2型糖尿病是一种常见的慢性疾病,其发生与胰岛素的耐受相关。β-抑制蛋白2(β-arrestin2)的缺失或功能紊乱,会导致胰岛素抵抗和2型糖尿病的发生。而使用β-arrestin2进行治疗可明显缓解葡萄糖不耐受及胰岛素抵抗。本研究通过重组PCR方法,将小分子泛素相关修饰物(small ubiquitin-related modifier,SUMO)与β-arrestin2基因融合,再将融合基因插入原核表达载体p ET22b中,成功构建了p ET22b-SUMO-β-arrestin2表达载体,将重组表达载体转入大肠杆菌Rosetta Blue(DE3)宿主细胞,使用IPTG在温度为20℃诱导表达,再将诱导表达后的蛋白经Ni-NTA进行纯化,成功获得SUMO-β-arrestin2蛋白,为后续β-arrestin2药效学的研究奠定基础。
Type 2 diabetes is acommon chronic disease, and it is associated with insulin resistance. Loss or dysfunction of β-arrestin2 results in the development of insulin resistance and progression of type 2 diabetes. However, administration of β-arrestin2 significantly amend glucose intolerance and insulin resistance. In this article, We fused SUMO and β-arrestin2 gene with recombinant PCR methods, and then inserted it into pET22b vector, and constructed a expression vector pET22b-SUMO-β-arrestin2 successfully. A recombinant expression plasmid was expressed in Rosetta Blue (DE3) line of E.coli with IPTG at the temperature of 20 ℃. The SUMO-β-arrestin2 fusion protein was purified with Ni-NTA successfully. Lay the foundation for subsequent pharmacodynamic research of β-arrestin2.