【摘要】 目的建立一套高纯度、高活性表达糖原合成酶激酶3β的技术。方法分别采用大肠杆菌和杆状病毒昆虫细胞蛋白表达系统表达糖原合成酶激酶3β,通过His和GST标签两步纯化目的蛋白,SDS-PAGE观察目的蛋白的纯度,激酶反应观察目的蛋白的活性。结果大肠杆菌表达纯化的糖原合成酶激酶3β占纯化后总蛋白的量为54%,昆虫细胞表达纯化的糖原合成酶激酶3β占纯化后总蛋白的量为96%;昆虫细胞来源的目的蛋白比大肠杆菌来源的目的蛋白活性高一倍左右。结论相比大肠杆菌表达的糖原合成酶激酶3β,昆虫细胞表达的目的蛋白具有更高的纯度和活性。更多还原

【Abstract】 Objective To establish a method for the expression of glycogen synthase kinase 3β with high purity and biological activity.Methods E.coli expression system and baculovirus-insect cell expression system were used to produce the kinase,followed by purification using His-tag and GST-tag and determination of its purity and activity by SDS-PAGE and kinase reaction,respectively.Results Glycogen synthase kinase 3β produced from E.coli represented 54% of the total bacterial protein,as compared with 96% of the total protein from the insect cell system.Glycogen synthase kinase 3β produced from insect cell exhibited an one-fold higher biological activity than the protein obtained from E.coli.Conclusions Compared with the protein from E.coli system,glycogen synthase kinase 3β from the insect cell expression system is endowed with a higher purity and bioactivity.更多还原