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: 4689017
• 负责人: R W HARTLEY
• 金额: --
• 依托单位: NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & 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/4689017
• 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 and 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 barnase gene carrying an inactivating transposon and a fragment of the wild type gene overlapping the transposon insertion site have been cloned in E. coli plasmids. Reconstructions demonstrate that the native gene is lethal in E. coli. The entire gene has been sequenced, confirming the amino acid sequence and revealing a signal sequence typical of exported proteins. The crippled gene is transcribed in both E. coli and B. subtilis.

两种蛋白质，藤蔓酶，芽孢杆菌胞外核糖核酸酶