PURCHASE OF FERMENTER AND OLIGONUCLEOTIDE SYNTHESIZER

请拨资金购买一个250升发酵罐和一个 寡核苷酸合成器。这些工具的主要应用 将在位点特异的寡核苷酸定向突变和分离 大量的突变蛋白质。这些仪器将是 多用户，并将在#年向科学界普遍提供 休斯顿使用时不会干扰调查人员的项目 已列出。除了大量生产突变蛋白外 数量，发酵罐将用于其他几个项目，这些项目 需要大规模发酵(从梭状芽胞杆菌产生酶， 常规野生型蛋白提纯菌种的生产 用于大规模纯化的细菌的使用和生长)。这个 大分子大分子的生产需要使用寡核苷酸合成器 指定DNA序列的数量；这些将主要用于 对于通过使用编码或 识别区域克隆在质粒或不同的mL3载体中。一个 第二个应用是合成合成的寡核苷酸 在各种体系中用于结合研究的各种组合物 正在参与研究人员的实验室进行调查。 这两个工具的耦合对于产生 大量选定的突变蛋白；突变蛋白的生产 具有位点特异性寡核苷酸和生长能力的菌株 大量的细胞将提供分离足够的资源 详细的化学和物理蛋白质的数量 人物刻画。结合和阻遏因子突变体的生产 蛋白质、钙调蛋白和启动子序列以及代谢研究 将使用这些仪器进行梭状芽胞杆菌菌株的研究。这些 重要的项目完全依赖于一个或多个 这两种乐器都是。

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.