title: Multicolor FISH mapping with Alu-PCR-amplified YAC clone DNA determines the order of markers in the BRCA1 region on chromosome 17q12-q21.
abstract: A gene designated BRCA1, implicated in the susceptibility to early-onset familial breast cancer, has recently been localized to chromosome 17q12-q21. To date, the order of DNA markers mapped within this region has been based on genetic linkage analysis. We report the use of multicolor fluorescence in situ hybridization to establish a physically based map of five polymorphic DNA markers and 10 cloned genes spanning this region. Three cosmid clones and Alu-PCR-generated products derived from 12 yeast artificial chromosome clones representing each of these markers were used in two-color mapping experiments to determine an initial proximity of markers relative to each other on metaphase chromosomes. Interphase mapping was then employed to determine the order and orientation of closely spaced loci by direct visualization of fluorescent signals following hybridization of three probes, each detected in a different color. Statistical analysis of the combined data suggests that the order of markers in the BRCA1 region is cen-THRA1-TOP2-GAS-OF2-17HSD-248yg9-RNU 2-OF3-PPY/p131-EPB3-Mfd188- WNT3-HOX2-GP3A-tel. This map is consistent with that determined by radiation-reduced hybrid mapping and will facilitate positional cloning strategies in efforts to isolate and characterize the BRCA1 gene.
authors.affiliation: University of Michigan, Human Genome Center, Ann Arbor 48109.
authors.forename: W L
authors.initials: WL
authors.lastname: Flejter
authors.forename: C L
authors.initials: CL
authors.lastname: Barcroft
authors.forename: S W
authors.initials: SW
authors.lastname: Guo
authors.forename: E D
authors.initials: ED
authors.lastname: Lynch
authors.forename: M
authors.initials: M
authors.lastname: Boehnke
authors.forename: S
authors.initials: S
authors.lastname: Chandrasekharappa
authors.forename: S
authors.initials: S
authors.lastname: Hayes
authors.forename: F S
authors.initials: FS
authors.lastname: Collins
authors.forename: B L
authors.initials: BL
authors.lastname: Weber
authors.forename: T W
authors.initials: TW
authors.lastname: Glover
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods; D018244:Chromosomes, Artificial, Yeast; D002886:Chromosomes, Human, Pair 17; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005260:Female; D005819:Genetic Markers; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007399:Interphase; D008677:Metaphase; D009857:Oncogenes; D016133:Polymerase Chain Reaction
chemical_list: D005819:Genetic Markers; D004247:DNA