Heterologous Expression of Membrane Proteins in E. coli.
Heterologous Expression of Membrane Proteins in E. coli.
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膜蛋白在大肠杆菌中的异源表达。
DOI：
10.1007/978-1-0716-2368-8_4
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Depping P
中科院分区：
文献类型：
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作者：
Depping P
几十年来，大肠杆菌（Escherichia coli，E. coli）已经成为重组蛋白生产的基石，用于多种膜蛋白的异源合成。由于其在廉价培养基中生长迅速，密度高，易于操作和处理，E。大肠杆菌是一系列膜蛋白靶的优良宿主细胞。此外，其遗传易处理性允许筛选各种基因构建体以获得最佳表达条件，从而在短时间内产生相对高的膜蛋白产量。在这里，我们描述了一般的工作流程生产膜蛋白在E。杆菌我们提供的方案显示了感兴趣的基因如何被修饰，转移到表达载体和宿主中，以及如何优化和分析膜蛋白产量。我们举例说明的例子非常适合那些开始进入膜蛋白生产世界的科学家。
Over the decades, the bacteriumEscherichia coli(E. coli)has become the cornerstone of recombinant protein production, used for heterologous synthesis of a variety of membrane proteins. Due to its rapid growth to high densities in cheap media, and its ease of manipulation and handling,E. coliis an excellent host cell for a range of membrane protein targets. Furthermore, its genetic tractability allows for a variety of gene constructs to be screened for optimal expression conditions, resulting in relatively high yields of membrane protein in a short amount of time. Here, we describe the general workflow for the production of membrane proteins inE. coli. The protocols we provide show how the gene of interest is modified, transferred to an expression vector and host, and how membrane protein yields can be optimized and analyzed. The examples we illustrate are well suited for scientists who are starting their journey into the world of membrane protein production.