Core--Centralized DNA preparation, fermentation and extraction facility

对抗癌药物的需求仍然存在。新的天然产物的发现导致了几种可能被证明是有效抗癌药物的候选物的鉴定，其中包括从深水石质海绵中分离出来的海洋天然产物discodermolide。海洋微生物也提供了一种新的天然产物的来源，但对许多这些生物的次生代谢物表达知之甚少。这个项目的目标是在委内瑞拉链霉菌中表达这些微生物的基因。这将允许次级代谢物在一个明确的系统中表达，它也将允许组合生物学产生非自然的天然产物。这种潜在生物活性代谢物的新来源将与诺华的抗癌药物发现计划相结合，以确定新的抗癌药物。该项目包括许多功能，这些功能可以最有效地由单个Core设施处理。为此，HBOI的核心实验室将进行DNA提取、发酵和发酵液的提取，这是该项目的核心。微生物将从HBOI、俄勒冈大学和UCSC的研究项目中提供给核心实验室，用于DNA提取。纯化的DNA将被运往明尼苏达大学，在那里进行分子生物学研究。最终的克隆将被运回Core实验室，在那里进行小规模的发酵，提取，提取液将被运往诺华公司进行筛选。第一年的目标是20000条，随后的年份是30000条。在诺华筛选中显示活性的克隆将被参考以确认产生感兴趣的生物活性化合物。所有发酵生产用于生物测定指导的活性成分纯化的材料将由个别研究小组进行。

There continues to be a need for anti-cancer agents. The discovery of novel natural products has led to the identification of several candidates which may prove to be effective anti-cancer drugs, these include the marine natural product discodermolide which was isolated from a deep water lithistid sponge. Marine microorganisms are also providing to be a source of novel natural products however little is known about secondary metabolite expression in many of these organisms. It is the goal of this program project to express the genes from these microbes in Streptomyces venezuelae. This will allow the expression of secondary metabolite gens in a well defined system, it will also allow combinatorial biology to produce unnatural natural products. This novel source of potentially bioactive metabolites will be coupled with the Novartis anti-cancer drug discovery program in order to identify new anti-cancer agents. This project includes many functions which can most efficiently be handled by a single Core facility. To this end the core laboratory at HBOI will perform the DNA extractions, fermentations and extraction of fermentation broths which are central to this project. Microorganisms will be provided to the Core laboratory from the research projects at HBOI, the University of Oregon, and UCSC for DNA extraction. The purified DNA will be shipped to the University of Minnesota where the molecular biology will be performed. The resultant clones will be shipped back to the Core laboratory where they will be fermented at a small scale, extracted and the extracts shipped to Novartis for screening. The target for throughput is 20,000 clines in year one and 30,000 in subsequent years. Clones which show activity in the Novartis screens will be refermented to confirm the production of a bioactive compound of interest. All fermentations to produce material for bioassay-guided purification of the active component will be performed by the individual research groups.