Construction of a compatible Gateway-based co-expression vector set for expressing multiprotein complexes in E. coli

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• 作者：Loubna Salim¹ ; Claire Feger² ; Didier Busso ; ^{didier.busso@cea.fr" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Co-expression ; Expression screening ; Gateway ; Protein complex ; Recombinational cloning ; Versatility
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：1 November 2016
• 年：2016
• 卷：512
• 期：Complete
• 页码：110-113
• 全文大小：1003 K

文摘

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.