A YEAST ARTIFICIAL CHROMOSOME CONTIG MAP OF CHROMOSOME 12

项目2的目标是开发一个高分辨率的序列标签 基于站点(STS)内容的12号染色体图谱。我们提出了一个程序来 建立这张地图与丹尼尔·科恩博士和他的 巴黎CEPH的同事们正在开发一种低分辨率的物理 绘制整个人类基因组的图谱。我们将获得12号染色体特有的 大插入酵母人工染色体亚库(YAC) 最近在巴黎建造的图书馆。此外，科恩博士将 为我们提供有关12号染色体上的YAC的信息 通过指纹图谱组装成重叠群的亚文库 他们采用的方法。我们会确定个别青年会的规模。 在这些重叠群中，产生来自每个YAC的末端特异序列。 被确定为源自12号染色体的所有末端序列 将被测序，至少部分地，以产生STSS。每一届青年会 将对重叠群内的每一个存在或不存在进行筛选 STSS。所获得的二进制信息将被用于构建STS- 基于内容的地图。我们建议利用同源重组 方法来构建一个极高分辨率的地图，其中 将推导出STS之间的精确顺序和距离。我们提出了一个 识别差距并填补差距以完成 12号染色体的物理图谱。除了YAC末端特异的STS外， 我们将使用所有已被利用的遗传标记， 是由几个小组在构建12号染色体时开发出来的 物理地图。这一程序使我们能够整合基因图谱 正在用我们将要构建的物理地图在其他地方构建。 标记对之间的遗传距离可以与 我们将获得的精确物理距离，以识别 高或低的复合。为了填补基因图谱的空白，我们 提出一种定向分离微卫星序列的方法 从特定的YAC中提取，并用它们对CEPH家系进行分型。我们的体检 测绘工作将补充CEPH的全球努力并在此基础上再接再厉 和其他机构，并可以提供一个新的范式来构建 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.