酵母中线粒体膜蛋白复合物的克隆与多亚基表达。
Cloning and Multi-Subunit Expression of Mitochondrial Membrane Protein Complexes in Saccharomyces cerevisiae.
机构信息
Laboratory of Molecular Biology, National Institute of Diabetes & Digestive & Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
Vertex Pharmaceuticals, Boston, MA, USA.
出版信息
Methods Mol Biol. 2020;2127:1-11. doi: 10.1007/978-1-0716-0373-4_1.
Saccharomyces cerevisiae is a useful eukaryotic expression system for mitochondrial membrane proteins due to its ease of growth and ability to provide a native membrane environment. The development of the pBEVY vector system has further increased the potential of S. cerevisiae as an expression system by creating a method for expressing multiple proteins simultaneously. This vector system is amenable to the expression and purification of multi-subunit protein complexes. Here we describe the cloning, yeast transformation, and co-expression of multi-subunit outer mitochondrial membrane complexes using the pBEVY vector system.
酿酒酵母是一种有用的真核表达系统，可用于线粒体膜蛋白，因为其易于生长并且能够提供天然的膜环境。pBEVY 载体系统的开发通过创建一种同时表达多种蛋白质的方法，进一步增加了酿酒酵母作为表达系统的潜力。该载体系统适用于多亚基蛋白复合物的表达和纯化。本文描述了使用 pBEVY 载体系统克隆、酵母转化和多亚基外线粒体膜复合物的共表达。
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