BAC RESOURCE FOR CYTOGENETIC ANALSES OF HUMAN GENOME

描述：(申请人的描述)本提案的唯一目的是 确定3000个BAC，密度为每MBP约1个 人类基因组将作为原位和间接细胞遗传学分析的工具 硅胶。为了实现这一目标，我们将首先推出FISH-MAP&GT；3000 BAC 来自IRB批准的两个库。多数人将随机选出 来自其末端序列将被放置在辐射杂交体上的一组BAC 通过我们的合作者(亚当斯、文特尔、胡德和 考克斯)。通过对这些BAC进行FISH映射，我们将识别至少一个克隆 600条细胞遗传学条带中的每一条，或每5 Mbp一条。这些细胞遗传学标记 将因其在鱼类检测中的稳健和高效性能而被选中。 重要的是，这个过程将细胞遗传学和RH紧密结合在一起 地图。因此，选择BAC将是一项简单的练习 位于L-MBP间隔，从30,000将被放置在 未来3年内的RH地图。因此，所需的BAC组间隔为1 MBP和包含RH映射的STS，将在不扩展的情况下识别 图书馆屏幕。所有BAC都将是STC(BAC结束序列)的一部分 资源，这是一种有效地对基因组进行排序的策略的基础 经济实惠。因此，他们在最终序列中的位置 将准确地使研究人员容易地获得重叠 克隆和重排区域的序列。我们的建议包括计划 简化FISH程序，通过以下方式分发细胞遗传学定位数据 几个公共数据库，并分发两套细胞遗传学标记， 通过与Research的合作，每个频段1个集合和L每个MBP集合 遗传学。组装完成后，两套都将由FISH进行质量控制 对分池的分析。我们的战略还有一个额外的好处，那就是 将获得有关该基因的随机性和嵌合体频率的信息 这些文库是大规模测序工作的基础 人类基因组。此外，我们建议的随机鱼类调查是 足够的尺度来估计大尺度低气压的频率和分布 -复制复制，这是基因组的特征，很可能是 参与染色体重排的产生，与 基因家族的进化，以及完成序列的挑战 人类基因组的。

DESCRIPTION: (Applicant's Description) The single aim of this proposal is to identify 3000 BACs distributed at a density of ~ 1 per Mbp across the human genome to serve as tools for cytogenetic analyses in situ and in silico. To accomplish this aim, we will first FISH-map >3000 BACs derived from two IRB-approved libraries. The majority will be selected randomly from the set of BACs whose end-sequences will be placed on radiation hybrid maps through the efforts of our collaborators (Adams, Venter, Hood, and Cox). By FISH-mapping these BACs, we will identify at least one clone for each of 600 cytogenetic bands, or 1 per 5 Mbp. These cytogenetic markers will be selected for their robust and efficient performance in FISH assays. Importantly, this process will tightly integrate the cytogenetic and RH maps. As a consequence, it will be a simple exercise to select BACs situated at l-Mbp intervals from the 30,000 that will be positioned on the RH maps within the next 3 years. Thus, the desired set of BACs, spaced at 1 Mbp and containing RH-mapped STSs, will be identified without extensive library screens. All the BACs will be part of the STC (BAC end-sequence) resource, the basis of a strategy to sequence the genome efficiently and cost effectively. As a consequence, their positions in the final sequence will be known precisely allowing researchers to readily obtain overlapping clones and sequence of rearranged regions. Our proposal includes plans to streamline the FISH procedure, to distribute cytogenetic location-data via several public databases, and to distribute two sets of cytogenetic markers, a 1 -per-band set and a l-per-Mbp set, through a collaboration with Research Genetics. After assembly, both sets will be quality-controlled by FISH analyses of subpools. Our strategy also has the added benefit that critical information will be obtained on the randomness and chimerism frequency of the libraries that are the foundation of large-scale efforts to sequence the human genome. In addition, the random FISH survey we propose is of sufficient scale to estimate the frequency and distribution of large low -copy duplications, which are features of the genome that are likely to be involved in the generation of chromosomal rearrangements, associated with the evolution of gene families, and a challenge to completing the sequence of the human genome.