RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUENFACIENS

解淀粉芽孢杆菌的核糖核酸及其抑制剂

• 批准号: 3940232
• 负责人: R W HARTLEY
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3940232
• 关键词: Bacillus; Bacillus subtilis; Escherichia coli; X ray crystallography; affinity chromatography; bacterial genetics; circular dichroism; conformation; enzyme complex; enzyme model; enzyme structure; fluorescence spectrometry; gel filtration chromatography; gene expression; genetic manipulation; genetic recombination; genetic regulation; genetic transcription; ion exchange chromatography; molecular cloning; nucleic acid sequence; pancreatic ribonuclease; structural genes; thermodynamics; tissue /cell culture; transposon /insertion element; ultraviolet spectrometry

Two proteins, barnase, the extracellular ribonuclease of Bacillus amyloliquefaciens, and barstar, its intracellular inhibitor, are used as a model system for the study of protein folding a protein- protein interactions. Barnase is one of an homologous group of ribonucleases occurring in both prokaryotes and eukaryotes. Recombinant DNA techniques are being applied to the project with three major aims: 1) to facilitate production, 2) to examine the structural and control sequences of the genes and 3) to tailor specifically designed modifications in the sequences to test theories of protein folding. The lethal effect of the cloned wild-type barnase gene in either E. coli or B. subtilis can be repressed by expression of the cloned barstar gene placed on the same plasmid. Secretion vectors for both proteins have been devised. Both genes have been sequenced, providing confirmation of the barnase sequence (110 residues) and a derived sequence (89 residues) for barstar which agrees well with previous amino-acid analysis. Effects of several site-directed mutations of barnase on activity and thermal stability have been studied.

两种蛋白质，芽孢杆菌RNA酶，芽孢杆菌的胞外核糖核酸酶 解淀粉菌及其细胞内抑制剂芽孢杆菌RNA酶抑制剂 作为研究蛋白质折叠的模型系统 蛋白质相互作用 芽孢杆菌RNA酶是一种同源的 在原核生物和真核生物中都存在的核糖核酸酶。 重组DNA技术正在应用于该项目 有三个主要目标：1）促进生产，2）检查 基因的结构和控制序列，以及3）定制 在序列中专门设计的修饰以测试 蛋白质折叠理论 克隆的野生型芽孢杆菌RNA酶基因在大肠杆菌中的致死作用。 coli或B.枯草芽孢杆菌可以通过表达克隆的 将芽孢杆菌RNA酶抑制剂基因置于同一质粒上。 分泌载体 这两种蛋白质都被设计出来了。 这两个基因都已测序，证实了 芽孢杆菌RNA酶序列（110个残基）和衍生序列（89 残基），其与先前的氨基酸一致 分析. 芽孢杆菌RNA酶几种定点突变对活性的影响 和热稳定性进行了研究。