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.