A YEAST ARTIFICIAL CHROMOSOME CONTIG MAP OF CHROMOSOME 12

项目 2 的目标是开发一个高分辨率序列标记 基于 12 号染色体位点（STS）内容的图谱。我们提出了一个程序 与 Daniel Cohen 博士及其团队密切合作绘制了这张地图 巴黎 CEPH 的同事正在开发低分辨率物理 整个人类基因组图谱。 我们将获得12号染色体特异性 大插入酵母人工染色体 (YAC) 子文库 最近在巴黎建成的图书馆。 此外，科恩博士将 为我们提供有关 12 号染色体中 YAC 的信息 已通过指纹识别组装成重叠群的子文库 他们采用的方法。 我们将确定各个 YAC 的大小 这些重叠群中的每个 YAC 生成末端特异性序列。 所有确定源自 12 号染色体的末端序列 将被测序（至少部分地）以生成STS。 每个 YAC 将筛选重叠群内每个的存在或不存在 STS。 获得的二进制信息将用于构建STS- 基于内容的地图。 我们建议利用同源重组 构造极高分辨率地图的方法，其中 将推断出 STS 之间的精确顺序和距离。 我们提出一个 组合方法来识别差距并填补它们以完成 12号染色体的物理图。除了YAC端特有的STS之外， 我们将使用所有已使用的遗传标记 多个研究小组正在开发构建 12 号染色体的方法 物理图。 这个过程使我们能够整合遗传图谱 与我们将构建的物理地图一起在其他地方构建。 标记对之间的遗传距离可以与 我们将获得精确的物理距离来识别区域 高或低重组。 为了填补遗传图谱中的空白，我们 提出直接努力分离含有微卫星的序列 来自特定的 YAC 并使用它们来键入 CEPH 谱系。 我们的身体 绘图工作将补充并建立在 CEPH 的全球努力之上 和其他机构，可以为构建 人类个体染色体的高分辨率图谱。

The goal of Project 2 is to develop a high resolution sequence tagged site (STS) content based map of chromosome 12. We propose a program to build this map in close association with Dr. Daniel Cohen and his colleagues at CEPH in Paris who are developing a low resolution physical map for the entire human genome. We will obtain a chromosome 12 specific sublibrary of the large insert yeast artificial chromosome (YAC) libraries recently constructed in Paris. In addition, Dr. Cohen will provide us with information about those YACs in the chromosome 12 sublibrary which have been assembled into contigs by the fingerprinting methods they employ. We will ascertain the size of the individual YACs in these contigs generate end-specific sequences from each of the YACs. All end sequences which are determined to be derived from chromosome 12 will be sequenced, at least in part, to generate STSs. Each of the YACs within the contig will be screened for the presence or absence of each STSs. The binary information obtained will be used to construct an STS- content based map. We propose to utilize homologous recombination methods to construct an extremely high resolution map in which the precise order and distance between STSs will be deduced. We propose a combination of methods to identify gaps and to fill them to complete the physical map of chromosome 12. In addition to the YAC-end specific STS, we will use all of the genetic markers that have been utilized and that are being developed by several groups in constructing the chromosome 12 physical map. This procedure enables us to integrate the genetic maps being constructed elsewhere with the physical maps we will construct. The genetic distances between pairs of markers can be compared with precise physical distances that we will obtain to identify regions of high or low recombination. To fill in the gaps in the genetic map, we propose a directed effort to isolate microsatellite-containing sequences from specific YACs and use them to type the CEPH pedigrees. Our physical mapping effort will complement and build upon the global efforts at CEPH and other institutions and could provide a new paradigm for constructing high resolution maps of individual human chromosomes.