Complete high-resolution genetic (1-2 cM) and physical (1-2 Mb) maps of the human genome are now being constructed. Toward that end, we propose development of a high-resolution physical map for human chromosome 17 by a combination of cytogenetic, somatic cell genetic, and pulsed-field gel mapping strategies. This chromosome comprises 3% of the human genome, representing about 100 Mb. A somatic cell hybrid mapping panel already divides this chromosome into approximately 20 regions, and over 100 cloned genes and polymorphic markers have been mapped on this chromosome. The selectable marker thymidine kinase at 17q22-23 makes somatic cell strategies especially useful for genome analysis. Specific goals of the project include: 1) Expansion of the hybrid mapping panel by: a) isolation of cytogenetically well-defined patient breakpoints in TK- rodent cells to divide the chromosome into -40 regions. This will serve as a reference map for radiation induced breakpoints and pulsed-filed mapping. b) Production of additional random breakpoints (-100) by irradiation of a 17 only hybrid. WE will attempt to rapidly screen and characterize clones by two novel approaches: Alu-PCR to detect presence and amount of human DNA and biotin in situ hybridization of Alu-PCR products or total hybrid DNA as probe against human metaphase chromosomes to localize human segments. 2) Mapping of existing NotI linking clones and construction of additional rare-cutting enzyme linking clones to be used as anchor points of the map. 3) Pulsed-field gel construction of long-range restriction map of the entire chromosome using cloned genes, anonymous RFLP probes, and linking clones which have been previously mapped to defined physical intervals by hybrid analysis. 4) For all cloned genes and conserved sequences (including linking clones), synteny in the mouse on chromosome 11 will be tested using mouse-rat somatic cell hybrids and placed on the mouse genetic map through collaborative arrangements.
