川泽泻鲨烯合酶基因的原核表达研究
发布时间：2019-07-06 21:17
【摘要】：目的:构建川泽泻鲨烯合酶(squalene synthase,SS)基因的原核表达载体。方法:在建泽泻SS基因序列的基础上,采用RT-PCR技术,从川泽泻新鲜叶片中克隆得到SS基因,并在BL21(DE3)细胞中分别用不同的表达载体、IPTG浓度、温度诱导该蛋白表达。结果:该基因在大肠杆菌中表达,但为包涵体,将其克隆到带有GST标签的pGEX-6t-1载体中进行原核表达并未改善蛋白溶解性;30℃诱导蛋白表达最佳;不同IPTG浓度对蛋白表达影响不大。结论:川泽泻的原核表达为后续对该基因的生物学功能及川泽泻品质改良奠定了基础。
文内图片：
图片说明：川泽泻总ＲNA凝胶电泳图
[Abstract]:Objective: to construct the prokaryotic expression vector of squalene polymerase (squalene synthase,SS) gene in Radix et Rhizoma Chuanze. Methods: on the basis of SS gene sequence, SS gene was cloned from fresh leaves of alisma orientalis by RT-PCR technique, and the expression of SS gene was induced by different expression vectors, IPTG concentration and temperature in BL21 (DE3) cells. Results: the gene was expressed in E. coli, but it was an inclusion body. Cloning it into pGEX-6t-1 vector with GST tag for prokaryotic expression did not improve the solubility of the protein. The best protein expression was induced at 30 鈩，
本文编号：2511332
文内图片：
图片说明：川泽泻总ＲNA凝胶电泳图
[Abstract]:Objective: to construct the prokaryotic expression vector of squalene polymerase (squalene synthase,SS) gene in Radix et Rhizoma Chuanze. Methods: on the basis of SS gene sequence, SS gene was cloned from fresh leaves of alisma orientalis by RT-PCR technique, and the expression of SS gene was induced by different expression vectors, IPTG concentration and temperature in BL21 (DE3) cells. Results: the gene was expressed in E. coli, but it was an inclusion body. Cloning it into pGEX-6t-1 vector with GST tag for prokaryotic expression did not improve the solubility of the protein. The best protein expression was induced at 30 鈩，
本文编号：2511332
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