HIGH RESOLUTION PHYSICAL MAP OF CHROMOSOME 10

该项目的最终目标是对人类10号染色体进行测序。在 在目前的赠款期结束时，将提供一个基本的物理地图， 作为当前“STS内容”映射工作的结果而生成。这 地图将由1400个YAC克隆组成，这些克隆由300个STS和700个 杂交探针数学分析预测，这组YAC 将覆盖99.9%的染色体，并将在20个重叠群中排序， 每个重叠群的预期大小为7.7 Mb。这张地图也将是 与遗传学和细胞遗传学图谱相结合。 在续发补助金期间，有三个具体目标。一是 将验证STS内容映射。1400个YAC的集合， 地址已通过筛选确定了2层池，将殖民地 纯化，并重新筛选收集300 STS和700 杂交探针这一步将大大改善 1,000个锚本阶段的实物测绘产品将包含 两个部分：（1）每150 Kb一个锚密度的STS图;以及（2） 一组1400个对齐的YAC，将提供53的平均分辨率 KB. YAC将根据其地图位置排列， 分发给研究界。任何新的10号染色体STS将是 使用池化策略方便地映射到这个网格。 第二，将开发一个高分辨率的序列准备好的地图， 细菌人工染色体（BAC）作为模型。起初，高- 将使用VAC重叠群中的Alu间产物探测密度BAC网格 覆盖10p11.2的5 Mb区域。确定的200个BAC将 使用“克隆有限的”、非随机锚定策略进行比对，也称为 称为“无放置样本”。STS和杂交锚也将是 放置在BAC重叠群上，以便将BAC重叠群与 界标并比较BAC和YAC重叠群的STS含量。后 测试用例成功完成。将加大力度， 整个染色体。对齐的BAC的最大平均分辨率 13 KB。订购的BAC也将分发给研究 社区 第三，对6,000条10号染色体BAC的末端进行一次测序 将被启动。多重测序技术（Church和Kieffer- Higgens，1988）。这是一个大规模的测序项目 在CRI，将用于此目的。可开发更多STS 如果必要的话，从序列信息中提取。序列也将是 对数据库进行搜索以鉴定CpG岛，控制元件， 外显子、开放阅读框架和功能基序。这些映射的“序列 标签”，密度为每13 kb一个，将提供一个框架， 启动整个染色体的测序。在这次更新中， 期间，将为4，000个 “标签”

The ultimate goal of this project is to sequence human chromosome 10. At the end of the current grant period, a rudimentary physical map will be generated as a result of the current "STS content" mapping effort. This map will consist of 1400 YAC clones which are anchored by 300 STSs and 700 hybridization probes. Mathematical analyses predict that this set of YACs will cover 99.9% of the chromosome and will be ordered in 20 contigs with an expected size of 7.7 Mb per contig. This map will also have been integrated with the genetic and cytogenetic maps. For the renewal grant period, there are three specific aims. First, the STS content map will be validated. The collection of 1400 YACs, whose addresses have been identified by screening a 2-tier pool, will be colony purified, and re-screened with the collection of 300 STSs and 700 hybridization probes. This step will significantly improve the ordering of the 1,000 anchors. The physical mapping product at this stage will contain two parts: (1) an STS map at a density of one anchor per 150 Kb; and (2) a set of 1400 aligned YACs, which will provide an average resolution of 53 kb. The YACs will be arrayed according to their map positions and distributed to the research community. Any new chromosome 10 STSs will be conveniently mapped to this grid using a pooling strategy. Second, a high-resolution, sequence-ready map will be developed with bacterial artificial chromosomes (BACs) as the model. Initially, high- density BAC grids will be probed with inter-Alu products from a VAC contig covering a 5 Mb region from 10p11.2. The 200 identified BACs will be aligned using a "clone-limited", non-random anchoring strategy, also known as "sample-without-placement". STSs and hybridization anchors will also be placed on the BAC contigs in order to correlate the BAC contigs to landmarks and compare the STS content of the BAC and YAC contigs. After the test case is successfully completed. the effort will be scaled up for the entire chromosome. The maximum average resolution of the aligned BACs will be 13 kb. The ordered BACs will also be distributed to the research community. Third, one-pass sequencing of the ends of the 6,000 chromosome 10 BACs will be initiated. Multiplex sequencing techniques (Church and Kieffer- Higgens, 1988). which are being used for a large scale sequencing project at CRI, will be used for this purpose. Additional STSs can be developed from the sequence information if necessary. The sequences will also be searched against databases to identify CpG islands, control elements, exons, open reading frames and functional motifs. These mapped "sequence tags", at a density of one per 13 kb, will provide a framework when the sequencing of the entire chromosome is initiated. During this renewal period, approximately 1.6 Mb of raw sequences will be generated for 4,000 "tags".