重组蛋白包涵体的复性研究

Renaturation of the Inclusion Bodies Within Recombinant Protein

重组蛋白在大肠杆菌中的高表达往往形成不可溶、无生物活性的包涵体,需经过变性溶解后,在适当条件下复性形成天然的构象,才可恢复其生物活性.变复性实验是建立在对蛋白质体外折叠机制的了解的基础上.根据近年来对蛋白质折叠机制的认识和重组蛋白包涵体在复性方面的主要进展,论述以下3个方面的内容1)蛋白质在细胞内的折叠机制;2)蛋白质体外折叠机制;3)蛋白质复性的策略和方法.

Overexpression of recombinant proteins in the Escherichia coli cytoplasm often results in the formation of insoluble inclusion bodies.Active protein can be recovered by solubilization of inclusion bodies followed by renaturation of the solubilized protein.The process of renaturation is established in the understanding to the mechanism of protein folding in vitro.According to the progression of protein folding research,three questions are reviewed:1) the protein folding in vivo;2)the mechanism of protein folding in vitro;3) the renaturation strategy.