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。
PNS(Panax notoginseng saponins) is the main medical bioactive component in Panax notoginseng.The medical value of PNS 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 synthase(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 Panax notoginseng saponins,and confirms the regulatory function of SS in the biosynthesis of ginsenosides in Panax notoginseng.It provides a theoretical basis and technical basis for the construction of PNS homologous or heterologous efficient expression system in the future.