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.
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