The Human Genome Initiative has as its first major goal the construction of a complete physical and genetic map of the human genome with the ultimate aim of determining its complete sequence. We propose to establish a Genome Center dedicated to generating a map of human Chromosome 22g. A consortium of 27 experienced investigators at the Children's Hospital of Philadelphia, the University of Pennsylvania, Fox-Chase Cancer Center, DuPont, and Unisys constitute the Center that supports 5 highly integrated projects, 5 technical cores, and 1 administrative core, dedicated to constructing detailed physical and genetic maps of chromosome 22q. The Center has the following specific aims: (1) Isolate a set of >300 anchor markers from chromosome 22q that consist of transcribed sequences, highly polymorphic sequences, and sequences containing rare enzyme cutting sites, thereby insuring with >99% probability that there is at least one anchor marker every 1 mb; (2) Generate sufficient DNA sequence from each anchor marker and design oligonucleotide primers suitable for the polymerase chain reaction in order to establish each marker as a sequence tagged site (STS) ; (3a) Map physically the anchor markers first at low resolution into groups using a panel of somatic cell hybrids and then at higher resolution within each group by using radiation hybrids, in-situ hybridization with fluorescent probes, and pulsed field gel electrophoresis; (3b) Map polymorphic probes genetically on the extended CEPH pedigrees,, (4a) Make large insert genomic libraries of human DNA from chromosome 22 in yeast artificial chromosome and PI vectors, from somatic cell hybrids with only human 22 and/or flow-sorted chromosome 22 material, and use the anchor markers as starting points for constructing overlapping contigs of large genomic clones; (4b) Generate additional STSs within growing contigs at approximately 100 kb intervals; (5) Evaluate and solve the information handling demands of the Human Genome Initiative in general as well as the more immediate needs of the Chromosome 22 Project in particular by combining to best advantage the strengths of sound software engineering practices with the flexibility of an advanced data analysis and management informatics research effort. The goal is to have the entire 22q isolated in overlapping contigs that are anchored by regions containing transcribed and/or polymorphic sequences. A Human Genome Center will provide an integrated, cohesive framework within which a large and dedicated group of investigators can successfully pursue a complete map of a human chromosome.
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