首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Construction of a compatible Gateway-based co-expression vector set for expressing multiprotein complexes in E-coli
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Construction of a compatible Gateway-based co-expression vector set for expressing multiprotein complexes in E-coli

机译:用于在大肠杆菌中表达多蛋白复合物的兼容的基于Gateway的共表达载体集的构建

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摘要

We report the construction of a versatile Gateway-based co-expression vector set for producing multi protein complexes in Escherichia coli. The set consists of two groups of three vectors (pCoGW and pCoOGW), 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. (C) 2016 Elsevier Inc. All rights reserved.
机译:我们报告了一套通用的基于网关的共表达载体的构建,用于在大肠杆菌中生产多种蛋白质复合物。该组由两组组成,每组三个载体(pCoGW和pCoOGW),每个载体具有特定的抗生素抗性基因,兼容的复制起点,并允许克隆最多两个基因,每个基因都在其自己的T7启动子的控制下。为了验证该组,已产生了33个(共)表达质粒,它们编码荧光蛋白(GFP,DsRed和ECFP)。定量蛋白质表达水平,并通过荧光显微镜观察(共)表达。结果说明了这些载体在共表达研究中的适用性。 (C)2016 Elsevier Inc.保留所有权利。

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