PHYSICAL MAP OF CHROMOSOME 5

这项提议的长期目标是发展一种高 分辨率，连续的人类5号染色体物理图谱。该图谱将 使用几种不同但互补的方法来构建， 包括：自然删除映射，这将细分 染色体分成30个独立的区段；辐射杂交作图 将提供整个染色体的有序和连续的图谱 至少500KB级别的分辨率；多色原位荧光 杂交，它将在辐射时提供顺序信息 混合测绘不能，最后，建立许多 人工酵母形式的克隆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 两百万个碱基对分布在不同的地区。提出了一种方法来 实现后一个目标是一种新的方法，使用辐射混合体来 从流动排序的5号染色体发育区域特异性亚卵泡 COSMID图书馆。 将在实物地图上放置至少500个标记，包括在 至少100个高度多态的遗传标记和100多个已知基因 功能。每个标记都将以寡核苷酸引物的形式出现 用于聚合酶链式反应(序列标签位点，或STSS)。放置在基因上的遗传标记 物理地图将用于将物理地图与 减数分裂或连锁图谱。将这两种地图关联起来提供了一个 立即提取重要生物学和临床重要信息的有效方法 来自染色体的信息。特别是，将遗传和 物理图谱是将图谱与疾病表型联系起来的关键 与至少七种遗传性疾病有关，它们的基因 已知位于5号染色体上。此外，了解到 在物理地图上精确定位已知功能的基因可以 对于评估他们作为候选人参与到 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.