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HEK293细胞系:一种用于表达重组蛋白的载体。

HEK293 cell line: a vehicle for the expression of recombinant proteins.

作者信息

Thomas Philip, Smart Trevor G

机构信息

Department of Pharmacology, University College London, Gower Street, London, WC1E 6BT, UK.

出版信息

J Pharmacol Toxicol Methods. 2005 May-Jun;51(3):187-200. doi: 10.1016/j.vascn.2004.08.014.

Abstract

The HEK cell line has been extensively used as an expression tool for recombinant proteins since it was generated over 25 years ago. Although of epithelial origin, its biochemical machinery is capable of carrying out most of the post-translational folding and processing required to generate functional, mature protein from a wide spectrum of both mammalian and non-mammalian nucleic acids. Though popular as a transient expression system, this cell type has also seen wide use in stably transfected forms (i.e. transformed cells) to study a variety of cell-biological questions in neurobiology. The principal attributes which have made the HEK cell a popular choice among electrophysiologists to study isolated receptor channels include; its quick and easy reproduction and maintenance; amenability to transfection using a wide variety of methods; high efficiency of transfection and protein production; faithful translation and processing of proteins; and small cell size with minimal processes appropriate for voltage-clamp experimentation. These, and other attributes, also mean that complementary biochemical/cell biological evaluations of expressed proteins can be performed in concert with functional analyses to establish detailed pharmacological and biophysical profiles for the action of new drugs and their targets. The increased amount of sequence information available from the human genome has placed greater emphasis upon heterologous cell expression systems as targets for high throughput structure-function evaluation of novel drug targets and disease markers. Here we have highlighted some of the innate characteristics of the HEK cell in order that its suitability as a vehicle for the expression of a gene product can be assessed for particular needs. We have also detailed some of the standard methods used for transfection and obtaining functional data from electrophysiological recording techniques.

摘要

自25多年前产生以来,HEK细胞系已被广泛用作重组蛋白的表达工具。尽管它起源于上皮细胞,但其生化机制能够对来自多种哺乳动物和非哺乳动物核酸的功能性成熟蛋白进行大部分翻译后折叠和加工。作为一种瞬时表达系统,这种细胞类型在稳定转染形式(即转化细胞)中也有广泛应用,用于研究神经生物学中的各种细胞生物学问题。使HEK细胞成为电生理学家研究分离受体通道的热门选择的主要特性包括:其快速且易于繁殖和维持;易于使用多种方法进行转染;转染和蛋白生产效率高;蛋白的忠实翻译和加工;以及细胞体积小且突起最少,适合电压钳实验。这些特性以及其他特性还意味着,可以在进行功能分析的同时对表达的蛋白进行补充性生化/细胞生物学评估,以建立新药及其靶点作用的详细药理学和生物物理图谱。人类基因组中可用序列信息的增加,使得异源细胞表达系统作为新型药物靶点和疾病标志物高通量结构-功能评估的目标受到了更多关注。在此,我们强调了HEK细胞的一些固有特性,以便能够根据特定需求评估其作为基因产物表达载体的适用性。我们还详细介绍了一些用于转染以及从电生理记录技术中获取功能数据的标准方法。

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