重组MBP-SUV39H1在大肠杆菌中表达与纯化

Expression and purification of MBP-SUV39H1 protein in E. coli

[目的]在大肠杆菌中获得具有甲基转移酶活性的重组MBP-SUV39H1蛋白。[方法]通过在大肠杆菌中同时表达异染色质蛋白1(HP1)与重组MBP-SUV39H1蛋白的方法,实现了MBP-SUV39H1的表达,采用his亲和纯化与分子筛Superdex200(SD200)两步分离纯化方案,并利用质谱和ELISA检测MBP-SUV39H1的甲基转移酶活性。[结果]利用大肠杆菌成功表达了MBP-SUV39H1融合蛋白,经纯化后目的条带单一,并具有良好的甲基转移酶活性。[结论]纯化的具有甲基转移酶活性的MBP-SUV39H1可用于抑制剂筛选等后续研究。

[Objective]To get recombinant MBP-SUV39 H1 protein with methyltransferase activity in BL21.[Methods]The expression of fusion protein MBP-SUV39 H1 was completed by coexpressing it with Heterochromatin Protein 1( HP1) in BL21,His-affinity resin and molecular sieve Superdex200( SD200) two-step purification protocol was established,the methyltransferase activity of MBP-SUV39 H1 was assayed by MS and Elisa.[Result]The recombinant MBP-SUV39 H1 protein was successfully expressed in E. coli,the purified MBP-SUV39 H1 had a high purity with single band,and a good methyltransferase activity.[Conclusion]The purified MBP-SUV39 H1 could be used for further study.