HUMAN DISEASE ISOLATION USING GENETIC LINKAGE AND MOLECULAR CLONING TECHNIQUES

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The long term objective of this research is to develop a general approach for isolating any human gene corresponding to a genetic disease locus. Ultimately these approaches should allow the isolation of large numbers of genes including those for cystic fibrosis, manic depression, Wilson's disease, Polysistic Kidney disease and a host of other similar diseases. Once the genes have been identified the exact genetic and biochemical nature of the disease can be determined. From this understanding a much more directed investigation can be made for early detection and early treatment. The general approach is to use the recently develop restriction fragment length polymorphism (RFLP) map of the human to first locate a disease locus, we then will develop a new yeast artificial chromosome vector for cloning the DNA between two flanking RFLP. The disease locus will then be contained within the cloned DNA. The amount of DNA will approximately 1 to 5 million basepairs (Mb) and will probably contain a number of normal of normal genes with the defective gene. To narrow the search defective gene a high resolution RFLP map will be generated by a new physical mapping procedure an a fine structure recombination and linkage disequilibrium map will be prepared by surveying families transmitting the disease. From this high resolution map the gene should be closely pinpointed. Messenger RNA,s will be surveyed in this area and the exact gene will be determined by direct DNA sequencing.