Cold Spring Harb Protoc. 2021 Jan 4;2021(1):2021/1/pdb.prot102186. doi: 10.1101/pdb.prot102186.
Recovery of intracellular proteins requires disruption of the host cell before the target protein is extracted and isolated. Disruption methods vary depending on the type of cells, the total volume, and the number of samples being processed. For cells enveloped in cell walls (such as yeast), mild techniques such as hypotonic shock are not sufficient to achieve adequate lysis. More vigorous methods are often required. Although the preferred medium- or large-scale method of breaking yeast cells is mechanical shearing, lysis with the aid of glass beads in a BeadBeater is described here.
恢复细胞内蛋白质需要在提取和分离目标蛋白质之前破坏宿主细胞。破坏方法取决于细胞类型、总体积和正在处理的样本数量。对于被细胞壁包围的细胞(如酵母),低渗休克等温和技术不足以实现充分的裂解。通常需要更剧烈的方法。虽然机械剪切是破碎酵母细胞的首选中规模或大规模方法,但此处介绍了在珠式研碎仪中使用玻璃珠辅助裂解的方法。