谷氨酰胺tRNA合成酶的克隆、表达、纯化及活性测定  

Moleculor cloning,expression,purification and activity of glutaminyl-tRNA synthetase

作　　者：张银山[1] 韩宏岩[1] 许维岸[1] 

机构地区：[1]苏州大学基础医学与生物科学学院,苏州215123

出　　处：《生物学杂志》2016年第1期6-8,共3页Journal of Biology

基　　金：国家自然科学基金项目(No.81071306)

摘　　要：为了研究谷氨酰胺tRNA合成酶的结构和性质,人工合成谷氨酰胺tRNA合成酶基因,构建原核表达载体p PET-28a-His-GlnRS和p PIDTp SMART-tRNAGln表达载体,共转化到大肠杆菌BL21(DE3),经IPTG诱导表达,分离纯化谷氨酰胺tRNA合成酶,测定其活性。结果显示,共转化到大肠杆菌BL21(DE3)获得了可溶性的、有活性的目的蛋白,谷氨酰胺对谷氨酰胺tRNA合成酶没有底物抑制作用。In order to study the structure and function of glutaminyl-tRNA synthetase,the glutaminyl-tRNA synthetase and-tRNAGln gene was synthesized and subcloned into expression vector PET-28a-His-GlnRS and PIDTp SMART,separately,and then GlutaminyltRNA Synthetase protein in BL21（ DE3） was expressed by IPTG induction. The activity of purify Glutaminyl-tRNA synthetase was determined,and the results showed that the Glutaminyl-tRNA synthetase protein was successfully expressed in E. coli. The substrate glutamine did not inhibit the activity of glutaminyl-tRNA synthetase.

关 键 词：谷氨酰胺tRNA合成酶 原核表达 酶活性 

分 类 号：Q78[生物学—分子生物学]