三七中ss基因超表达载体的构建及其遗传转化the-药学学报.pdfVIP

138 药学学报 Acta Pharmaceutica Sinica 2013, 48 ( 1) : 138 143 三七中SS 基因超表达载体的构建及其遗传转化 * 颖, 赵恒伟, 葛 锋 , 石 磊, 刘迪秋 ( 昆明理工大学生命科学与技术学院, 云南 昆明 650500) 摘要: 三七总皂苷为三七主要的药用活性成分, 但因其产量较低, 使其药用价值很难得到推广。随着皂苷类 物质生物合成途径研究的不断深入, 通过代谢工程手段调控三七皂苷的生物合成逐步成为可能。本研究构建了 皂苷合成途径中鲨烯合酶基因的超表达载体, 并利用农杆菌介导法将超表达载体转移并整合至三七基因组中, 通过 PCR 检测及皂苷含量检测, 初步明确鲨烯合酶基因在三七皂苷生物合成途径中的作用, 为将来三七总皂苷 同源或异源高效表达系统的构建提供理论依据和技术基础。 关键词: 三七; 愈伤组织; 三七总皂苷; 单体皂苷; 鲨烯合酶 中图分类号: R931 文献标识码:A 文章编号: 05 13-4870 (2013) 01-0138-06 The construction of over-expression vector for Panax notoginseng SS gene and its transformation * SUN Ying, ZHAO Heng-wei, GE Feng , SHI Lei, LIU Di-qiu (Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China) Abstract: PNS (Panax notoginseng saponins) is the main medical bioactive component in Panax notoginseng. The medical value of PN S cannot be extended because of its low production. With the deep study of saponins biosynthetic pathway, the control of PNS biosynthesis through metabolic engineering has gradually become possible. In this study, the Squalene sy nthase (SS) over-expression vector was established. By the way of agrobacterium-mediated method, the vector was transfered and integrated into the Panax notoginseng genome. The result of the PCR detection and the saponin content detection shows that over-expression SS is able to produce high level of