PHYSICAL MAP OF CHROMOSOME 5

该提案的长期目标是开发一个高 分辨率，人类 5 号染色体的连续物理图。该图将 使用几种不同但互补的方法构建， 包括：自然删除映射，它将细分 染色体分为 30 个独立的区室；辐射混合测绘 将提供整个染色体的有序且连续的图谱 至少 500 Kb 级别的分辨率；原位多色荧光 杂交，当辐射时将提供顺序信息 混合映射不能，最后，建立许多 酵母人工产物形式的克隆 DNA 的重叠片段 染色体 (YAC) 和粘粒重叠群，每个重叠群至少跨越 不同地区的 200 万个碱基对。 提出的一种方法是 实现后一个目标是一种利用辐射混合的新颖方法 从流式排序的 5 号染色体中开发区域特异性子文库 科斯米德图书馆。 物理地图上至少会放置 500 个标记，包括 至少100个高度多态性遗传标记和超过100个已知基因 功能。 每个标记将采用寡核苷酸引物的形式 用于 PCR（序列标记位点，或 STS）。 放置的遗传标记 物理地图将有助于将物理地图与 减数分裂或连锁图。 将两种地图关联起来可以提供 立即提取重要的生物和临床信息的强大方法 来自染色体的信息。 特别是，整合遗传和 物理图谱对于将图谱与疾病表型联系起来至关重要 与至少七种遗传性疾病有关，其基因 已知位于 5 号染色体上。此外，了解 已知功能的基因在物理图上的精确位置可以 对于评估他们作为参与的候选人来说是无价的 遗传性或获得性疾病，包括各种类型的白血病。

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.