Yi Yi, Qiao Dairong, Bai Linhan, Xu Hui, Li Ya, Wang Xiaolin, Cao Yi
Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, College of Life Sciences, Sichuan University, Sichuan 610064, P. R. China.
J Microbiol. 2007 Apr;45(2):153-7.
5-enolpyruvylshikimate-3-phosphate synthase (EPSP synthase, EC 2.5.1.19) is the sixth enzyme in the shikimate pathway which is essential for the synthesis of aromatic amino acids and many secondary metabolites. The enzyme is widely involved in glyphosate tolerant transgenic plants because it is the primary target of the nonselective herbicide glyphosate. In this study, the Dunaliella salina EPSP synthase gene was cloned by RT-PCR approach. It contains an open reading frame encoding a protein of 514 amino acids with a calculated molecular weight of 54.6 KDa. The derived amino acid sequence showed high homology with other EPSP synthases. The Dunaliella salina EPSP synthase gene was expressed in Escherichia coli and the recombinant EPSP synthase were identified by functional complementation assay.
5-烯醇丙酮酰莽草酸-3-磷酸合酶（EPSP合酶，EC 2.5.1.19）是莽草酸途径中的第六种酶，该途径对于芳香族氨基酸和许多次生代谢产物的合成至关重要。该酶广泛存在于耐草甘膦转基因植物中，因为它是非选择性除草剂草甘膦的主要作用靶点。在本研究中，通过RT-PCR方法克隆了盐生杜氏藻EPSP合酶基因。它包含一个开放阅读框，编码一个由514个氨基酸组成的蛋白质，计算分子量为54.6 kDa。推导的氨基酸序列与其他EPSP合酶具有高度同源性。盐生杜氏藻EPSP合酶基因在大肠杆菌中表达，并通过功能互补试验鉴定了重组EPSP合酶。
