MOLECULAR GENETICS OF POLYKETIDE BIOSYNTHESIS

描述(逐字摘自申请人的摘要)：甲藻是 聚酮的巨大生产者和一些最大和最复杂的 聚酮类化合物是由这些生物体产生的。最近，功能界别 聚酮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。这些策略包括： 组合表达，以及敲除或恢复活性的突变。这个 甲藻生物合成途径或偶联途径的功能表达 这些具有细菌系统的途径可能会导致前所未有的 工程生物合成新型聚酮化合物的生物合成能力。 该项目的第一个目标是聚酮的鉴定和作图。 利马原甲藻中的路径。为了实现这一目标，我们 必须采用能够在基因(S)中识别的策略 极大的基因组(1011个碱基)。虽然无疑是一个挑战，但初步的 数据表明，这并不是不可逾越的。第二个目标是发展 允许对通路进行功能识别的方法。 因为还没有开发出用于 甲藻，我们认为这项工作风险较高，但潜在的 福利是巨大的。冈田酸聚酮合成酶的鉴定 基因是迈向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 程序生物合成新化合物的基因。

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.