VERTICALLY INTEGRATED YAC BASED X CHROMOSOME ANALYSIS

基于YAC的垂直整合X染色体分析：第一次 人类基因组计划的第一阶段，这里的人类基因组中心 提供了实现第一代地图的思想、方法和数据 7号染色体和X染色体。 这些地图是基于重叠的酵母 人工染色体（YAC），并与序列标记格式化 通过聚合酶链反应（PCR）检测的位点（STS）。 该中心的下一个目标是实现一个扩展的，垂直整合的 分析顺利进行，一直到测序。 作为第一 步骤，X染色体图谱将至少用STS标记进行细化 每100 kb，并将与基因和标记， 多态性（项目1）。 这将提供一个平台， 研究人员寻找疾病基因或研究结构或发育 生物学，也将作为一个起始框架， 进行高通量测序 数据将被组织起来，并与外部来源的数据相结合， 一个信息处理和分析核心，将提交合理化的 及时将地图和基础数据输入国际数据库， 时尚. 材料，包括克隆文库和STS引物对，将 在映射资源和实现核心中存储和维护， 这也将有助于调整方法，使之适用于一般用途， 来自项目实验室和外部来源的协议。 更精细的绘图和测序所需的新技术将是 开发，包括， 在项目1中，建立更稳定的新青年中心;发展和 改进寻找基因的方法，包括新的重组类型 捕获由YAC编码的cDNA;使用简并STS来寻找 基因家族成员的位置;以及基于PCR的适应性 从短序列开始筛选全长cDNA; 在项目2中，测试有序鸟枪测序方法， 最佳可用替代品;以及 此外，比较基因组分析将有助于测序 在小鼠和人类DNA中的选定区域， 通过向研究小组提供保守的STS， 研究染色体结构和内容的进化。

Vertically Integrated YAC-Based X Chromosome Analysis: In the first phase of the human genome initiative, the Human Genome Center here provided the ideas, methods, and data to achieve first-generation maps of chromosomes 7 and X. The maps are based on overlapping yeast artificial chromosomes (YACs), and are formatted with sequence-tagged sites (STSs) that are detected by the polymerase chain reaction (PCR). The Center's next goal is to achieve an extended, vertically integrated analysis that proceeds smoothly all the way to sequence. As a first step, the X chromosome map will be refined with STS markers at least every 100 kb, and will be annotated with genes and markers that detect polymorphism (Project 1). This will provide a platform for the needs of researchers seeking disease genes or studying structural or developmental biology, and will also serve as a starting framework to merge mapping with high-throughput sequencing. Data will be organized and integrated with data from outside sources in an Information Handling and Analysis Core, which will submit rationalized maps and the underlying data to international databases in a timely fashion. Materials, including clone libraries and STS primer pairs, will be stored and maintained in a Mapping Resources and Implementation Core, which will also aid in the adaptation for general use of methods and protocols coming from Project laboratories and outside sources. New technology required for the finer mapping and sequencing will be developed, including, in Project 1, the institution of more stable new YACs; development and adaptation of methods to find genes, including new types of recombination trapping of cDNAs encoded by YACs; use of degenerate STSs to find the locations of members of gene families; and adaptation of PCR-based screening to find full-length cDNAs starting from short sequences; in Project 2, testing an ordered shotgun approach to sequencing compared to the best available alternatives; and In addition, comparative genome analysis would be aided by the sequencing of selected regions in mouse and human DNA that are syntenically equivalent, and by the provision of conserved STSs to research groups studying the evolution of chromosome structure and content.