Evolution of Pathways to Pharmaceuticals in the Poppy Family

描述（由申请人提供）：目前大约25%的药物要么直接从自然界获得，要么在结构上基于天然产物，包括来自植物的重要抗癌药物和止痛药。从植物中选择药用化合物的研究需要对包含药物的整个生物合成途径的植物物种进行详细的调查。关于这些高度专门化的植物的详细的遗传和生化信息常常是缺失的。为了填补这一知识空白，并推进罂粟科植物衍生药物的发现和生产领域的研究，我们建议利用EST、454 FLX和Solexa测序组合，通过比较转录组学来鉴定罂粟罂粟（Papaver somniferum）和加州罂粟（Eschscholzia californica）中生物碱谱不同的植物器官中表达的基因。将为每个物种开发代表转录组的微阵列，以便确定所有三种罂粟物种和somniferum转基因植物正常发育期间的时间和空间表达模式，其中六种已知生物碱合成基因系统地过表达或过表达。将准备BAC文库，以便构建这两种罂粟科成员生物碱生物合成基因周围的基因组区域的物理图谱。序列数据的分子进化分析将在基因家族系统发育、基因表达模式和生物碱含量的背景下进行，以确定可能编码参与生物碱生物合成的酶或转录因子的新基因。功能分析将与基因进行假设编码酶参与生物合成药用生物碱。这些遗传和生化信息将以用户友好的数据库（PoppyDB）和查看器（PhyloPathway）的形式公开提供，用于代谢工程、仿生合成和定向进化，以改善药物生产和开发新药。

DESCRIPTION (provided by applicant): Approximately 25% of current pharmaceuticals are either directly obtained from nature or are structurally based upon natural products that include important anti-cancer drugs and painkillers from plants. The study of selected medicinal compounds from plants requires detailed investigation of exactly that plant species that contains the entire biosynthetic pathway to the drug. Detailed genetic and biochemical information on these highly specialized plants is often missing. It is proposed to fill this knowledge gap and advance research in the field of plant-derived pharmaceuticals discovery and production in the poppy family (Papaveraceae) by identifying genes expressed in plant organs distinct in their alkaloid spectrum in opium poppy Papaver somniferum and the California poppy Eschscholzia californica through comparative transcriptomics using a combination of EST, 454 FLX and Solexa sequencing. Microarrays that represent the transcriptome will be developed for each species so that the temporal and spatial expression patterns during normal development in all three poppy species and in transgenic plants of P. somniferum in which six of the known alkaloid synthesis genes are systematically over- or under-expressed can be determined. BAC libraries will be prepared so that physical maps can be constructed of genomic regions surrounding alkaloid biosynthesis genes in these two members of the poppy family. Molecular evolutionary analysis of sequence data will be carried out within the context of gene family phylogenies, gene expression patterns and alkaloid content in order to identify novel genes that may encode enzymes or transcription factors involved in alkaloid biosynthesis. Functional analyses will be performed with the genes hypothesized to encode enzymes involved in the biosynthesis of medicinal alkaloids. This genetic and biochemical information will be made publicly available in the form of a user-friendly database (PoppyDB) and viewer (PhyloPathway) for metabolic engineering, biomimetic synthesis and directed evolution for the improved production of drugs and for the development of novel drugs. Project Narrative: Approximately 25% of current pharmaceuticals are either directly obtained from nature or are structurally based upon natural products that include important anti-cancer drugs and painkillers from plants. The study of selected medicinal compounds from plants requires detailed investigation of exactly that plant species that contains the synthetic pathway to the drug. Detailed genetic and biochemical information on these highly specialized plants is often missing; it is proposed to fill this knowledge gap and advance research in the field of plant-derived pharmaceuticals discovery and production in the poppy family.