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。 在当前赠与期结束时，基本的实物地图将是 这是当前“STS内容”映射工作的结果。这 MAP将由1400个YAC克隆组成，分别由300个STS和700个STS锚定 杂交探针。数学分析预测，这组YAC 将覆盖99.9%的染色体，并将以20个重叠群进行排序 每个重叠群的预期大小为7.7 Mb。这张地图也将被 与遗传和细胞遗传学图谱相结合。 对于续签授权期，有三个具体目标。首先， STS内容地图将得到验证。1400个YAC的集合，他们的 通过筛选已确定地址的2层池，将其作为殖民地 提纯，并用收集的300个STS和700个STS重新筛选 杂交探针。这一步骤将显著改善 1000名主播。此阶段的物理测绘产品将包含 两部分：(1)密度为每150KB一个锚的STS映射；以及(2) 一组1400个对准的YAC，平均分辨率为53 KB。青年会将根据他们的地图位置和 分发给研究界。任何新的10号染色体STS都将是 使用池化策略方便地映射到此网格。 其次，将开发一张高分辨率的、随时准备好序列的地图 以细菌人工染色体(BAC)为模型。最初，高- 密度BAC网格将使用来自VAC重叠群的Alu间产品进行探测 覆盖从10p11.2开始的5Mb区域。已确定的200个BAC将是 使用“克隆受限”、非随机锚定策略对齐，也称为 为“不放置样本”。STSS和杂交锚点也将是 放置在BAC重叠群上，以便将BAC重叠群关联到 并比较BAC和YAC重叠群的STS含量。之后 测试用例已成功完成。这一努力将扩大到 整个染色体。对准BAC的最大平均分辨率 将是13kb。订购的BAC也将分发给研究人员 社区。 第三，6,000号染色体10号染色体末端的一次通过测序 将会被启动。多重测序技术(丘奇和基弗- 希金斯，1988)。它们正在用于一个大规模的测序项目 在CRI，将用于这一目的。可以开发更多的STSS 如有必要，从序列信息中删除。这些序列也将是 根据数据库进行搜索，以确定CpG岛、控制元素、 外显子、开放阅读框架和功能基序。这些被映射的“序列” 每13kb一个标签的密度将提供一个框架，当 开始对整个染色体进行测序。在此续订期间 期间，将为4,000个生成大约1.6Mb的原始序列 “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".