Simpson Richard J
Cold Spring Harb Protoc. 2010 Sep 1;2010(9):pdb.prot5485. doi: 10.1101/pdb.prot5485.
The high levels of expression produced by molecular cloning techniques make bacteria particularly useful for producing recombinant proteins. However, these proteins often are difficult to purify, owing to their tendency to aggregate and precipitate within the bacteria as insoluble inclusion bodies. Formation of inclusion bodies is especially common for nonbacterial proteins. Although no single method can be applied to every protein, a number of strategies are available to solubilize inclusion body proteins. This protocol describes one such method.
分子克隆技术所产生的高水平表达使得细菌在生产重组蛋白方面特别有用。然而,这些蛋白质往往难以纯化,因为它们倾向于在细菌内聚集并沉淀为不溶性包涵体。包涵体的形成在非细菌蛋白中尤为常见。虽然没有一种方法能适用于每种蛋白质,但有许多策略可用于溶解包涵体蛋白。本方案描述了其中一种方法。