FUNCTINAL DOMAINS OF BACCILLUS THURINGIENSIS ENDOTOXINS

苏云金芽孢杆菌内毒素的功能域

• 批准号: 2671982
• 负责人: DONALD H DEAN
• 金额: $ 36.28万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2671982
• 关键词: Lepidoptera; SDS polyacrylamide gel electrophoresis; affinity chromatography; aminoacid; aminopeptidase; bacterial proteins; bioassay; cytotoxicity; endotoxins; high performance liquid chromatography; insecticide biological effect; ion exchange chromatography; mass spectrometry; pest control; pesticide interaction; pesticide residues; protein structure function; receptor binding; site directed mutagenesis; tissue /cell culture; voltage /patch clamp

Bacillus thuringiensis is a microbial insecticide that is widely used to control numerous insects, including agricultural pests; and, mosquitoes and blackflies, vectors of human diseases. The active components are the insecticidal crystal proteins or Cry toxins. These insecticidal proteins may be manipulated by genetic and protein engineering to alter and improve their activity. The overall goal of this project is to investigate in detail the binding of several Cry toxins to a known receptor and improve the insecticidal activity by improving the binding to the receptor. The specific aims are to identify the amino acid residues on the Cry toxins that interact with the receptor and to experimentally alter these residues to identify which substituted residues enhance binding and toxicity. This will be accomplished by the genetic technique of site-directed mutagenesis. The components on the receptor that are responsible for interaction with the toxin will also be determined biochemically. The proposed research employs a model system consisting of a cry toxins of known structure as a well characterized receptor. When the basic understanding of toxin-receptor interaction is obtained, this may used to improve Cry toxins against the mosquito.

苏云金芽孢杆菌是一种微生物杀虫剂，广泛用于 控制多种昆虫，包括农业害虫；还有，蚊子 和黑蝇，人类疾病的媒介。 活性成分是 杀虫晶体蛋白或 Cry 毒素。 这些杀虫剂 蛋白质可以通过基因和蛋白质工程来操纵 改变和改善他们的活动。 该项目的总体目标是详细研究 多种Cry毒素作用于已知受体并提高杀虫效果 通过提高与受体的结合来发挥活性。 具体目标 是为了识别 Cry 毒素上相互作用的氨基酸残基 与受体并通过实验改变这些残基来识别 取代的残基增强了结合力和毒性。 这将是 通过定点突变的遗传技术来完成。 受体上负责相互作用的成分 毒素也将通过生化方法进行测定。 拟议的 研究采用了一个由已知的cry毒素组成的模型系统 结构作为一个明确表征的受体。 当基本 获得对毒素-受体相互作用的了解，这可以使用 改善Cry毒素对抗蚊子。