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丹参酮类化合物多属于二萜类化合物,是药用植物丹参的主要活性成分之一,在植物体内的生物合成需要多个基因的参与。CYP76AH1是丹参酮生物合成途径的第一个P450基因,前期的酵母表达以及体外酶促反应已经验证了其体外功能。为了对该基因的功能进行进一步研究,该文构建了该基因的丹参毛状根RNA干扰体系:通过Gateway技术构建能形成具有目标基因发卡结构的RNA干扰载体,转化发根农杆菌,转染丹参叶片,将干扰片段整合至丹参基因组中,获得了高效干扰的CYP76AH1干扰型丹参毛状根。
Tanshinones are mostly diterpenoids and are one of the main active components of medicinal plant Salvia miltiorrhiza. Biosynthesis in plants requires the involvement of multiple genes. CYP76AH1 is the first P450 gene of tanshinone biosynthesis pathway. Previously, yeast expression and in vitro enzymatic reaction have verified its in vitro function. In order to further study the function of this gene, this paper constructed the RNA interference system of Salvia miltiorrhiza hairy root: constructing RNA interference vector with the hairpin structure of target gene by Gateway technology, transforming Agrobacterium rhizogenes, transfection of Salvia miltiorrhiza Leaves, the interference fragment was integrated into the Salvia genome, obtained efficient interference CYP76AH1 interference Salvia hairy roots.