lastname: Trowitzsch
authors.forename: Christoph
authors.initials: C
authors.lastname: Bieniossek
authors.forename: Imre
authors.initials: I
authors.lastname: Berger
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D059765:Homologous Recombination; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000096:biosynthesis* / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D020815:Polyproteins / Q000096:biosynthesis* / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D046912:Multiprotein Complexes; D020815:Polyproteins; D011994:Recombinant Proteins; C045073:Cre recombinase; D019426:Integrases