STRUCTURAL STUDIES OF HUMAN CHROMOSOME 11

低分辨率，基于YAC的人类基因组部分图谱，包括 几个完整的人类染色体，在过去已经开发出来， 几年 然而，用于创建详细， 显示重要层序位置的高分辨率地图 结构和位置，方向和转录框架 基因还没有被开发出来。 本申请提出了一种有限的 可行性研究，以测试开发高分辨率DNA的概念， 基于新策略的人类染色体序列图 称为GSS（基因组序列样本）映射。 这一战略取决于 从染色体特异性文库中鉴定粘粒克隆 通过与来自最小平铺路径染色体的YAC克隆杂交 图，通过单个克隆开发高深度粘粒重叠群 指纹分析和粘粒克隆的直接末端测序， 生成一组有序序列片段。 这张高分辨率地图 将允许识别基因序列和序列基序， 基于计算机的搜索算法，编写基本 基因组任何区域的连续STS标记，并提供模板 最终完成人类染色体DNA测序的材料。 该项目的最初重点将是开发一个高 分辨率图的人类染色体11，但将包括其他地区的 在时间允许的情况下，研究生物学意义上的基因组。 该项目将 还开发高吞吐量，平衡的自动化系统， 兆碱基DNA测序和高分辨率物理作图。

Low resolution, YAC-based maps of portions of the human genome, including several complete human chromosomes, have been developed over the past several years. However, the techniques for the creation of detailed, high resolution maps showing the locations of important sequence structures and the locations, orientation and transcription frame of genes, have not yet been developed. This application proposes a limited feasibility study to test the concept of developing high resolution, DNA sequence-based maps of human chromosomes based upon a novel strategy called GSS (Genome Sequence Sample) mapping. This strategy depends upon the identification of cosmid clones from chromosome-specific libraries by hybridization with YAC clones from a minimum tiling path chromosome map, the development of high depth cosmid contigs by individual clone fingerprinting, and the direct end-sequencing of cosmid clones to generate a set of ordered sequence fragments. This high resolution map will allow the identification of gene sequences and sequence motifs by computer-based search algorithms, the preparation of essentially continuous STS markers for any region of the genome, and provide template material for the eventual complete DNA sequencing of human chromosomes. The initial focus of this project will be the development of a high resolution map of human chromosome 11 but will include other regions of the genome of biological interest as time permits. This project will also develop high throughput, balanced automation systems for the megabase DNA sequencing and for high resolution physical mapping.