Characterization of the human and other model genomes requires the ability to establish the functional significance of DNA sequences being cloned and organized during the intensive genetic and physical mapping studies that are ongoing. One approach is to identify the structural genes contained within this complex collection of information. The development of exon amplification allows for rapid and efficient identification and cloning of coding sequences from complex sources of mammalian genomic DNA. These sequences are isolated by virtue of selection for both functional 5' and 3' splice sites in the genomic sequence being analyzed. Recent tests of modifications to this method indicate exon amplification is now a rapid, highly reliable approach to gene identification and is presently adaptable to large scale analyses. The experimental approaches outlined in this proposal aim to 1) construct and organize a human chromosome 9 specific exon library and 2) develop a high resolution gene map of the 9q34 subregion. The isolation of chromosome 9 specific exon sequences will aid in physical and gene map construction of this chromosome, and in identification of the gene from which the exon originated. The results of these studies will permit development an approach for similar analyses of entire chromosomes. The construction of gene maps of mammalian genomes will provide a new framework for the study of structural, functional, and organizational aspects of chromosomes, and will facilitate more rapid genomic sequencing. The specific availability of a high resolution gene map for 9q34 is expected to accelerate the cloning and characterization of several disease genes that have been mapped to human chromosome 9q34 such as tuberous sclerosis, nail patella syndrome, and a form of torsion dystonia.