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Molecular and cytological characterization of repetitive DNA sequences from the centromeric heterochromatin of Sciara coprophila
Authors:M. Carmen Escribá  Patricia G. Greciano  María Méndez-Lago  Beatriz de Pablos  Vladimir A. Trifonov  Malcolm A. Ferguson-Smith  Clara Goday  Alfredo Villasante
Affiliation:1.Centro de Biología Molecular “Severo Ochoa” (CSIC-UAM),Universidad Autónoma de Madrid,Madrid,Spain;2.Departamento de Proliferación Celular y Desarrollo,Centro de Investigaciones Biológicas,Madrid,Spain;3.Department of Veterinary Medicine,Cambridge Resource Centre for Comparative Genomics,Cambridge,UK;4.Molecular and Cellular Biology Department,Institute of Chemical Biology and Fundamental Medicine,Novosibirsk,Russia;5.Department of Surgery, Committee on Cell Physiology, and Committee on Molecular Pathogenesis and Molecular Medicine,The University of Chicago,Chicago,USA;6.Genome Sciences Center,British Columbia Cancer Research Center,Vancouver,Canada
Abstract:Sciara coprophila (Diptera, Nematocera) constitutes a classic model to analyze unusual chromosome behavior such as the somatic elimination of paternal X chromosomes, the elimination of the whole paternal, plus non-disjunction of the maternal X chromosome at male meiosis. The molecular organization of the heterochromatin in S. coprophila is mostly unknown except for the ribosomal DNA located in the X chromosome pericentromeric heterochromatin. The characterization of the centromeric regions, thus, is an essential and required step for the establishment of S. coprophila as a model system to study fundamental mechanisms of chromosome segregation. To accomplish such a study, heterochromatic sections of the X chromosome centromeric region from salivary glands polytene chromosomes were microdissected and microcloned. Here, we report the identification and characterization of two tandem repeated DNA sequences from the pericentromeric region of the X chromosome, a pericentromeric RTE element and an AT-rich centromeric satellite. These sequences will be important tools for the cloning of S. coprophila centromeric heterochromatin using libraries of large genomic clones.
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