A YEAST ARTIFICIAL CHROMOSOME CONTIG MAP OF CHROMOSOME 12

酵母 12 号染色体人工染色体重叠群图谱

• 批准号: 6241459
• 负责人: Raju S. Kucherlapati
• 金额: $ 10万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-08 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6241459
• 关键词: DNA; artificial chromosomes; chromosomes; computer assisted sequence analysis; cytogenetics; fluorescent dye /probe; fungal genetics; genetic library; genetic mapping; genetic markers; genome; human genetic material tag; human tissue; in situ hybridization; molecular cloning; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; pulsed field gel electrophoresis; sequence tagged sites

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.

项目2的目标是开发一个高分辨率的序列标记， 位点（STS）的内容为基础的12号染色体的地图。 我们提出一个方案， 与丹尼尔科恩博士和他的同事们密切合作， 巴黎CEPH的同事们正在开发一种低分辨率的物理 整个人类基因组的图谱。 我们将获得一个12号染色体特异性的 大插入片段酵母人工染色体（YAC）亚文库 最近在巴黎建成的图书馆。 此外，科恩博士将 为我们提供了12号染色体上YAC的信息 已通过指纹分析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活动中心的规模 在这些重叠群中，从每个YAC产生末端特异性序列。 确定为源自12号染色体的所有末端序列 将被排序，至少部分地，以生成STS。 每一个YAC 将筛选重叠群内的每一个的存在或不存在 STSs。 所获得的二进制信息将用于构建STS- 内容地图 我们建议利用同源重组 构建极高分辨率地图的方法，其中 将推导出STS之间的精确顺序和距离。 我们提出了一个 结合各种方法，找出差距并加以填补， 12号染色体的物理图谱。 除了YAC端特定STS之外， 我们将使用所有的遗传标记， 正在由几个小组在构建12号染色体时开发 物理地图 这个过程使我们能够整合基因图谱 在其他地方用我们将要构建的物理地图来构建。 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，以确定区域 高或低重组。 为了填补基因图谱的空白，我们 提出一个定向的努力，以分离微卫星含有序列 从特定的YAC中提取并使用它们来分型CEPH谱系。 我们的物理 测绘工作将补充并建立在CEPH全球工作的基础上 和其他机构，并可以提供一个新的范式， 高分辨率的人类染色体图谱。