title: BAC-probes applied for characterization of fragile sites (FS).
abstract: Genomic instability tends to occur at specific genomic regions known as common fragile sites (FS). FS are evolutionarily conserved and generally involve late replicating regions with AT-rich sequences. The possible correlation between some FS and cancer-related breakpoints emphasizes on the importance of understanding the mechanisms of chromosomal instability at these sites. Although about 230 FS have already been mapped cytogenetically, only a few of them have been characterized on a molecular level. In this chapter, we provide a protocol for mapping of common FS using bacterial artificial chromosome (BAC) probes in fluorescence in situ hybridization (FISH) and suggest the usage of lymphocytes from Fanconi anemia patients as a model system. In the latter, rare FS are expressed much more frequently than in, for example, aphidicolin-induced blood lymphocyte preparations. Knowing the exact location of FS enables the molecular comparison of their location and breakpoints that appear during evolution, cancer development and inherited disorders.
authors.affiliation: Jena University Hospital, Institute of Human Genetics, Friedrich Schiller University, Kollegiengasse 10, Jena, D-07743, Germany.
authors.forename: Kristin
authors.initials: K
authors.lastname: Mrasek
authors.forename: Kathleen
authors.initials: K
authors.lastname: Wilhelm
authors.forename: Luciana G
authors.initials: LG
authors.lastname: Quintana
authors.forename: Luise
authors.initials: L
authors.lastname: Theuss
authors.forename: Thomas
authors.initials: T
authors.lastname: Liehr
authors.forename: Andreja
authors.initials: A
authors.lastname: Leskovac
authors.forename: Jelena
authors.initials: J
authors.lastname: Filipovic
authors.forename: Gordana
authors.initials: G
authors.lastname: Joksic
authors.forename: Ivana
authors.initials: I
authors.lastname: Joksic
authors.forename: Anja
authors.initials: A
authors.lastname: Weise
mesh_terms: D016590:Aphidicolin / Q000633:toxicity; D002460:Cell Line; D043283:Chromosome Fragile Sites; D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial / Q000737:chemistry* / Q000235:genetics; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015894:Genome, Human; D042822:Genomic Instability; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008214:Lymphocytes / Q000737:chemistry / Q000187:drug effects / Q000473:pathology; D015335:Molecular Probes / Q000737:chemistry* / Q000235:genetics; D009153:Mutagens / Q000633:toxicity
chemical_list: D015335:Molecular Probes; D009153:Mutagens; D016590:Aphidicolin