title: Construction of a compatible Gateway-based co-expression vector set for expressing multiprotein complexes in E. coli.
abstract: We report the construction of a versatile Gateway-based co-expression vector set for producing multiprotein complexes in Escherichia coli. The set consists of two groups of three vectors (pCoGW and pCo0GW), each having a specific antibiotic resistance gene, a compatible origin of replication and allowing cloning of up to two genes, each under control of its own T7 promoter. To validate the set, 33 (co-)expression plasmids encoding fluorescent protein (GFP, DsRed and ECFP) have been generated. Protein expression levels were quantified and (co-)expression visualized by fluorescent microscopy. The results illustrate the applicability of these vectors in co-expression studies.
authors: Service de Biologie Moléculaire, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/Inserm/Université de Strasbourg, 1 rue Laurent Fries, BP10142, 67404 Illkirch cedex, France.
authors: Loubna
authors: L
authors: Salim
authors: Service de Biologie Moléculaire, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/Inserm/Université de Strasbourg, 1 rue Laurent Fries, BP10142, 67404 Illkirch cedex, France.
authors: Claire
authors: C
authors: Feger
authors: Service de Biologie Moléculaire, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/Inserm/Université de Strasbourg, 1 rue Laurent Fries, BP10142, 67404 Illkirch cedex, France. Electronic address: didier.busso@cea.fr.
authors: Didier
authors: D
authors: Busso
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D046912:Multiprotein Complexes / Q000096:biosynthesis / Q000235:genetics
chemical_list: D046912:Multiprotein Complexes; D049452:Green Fluorescent Proteins
keywords: Co-expression; Expression screening; Gateway; Protein complex; Recombinational cloning; Versatility