生物学杂志

生物学杂志

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谷氨酰胺tRNA合成酶的克隆表达纯化及活性测定

  

  1. 1苏州大学基础医学与生物科学学院，苏州，215123
  • 出版日期:2016-02-18 发布日期:2016-02-18
  • 基金资助:
    国家自然科学基金项目（No. 81071306）

Moleculor Cloning and Expession and Purification of Glutaminyl-tRNA Synthetase

  • Online:2016-02-18 Published:2016-02-18

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摘要：  

为了研究谷氨酰胺tRNA合成酶结构和性质，人工合成谷氨酰胺tRNA合成酶基因，构建原核表达载体pPET-28a-His-GlnRS和pPIDTpSMART-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。

关键词: 谷氨酰胺tRNA合成酶,  , 原核表达,  , 酶活性

Abstract:  In order to study the structure and function of Glutaminyl-tRNA Synthetase,we synthesise the Glutaminyl-tRNA Synthetase and -tRNAGlngene, then the gene was subcloned into expression vector PET-28a-His-GlnRS and PIDTpSMART,separately , expessed Glutaminyl-tRNA Synthetase protein in BL21(DE3) by IPTG induction and we determined the activity of purify Glutaminyl-tRNA Synthetase Results showed that the Glutaminyl-tRNA Synthetase protein had been successfully expressed in E.coli. The substrate glutamine do not inhibit the activity of Glutaminyl-tRNA Synthetase.

Key words: Glutaminyl-tRNA Synthetase; Prokaryotic expression, Enzyme activity