DNA SEQUENCING, TESTING THE OSS APPROACH AND RELATED TECHNOLOGY

基于yac的7号染色体重叠克隆/STS/遗传连锁图谱

A YAC-based overlapping clone/STS/genetic linkage map of chromosome 7 will be constructed. Primer pairs will be developed from the sequence of 200 to 1000 bp clones of chromosome specific DNA, obtained from flow-sorted chromosomes, and will be used in PCR reactions to screen a library of human DNA in YACs for cognate clones. Overlapping YACs will be used to obtain contigs of an average length at least 2 Mb aligned by their probe content. Subsidiary efforts will be made to identify YACs and build contigs from existing probes in regions of current interest on chromosome 7 (e.g., cloned genes). Closure of the map, and its merger with genetic and cytogenetic data, will be based first on the generation of clones from the ends of YAC contigs, and identification of polymorphisms in these regions of DNA. End-clone polymorphisms will be genetically mapped using crossovers in the chromosomes of the offspring from a primary set of reference pedigrees, which should allow rapid alignment of the corresponding contigs and placement of the loci within the genetic linkage map. Closure of the map will be attained by mapping polymorphisms derived from telomeric YACs and cloned DNAs from the pericentromeric region, and by the use of jumping clones, in situ hybridization and pulsed-field gel mapping. The result will be a map in which cytogenetic assignments, linkage information for genetic loci, and the set of overlapping clones that fully describe chromosome 7 are integrated.