title: Transient transfection coupled to baculovirus infection for rapid protein expression screening in insect cells.
abstract: Baculovirus infected insect cells are widely used for heterologous protein expression. Despite the power of this system, the use of baculovirus techniques for protein expression screening is hampered by the time and resources needed to generate each recombinant baculovirus. Here, we show that a transfection/infection based expression system is suitable for screening of expression constructs in insect cells and represents a valid alternative to other traditional screening methodologies using recombinant baculovirus. The described method is based on gene delivery by transfection coupled to the induction of protein expression by non-recombinant baculovirus infection. Vectors that control expression by a combination of the baculovirus promoters ie1 and p10 and the enhancer element hr5 are among the ones suitable for this method. Infection with non-recombinant baculovirus drastically increases the basal activity of these elements, leading to protein over-expression. Multiple vectors can be simultaneously co-transfected/infected, making transfection/infection amenable for screening of multiple co-expressed proteins and protein complexes. Taken together, our results prove that the transfection/infection protocol is a valid and innovative approach for increasing speed and reducing costs of protein expression screening for structural and functional studies.
authors.affiliation: Dept. of Biochemistry and Molecular Biology, Center for Brain Research, Medical University of Vienna, Spitalgasse 4 1090 Vienna, Austria.
authors.forename: Stephan
authors.initials: S
authors.lastname: Radner
authors.forename: Patrick H N
authors.initials: PH
authors.lastname: Celie
authors.forename: Karoline
authors.initials: K
authors.lastname: Fuchs
authors.forename: Werner
authors.initials: W
authors.lastname: Sieghart
authors.forename: Titia K
authors.initials: TK
authors.lastname: Sixma
authors.forename: Mariano
authors.initials: M
authors.lastname: Stornaiuolo
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D014162:Transfection / Q000379:methods*
chemical_list: D011994:Recombinant Proteins