LASER RAMAN PROBE OF MOLECULAR STRUCTURE IN LIVING CELLS

活细胞分子结构的激光拉曼探针

• 批准号: 3283895
• 负责人: WARNER L PETICOLAS
• 金额: $ 4.49万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-08-01 至 1987-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3283895
• 关键词: DNA; Raman spectrometry; antibody; chromosomes; conformation; lasers; nucleic acid structure; plants; respiratory epithelium; salivary glands; ultraviolet microscopy

It is proposed to build a laser Raman microscope which is capable of observing chromosomal material with about one micron resolution. Using this apparatus Raman spectra will be taken of chromosomes from a variety of easily available sources. By focusing a laser beam of about 1 micron or less on various portions of the chromosome and associated cellular apparatus and capturing the Raman light which is back scattered through the microscope objective, it is proposed to obtain both the classical and the resonance Raman spectra of the various chromosomal components including both protein and DNA. In particular the Raman marker band characteristic of the A, B, C, and Z forms of DNA as well as Raman bands sensitive to protein secondary structure will be sought. The presence of the Z marker bands in a portions of a gene would give evidence for the Z form. Polytene chromosomes will be examined in fixed and non-fixed states. The Raman spectra of polytene chromosomes containing bound antibodies to Z-DNA will also be taken to see if evidence can be found for presence of the Z-conformation in the DNA of polytene chromosomes upon antibody binding. This approach could settle the questions of whether or not Z-DNA exists in polytene chromosomes or whether or not antibodies are capable of pulling the DNA into the Z form upon binding. By collecting instanteous Raman spectra of the cellular components of living cells, it is proposed to determine the type, extent of and duration of conformational changes in the DNA and associated proteins during cell division. Thus, it is hoped to answer the questions as to whether Z-DNA is a naturallly occurring structure in genetic material. It is also hoped to improve the understanding of the molecular basis of the hereditary process. Such understanding can only help to formulate a more rational approach to the development of drug therapy for treatment of diseases which involve malfunction of the genetic material.

有人建议建造一台激光拉曼显微镜，它能够 以大约一微米的分辨率观察染色体材料。vbl.使用 这台仪器将对来自不同物种的染色体进行拉曼光谱测量 容易获得的资源。通过聚焦约1微米的激光束或 在染色体和相关细胞的不同部分上较少 设备，并捕获后向散射通过 显微镜物镜，提出既能获得经典物镜，又能获得 不同染色体组分的共振拉曼光谱 蛋白质和DNA都是。尤其是拉曼标记带特性 DNA的A、B、C和Z形式以及对 将寻找蛋白质的二级结构。Z标记的存在 基因部分中的条带将为Z型提供证据。多烯 染色体将在固定和非固定状态下进行检查。拉曼 含有Z-DNA Will结合抗体的多线染色体的光谱 也要看看是否能找到证据证明 抗体结合时多线染色体DNA中的Z构象。 这一方法可以解决是否存在Z-DNA的问题 多线染色体或抗体是否能够拉动 DNA在结合时变成Z形。通过收集即时拉曼 活细胞的细胞成分的光谱，建议 确定构象变化的类型、程度和持续时间 细胞分裂过程中的DNA和相关蛋白质。因此，希望能够 回答关于Z-DNA是否是自然发生的问题 遗传物质中的结构。也希望能改善 对遗传过程的分子基础的理解。是这样的 理解只会有助于制定一种更理性的方法来处理 药物治疗包括以下几种疾病的发展 遗传物质的故障。