SAP18和Sufu蛋白质复合物的表达纯化研究

Expression and Purifi cation of Suppressor of Fused in Complex with Sin3-Associated Polypeptide of 18 kDa

通过分子克隆技术将Sufu和SAP18构建到原核载体,在原核表达系统进行外源表达,采用亲和层析和分子筛层析对Sufu和SAP18及其复合物进行纯化。同时利用非变性胶的方法进一步确定该蛋白之间的相互作用及结合比例。结果表明,原核表达系统中Sufu和SAP18蛋白表达量高,可以纯化到高纯度的蛋白质。Sufu和SAP18结合的摩尔比为1:1,按摩尔比1:2混合、纯化可以得到高纯度、稳定的蛋白复合物,从而为进一步复合物的结构生物学研究奠定基础。

Molecular cloning, prokaryotic expression and protein purification technology were used to obtain human SAP 18, human Sufu and human SAP 18-Sufu protein complex. The native PAGE method was used to identify the interaction and the ratio between SAP18 and Sufu proteins. The result demonstrated that SAP 18 and Sufu proteins were highly expressed in the prokaryotic expression system. SAP 18 interacted with Sufu by a molar ratio of 1：1. The high purity and stability of the SAP 18-Sufu complex were obtained after the co-purification. These results laid a solid foundation for the future structural biology research on the protein complex.