MOLECULAR GENETICS OF POLYKETIDE BIOSYNTHESIS

描述（来自申请人摘要的逐字记录）：甲藻是 聚酮化合物的巨大生产商和一些最大最复杂的 聚酮化合物由这些生物体产生。最近，功能 聚酮化合物生物合成途径的表达导致了几种策略 用于生产稀有或新颖的聚酮化合物。这些战略包括， 组合表达和敲除或恢复活性的突变。的 甲藻生物合成途径的功能表达或偶联 这些途径与细菌系统的结合可能会导致前所未有的 生物合成能力，用于新型聚酮化合物的工程化生物合成。 本项目的第一个目标是聚酮化合物的鉴定和作图 在甲藻原甲藻利马途径。为了实现这一目标，我们 必须采用策略，将允许在一个或多个基因的识别， 非常大的基因组（1011 bp）。虽然毫无疑问是一个挑战， 数据表明，这并非不可克服。第二个目标是发展 方法，将允许的途径的功能鉴定。 因为没有遗传转化系统已经开发出来， 甲藻，我们认为这项工作有点高风险，但潜在的 福利很好。冈田酸聚酮合酶的鉴定 基因是我们实现克隆和测序这一长期目标的第一步 整个生物合成途径，并利用甲藻生物合成 新化合物的程序性生物合成基因。

DESCRIPTION (verbatim from the applicant's abstract): Dinoflagellates are prodigious producers of polyketides and some of the largest and most complex polyketides are produces by these organisms. Recently, the functional expression of polyketide biosynthetic pathways has led to several strategies for producing rare or novel polyketides. These strategies include, combinatorial expression, and mutations that knockout or restore activity. The functional expression of dinoflagellate biosynthetic pathways or the coupling of these pathways with bacterial systems could lead to unprecedented biosynthetic capabilities for the engineered biosynthesis of novel polyketides. The first goal of this project is the identification and mapping of polyketide pathways in the dinoflagellate Prorocentrum lima. To accomplish this goal, we must employ strategies that will permit the identification of the gene(s) in an extremely large genome (1011 bp). While undoubtedly a challenge, preliminary data indicate that it is not insurmountable. The second goal is to develop methods that would permit the functional identification of the pathways. Because no genetic transformation systems have been developed for dinoflagellates, we view this work as somewhat high risk, however the potential benefits are great. The identification of the okadaic acid polyketide synthase gene is the first step towards our longer term goal of cloning and sequencing the entire biosynthetic pathway, and utilizing dinoflagellate biosynthetic genes for programmed biosynthesis of novel compounds.