ADVANCED QUANTITATIVE IMAGING FOR DNA PHYSICAL MAPPING

这个多学科项目的广泛目标是推进遗传学 通过开发基于计算机的系统进行大规模DNA绘图的研究 1）加快DNA大区域的物理映射， 单个酵母染色体的规模;以及2）改善大规模人类染色体的遗传学特性。 连锁作图 待开发的领域是先进的形象- 用于提取信息的采集和处理系统 通过平板凝胶分离的核酸分子的分级混合物 电泳 具体目标是：1）开发一个直接（非摄影） 用荧光DNA结合染料染色的凝胶成像; 2）延长 系统，以适应用于RFLP作图的放射自显影图; 3）开发 用于自动高分辨率检测的定量成像算法 用于片段大小测定的凝胶带的强度估计 从能带迁移率和数据完整性评价; 4）应用方法 用于凝胶图像的几何失真补偿， 通过不同方法获得的图像的分析比较， 5）开发一个交互式软件系统， 数字化凝胶的快速有效的专家解释和分析 图像在一个国家的最先进的工作站环境;和6）评估 系统的有效性，以满足遗传研究的需要。 方法包括使用先进的CCD成像设备， 摄影限制;应用本地开发的图像估计 算法，以改善定量分析的DNA消化， 限制酶;使用“扭曲”算法来补偿 电泳畸变;执行接收器操作特性 (REC)和评估数据库研究，以评估检测算法 性能;提供用于增强凝胶图像的图像处理实用程序 特征;以及通过以下方式辅助凝胶图像的交互式解释： 基于可接受的窗口化和联网来构造用户界面 标准，以确保软件的可移植性和方便未来的出口。 这三方合作涉及遗传学系， 计算机科学系（CS）和生物医学计算机 实验室，都在华盛顿大学。 独立资助的CS 组件正在开发一种DNA分析系统， 分析大量的DNA片段大小数据和凝胶图像数据 使用这里提出的系统生成。

The broad goal of this multi-disciplinary project is to advance genetics research in large-scale DNA mapping by developing a computer-based system for: 1) expediting the physical mapping of large regions of DNA on the scale of individual yeast chromosomes; and 2) improving large-scale human linkage mapping. To be developed and fields is an advanced image- acquisition and processing system for extracting information from fractionated mixtures of nucleic-acid molecules separated by slab-gel electrophoresis. Specific aims are to: 1) develop a system for direct (non-photographic) imaging of gels stained with fluorescent DNA-binding dyes; 2) extend the system to accommodate autoradio-grams used in RFLP mapping; 3) develop quantitative-imaging algorithms for automated, high-resolution detection and intensity estimation of gel bands for fragment-size determination from band mobilities and for data-integrity evaluation; 4) apply methods for geometric-distortion compensation of gel images to allow direct analytic comparisons of images obtained by different methods and from different laboratories; 5) develop an interactive software system for rapid and efficient expert interpretation and analysis of digitized gel images in a state-of-the-art workstation environment; and 6) evaluate the system's effectiveness in meeting the needs of the genetic research. Methods include using an advanced CCD imaging device to avoid photographic limitations; applying locally developed image-estimation algorithms to improve quantitative analysis of DNA digested by restriction enzymes; using "warping" algorithms to compensate for electrophoretic aberrancies; performing receiver-operating-characteristic (REC) and evaluation-database studies to assess detection-algorithm performance; providing image-processing utilities for enhancing gel-image features; and aiding interactive interpretation of gel images by constructing a user interface based on accepted windowing and networking standards to ensure software portability and ease of future exportation. This three-way collaboration involves the Department of Genetics, the Department of Computer Science (CS), and the Biomedical Computer Laboratory, all at Washington University. The independently funded CS components is developing a DNA analysis system for maintaining and analyzing large amounts of both DNA-fragment-size data and gel-image data to be generated using the system here proposed.