title: A strategy for the characterization of minute chromosome rearrangements using multiple color fluorescence in situ hybridization with chromosome-specific DNA libraries and YAC clones.
abstract: The identification of marker chromosomes in clinical and tumor cytogenetics by chromosome banding analysis can create problems. In this study, we present a strategy to define minute chromosomal rearrangements by multicolor fluorescence in situ hybridization (FISH) with "whole chromosome painting" probes derived from chromosome-specific DNA libraries and Alu-polymerase chain reaction (PCR) products of various region-specific yeast artificial chromosome (YAC) clones. To demonstrate the usefulness of this strategy for the characterization of chromosome rearrangements unidentifiable by banding techniques, an 8p+ marker chromosome with two extra bands present in the karyotype of a child with multiple anomalies, malformations, and severe mental retardation was investigated. A series of seven-color FISH experiments with sets of fluorochrome-labeled DNA library probes from flow-sorted chromosomes demonstrated that the additional segment on 8p+ was derived from chromosome 6. For a more detailed characterization of the marker chromosome, three-color FISH experiments with library probes specific to chromosomes 6 and 8 were performed in combination with newly established telomeric and subtelomeric YAC clones from 6q25, 6p23, and 8p23. These experiments demonstrated a trisomy 6pter-->6p22 and a monosomy 8pter-->8p23 in the patient. The present limitations for a broad application of this strategy and its possible improvements are discussed.
authors.affiliation: Institut für Humangenetik und Anthropologie der Universität Heidelberg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Popp
authors.forename: A
authors.initials: A
authors.lastname: Jauch
authors.forename: D
authors.initials: D
authors.lastname: Schindler
authors.forename: M R
authors.initials: MR
authors.lastname: Speicher
authors.forename: C
authors.initials: C
authors.lastname: Lengauer
authors.forename: H
authors.initials: H
authors.lastname: Donis-Keller
authors.forename: H C
authors.initials: HC
authors.lastname: Riethman
authors.forename: T
authors.initials: T
authors.lastname: Cremer
mesh_terms: D000015:Abnormalities, Multiple / Q000235:genetics; D002648:Child; D002869:Chromosome Aberrations / Q000235:genetics*; D025063:Chromosome Disorders; D018244:Chromosomes, Artificial, Yeast; D002896:Chromosomes, Human, Pair 6; D002898:Chromosomes, Human, Pair 8; D002999:Clone Cells; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D005260:Female; D015723:Gene Library; D015321:Gene Rearrangement; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D009006:Monosomy; D014314:Trisomy
chemical_list: D015342:DNA Probes; D004247:DNA