HIGH RESOLUTION PHYSICAL MAP OF CHROMOSOME 5

5 号染色体高分辨率物理图谱

• 批准号: 3735834
• 负责人: JOHN J WASMUTH
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3735834
• 关键词: artificial chromosomes; chromosome deletion; chromosomes; genetic library; genetic mapping; genetic markers; human genetic material tag; human tissue; in situ hybridization; linkage mapping; molecular cloning; polymerase chain reaction; pulsed field gel electrophoresis; radiation genetics; restriction fragment length polymorphism; sequence tagged sites

The long term objective of this proposal is to develop a high resolution, contiguous physical map of human chromosome 5. The map will be constructed using several different but complementary approaches, including: natural deletion mapping, which will subdivide the chromosome into 30 separate compartments; radiation hybrid mapping which will provide an ordered and contiguous map of the entire chromosome of at least a 500 Kb level of resolution; multicolor fluorescence in situ hybridization, which will provide order information when radiation hybrid mapping cannot and, finally, the establishment of many overlapping segments of cloned DNA in the form of yeast artificial chromosome (YAC) and cosmid contigs, each of which will span at least two million base pairs in different regions. One method proposed to achieve this latter goal is a novel approach using radiation hybrids to develop region-specific sublibraries from a flow-sorted chromosome 5 cosmid library. AT least 500 markers will be placed on the physical map, including at least 100 highly polymorphic genetic markers and over 100 genes of known function. Each marker will be in the form of oligonucleotide primers for PCR (sequence tagged sites, or STSs). The genetic markers placed on the physical map will serve to integrate the physical map with the meiotic or linkage map. Relating the two kinds of maps provides a powerful way to immediately extract important biological and clinical information from the chromosome. In particular, integrating genetic and physical maps is critical for relating the maps to disease phenotypes associated with at least seven inherited disorders, the genes for which are known to be on chromosome 5. In addition, a knowledge of the precise location of genes of known function on the physical map can be invaluable for assessing them as candidates for being involved for inherited or acquired disorders, including various types of leukemia.

该提议的长期目标是发展高 分辨率，人类染色体5的连续物理图。 可以使用几种不同但互补的方法构建， 包括：自然删除映射，将细分 染色体成30个单独的隔间；辐射混合映射 将提供整个染色体的有序且连续的地图 至少有500 kb的分辨率；多色荧光原位 杂交，该杂交将在辐射时提供订单信息 混合映射不能，最后，建立了许多 酵母人造形式的克隆DNA的重叠段 染色体（YAC）和宇宙重叠群，每个重叠群至少将跨越 不同地区的两百万个碱基对。 提出的一种方法 实现后一个目标是一种使用辐射杂种的新方法 从流量排序的5号染色体开发区域特定的sublibraries 宇宙库。 至少将500个标记放在物理图上，包括 至少100个高度多态遗传标记和已知的100多个基因 功能。 每个标记将以寡核苷酸引物的形式 对于PCR（序列标记的位点或STSS）。 遗传标记 物理地图将用于将物理图与 减数分裂或连锁图。 关联两种地图提供了 立即提取重要生物学和临床的有力方法 来自染色体的信息。 特别是整合遗传和 物理图对于将地图与疾病表型联系起来至关重要 与至少七个遗传疾病相关，这些疾病的基因 已知在5号染色体上。此外， 已知功能基因在物理图上的精确位置可以是 评估他们作为参与的候选人的宝贵 继承或获得的疾病，包括各种类型的白血病。