Construction of a compatible Gateway-based co-expression vector set for expressing multiprotein complexes in E. coli |
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Authors: | Loubna Salim Claire Feger Didier Busso |
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Institution: | 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 |
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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. |
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Keywords: | Co-expression Expression screening Gateway Protein complex Recombinational cloning Versatility |
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