BAC RESOURCE FOR CYTOGENIC ANALYSES OF HUMAN GENOME

描述：（申请人的描述）本提案的唯一目的是 以识别3000个BAC，这些BAC以每Mbp约1个的密度分布在 人类基因组作为细胞遗传学分析的工具， silico。 为了实现这一目标，我们将首先FISH地图>3000 BAC派生 两个经IRB批准的图书馆 大多数人将被随机挑选 从BAC的集合中，其末端序列将被放置在辐射混合物上， 通过我们的合作者（亚当斯，文特尔，胡德， 考克斯）。 通过对这些BAC进行FISH定位，我们将鉴定出至少一个克隆， 每600条细胞遗传学带，或每5 Mbp 1条。这些细胞遗传学标记 将选择其在FISH测定中的稳健和有效性能。 重要的是，这一过程将紧密整合细胞遗传学和RH 地图 因此，选择BAC将是一项简单的工作 位于距离30，000的l-Mbp间隔处， 未来三年内的RH地图。 因此，期望的BAC集合，间隔为1 Mbp和含有RH映射的STS，将在没有广泛的 图书馆屏幕 所有BAC将成为STC（BAC末端序列）的一部分 资源，一个战略的基础，以测序基因组有效和 成本效益。 因此，它们在最终序列中的位置 将被精确地知道，使研究人员能够容易地获得重叠的 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位置数据 几个公共数据库，并分发两套细胞遗传学标记， 通过与研究中心的合作， 遗传学 组装后，两套将由FISH进行质量控制 子池分析。 我们的战略还有一个额外的好处， 将获得关于随机性和嵌合频率的信息， 这些库是大规模测序的基础， 人类基因组 此外，我们建议的随机FISH调查是 有足够的尺度来估计大规模低 - 复制，这是基因组的特征，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。

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.