Katzke Nadine, Bergmann René, Jaeger Karl-Erich, Drepper Thomas
Institute of Molecular Enzyme Technology, Heinrich-Heine-University, Juelich, Germany.
Methods Mol Biol. 2012;824:251-69. doi: 10.1007/978-1-61779-433-9_13.
The functional expression of heterologous genes in standard hosts such as Escherichia coli is often hampered by various limitations including improper folding, incomplete targeting, and missassembly of the corresponding enzymes. This observation led to the development of numerous expression systems that are based on alternative, metabolic versatile hosts. One such organism is the Gram-negative phototrophic nonsulfur purple bacterium Rhodobacter capsulatus. During photosynthetic growth, R. capsulatus exhibits several unique properties including the formation of an intracytoplasmic membrane system as well as the synthesis of various metal-containing cofactors. These properties make R. capsulatus a promising expression host particularly suited for difficult-to-express proteins such as membrane proteins. In this chapter, we describe a novel R. capsulatus expression system and its application.
在诸如大肠杆菌等标准宿主中,异源基因的功能性表达常常受到各种限制的阻碍,这些限制包括相应酶的折叠不当、靶向不完整和组装错误。这一观察结果促使人们开发了许多基于替代性、代谢多功能宿主的表达系统。其中一种生物是革兰氏阴性光合非硫紫色细菌荚膜红细菌。在光合生长过程中,荚膜红细菌表现出几种独特的特性,包括形成胞内膜系统以及合成各种含金属的辅因子。这些特性使荚膜红细菌成为一种有前途的表达宿主,特别适合表达诸如膜蛋白等难以表达的蛋白质。在本章中,我们描述了一种新型的荚膜红细菌表达系统及其应用。