Cloning a fragment from the telomere of the long arm of human chromosome 9 in a YAC vector |
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Authors: | Anna Maria Guerrini Fiorentina Ascenzioni Giulio Pisani Giancarlo Rappazzo Giuliano Della Valle Pierluigi Donini |
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Institution: | (1) Dipartimento di Biologia Cellulare e dello Sviluppo, Sezione di Scienze Microbiologiche, Universitá La Sapienza , Via degli Apuli 1, I-00185 Roma, Italy;(2) Medizinisch-Naturwissenschaftliches Forschungszentrum, Universität Tuebingen, D-7400 Tuebingen, Germany;(3) Dipartimento di Biologia Animale, Universitá di Catania, Catania, Italy;(4) Dipartimento di Genetica e Microbiologia A. Buzzati Traverso , Universitá di Pavia, Pavia, Italy |
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Abstract: | The construction of a yeast artificial chromosome containing a human DNA insert is reported. This molecule of about 200 kb behaves as a native yeast chromosome since it has a very high mitotic stability and is present in the yeast transformant clone at a copy number similar to that of the resident chromosomes. Hybridization with the TTAGGG sequence demonstrates that this chromosome contains human telomeric sequences. In situ hybridization of the biotin-labelled artificial chromosome to metaphase human chromosomes shows that the insert occupies a telomeric position on the long arm of chromosome 9. Since the fragment was cloned as an EcoRI insert and not as a telomere, it is situated medially to the telomeric sequences and harbours telomere-associated sequences, that have been shown to contain the TTAGGG sequence. The fragment represents the end of the genetic map of chromosome 9 and thus can be used to characterize the sequence and the structure of the chromosomal region that runs from the end of the chromosome to the first gene. |
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