title: Insect cells-baculovirus system for the production of difficult to express proteins.
abstract: The production of sufficient quantities of homogenous protein not only is an essential prelude for structural investigations but also represents a rate-limiting step for many human functional studies. Although technologies for expression of recombinant proteins and complexes have been improved tremendously, in many cases, protein production remains a challenge and can be associated with considerable investment. This chapter describes simple and efficient protocols for expression screening and optimization of protein production in insect cells using the baculovirus expression system. We describe the procedure, starting from the cloning of a gene of interest into an expression transfer baculovirus vector, followed by generation of the recombinant virus by homologous recombination, evaluation of protein expression, and scale-up. Handling of insect cell cultures and preparation of bacmid for co-transfection are also detailed.
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS, BP 163, Illkirch, Cedex 67404, France.
authors.forename: Judit
authors.initials: J
authors.lastname: Osz-Papai
authors.forename: Laura
authors.initials: L
authors.lastname: Radu
authors.forename: Wassim
authors.initials: W
authors.lastname: Abdulrahman
authors.forename: Isabelle
authors.initials: I
authors.lastname: Kolb-Cheynel
authors.forename: Nathalie
authors.initials: N
authors.lastname: Troffer-Charlier
authors.forename: Catherine
authors.initials: C
authors.lastname: Birck
authors.forename: Arnaud
authors.initials: A
authors.lastname: Poterszman
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007313:Insecta / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods
chemical_list: D011994:Recombinant Proteins