SmPAP1-SmMYC复合体调控丹参酚酸类活性成分合成的机理研究

项目名称： SmPAP1-SmMYC复合体调控丹参酚酸类活性成分合成的机理研究

项目编号： No.31270338

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 王喆之

作者单位： 陕西师范大学

项目金额： 80万元

中文摘要： 酚酸类化合物是丹参重要的活性成分。我们发现转录因子SmPAP1及SmMYC与酚酸类成分的合成密切相关，SmPAP1对SmMYC的表达具有潜在的调控作用。推测丹参中存在SmPAP1-SmMYC复合体调控酶基因的表达进而控制酚酸类物质的合成，同时也调控SmMYC自身的表达，但其分子机制尚不清楚。本课题拟通过沉默SmMYC验证其在丹参酚酸类化合物合成中的关键作用；利用酵母双杂交和双分子荧光补偿法明确SmPAP1与SmMYC的相互作用；采用DNA酶I足迹、凝胶迁移率技术以及启动子瞬时激活体系研究SmPAP1对SmMYC启动子的特异结合及转录激活方式，并明确SmPAP1-SmMYC复合体作用于丹参酚酸类化合物合成途径的酶基因；进一步在丹参中进行SmPAP1及SmMYC的过表达或共表达。研究结果有望阐明SmPAP1-SmMYC调控丹参酚酸类活性成分合成的分子机制，为丹参高效、可行的改良提供思路及策略。

中文关键词： 酚酸类化合物；丹参；SmPAP1-SmMYC复合体；基因表达调控；

英文摘要： The phenolic acids are main active ingredients of S.miltiorrhiza. From our previous results, the expression of transcription factors SmPAP1 and SmMYC is correlated well with the accumulation of phenolic acids in S.miltiorrhiza, and SmPAP1 potentially regulates SmMYC expression. Accordingly, it is speculated that there may exist a regulation mode that SmPAP1-SmMYC complex regulate expression of the enzyme genes targeted and subsequently control phenolic acids biosynthesis, and activate SmMYC per si expression. However, the detailed interaction between the SmPAP1 and SmMYC and the mechanism for their subsequent coordinate regulation on phenolic acids biosynthesis still remain to be determined. In this proposal, we are planning to validate SmMYC as a key transcription factor governing phenolic acids biosynthesis of S.miltiorrhiza by SmMYC-specific silencing, and define the interaction between SmPAP1 and SmMYC using yeast two-hybrid combined with BiFC assay. Subequently, electrophoretic mobility shift assay (EMSA) and DNAase I footprinting will be employed to unravel the binding of SmPAP1 to specific region within SmMYC promoter. Besides, transcriptional activation of SmMYC itself and enzyme genes involved in phenolic acids biosynthesis under the control of SmPAP1-SmMYC complex will be investigated using transient

英文关键词： phenolic acids；S.miltiorrhiza；SmPAP1-SmMYC complex；gene expression regulation；