PURCHASE OF FERMENTER AND OLIGONUCLEOTIDE SYNTHESIZER

发酵机和寡核苷酸合成仪的购买

• 批准号: 3519312
• 负责人: KATHLEEN S MATTHEWS
• 金额: $ 22万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-03-01 至 1988-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3519312
• 关键词: Clostridium; biotechnology; fermentation; microorganism; microorganism mass culture; microorganism metabolism; nucleic acid chemical synthesis; oligonucleotides; point mutation; synthetic nucleotide

Funds are requested for the purchase of a 250 liter fermenter and an oligonucleotide synthesizer. The primary application for these instruments will be in site-specific oligonucleotide-directed mutagenesis and isolation of the mutant proteins in large quantities. The instruments will be multi-user and will be available generally to the scientific community in Houston when use will not interfere with the projects of the investigators listed. In addition to the production of mutant proteins in large quantities, the fermenter will be utilized in several other projects which require large-scale fermentation (production of enzymes from Clostridia, production of bacteria for purification of wild-type proteins for routine use, and growth of bacteria for large-scale plasmid purification). The oligonucleotide synthesizer is required for the production of large quantities of specified sequences of DNA's; these will be used primarily for site-specific mutagenesis by standard methods using coding or recognition regions cloned in plasmids or in various Ml 3 vectors. A second application is the generation of synthetic oligonucleotides of various composition for use in binding studies in a variety of systems under investigation in the laboratories of the participating researchers. The coupling of these two instruments is essential for the generation of large quantities of selected mutant proteins; production of the mutant strains with site-specific oligonucleotides and the capacity to grow large quantities of cells will provide the resources to isolate sufficient amounts of the proteins for detailed chemical and physical characterization. Production of mutants for binding and repressor proteins, calmodulin, and promoter sequences, as well as metabolic studies of Clostridia strains, will be undertaken with these instruments. These important projects are entirely dependent on the availability of one or both of these instruments.

要求购买250升发酵罐和一个资金 寡核苷酸合成器。 这些乐器的主要应用 将在特定于位点的寡核苷酸指导的诱变和隔离 大量突变蛋白。 乐器将是 多用户，通常可用于科学界 休斯顿使用时不会干扰调查人员的项目 列出。 除了大型突变蛋白的产生 数量，发酵罐将用于其他几个项目 需要大规模发酵（从梭状芽胞杆菌生产酶， 生产细菌以纯化野生型蛋白质以进行常规 使用和细菌生长大规模质粒纯化）。 这 生产大型核苷酸合成器是必需的 DNA的指定序列的数量；这些将主要使用 使用编码或 克隆质粒或各种ML 3载体的识别区域。 一个 第二应用是生成的合成寡核苷酸 在各种系统中用于结合研究的各种组成 在参与研究人员的实验室进行调查。 这两种乐器的耦合对于产生至关重要 大量选定的突变蛋白；产生突变体 具有特异性寡核苷酸的菌株和大大生长的能力 数量的细胞将提供足够隔离的资源 用于详细化学和物理的蛋白质量 表征。 生产用于结合和阻遏物的突变体 蛋白质，钙调蛋白和启动子序列以及代谢研究 将使用这些工具进行梭状芽胞杆菌菌株。 这些 重要项目完全取决于一个或 这两种乐器。