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Nucleic Acids Research, 1988, Vol. 16, No. 19 9185-9198
© 1988

Articles

Construction of a NotI linking library and isolation of new markers close to the Huntington's disease gene

T.M. Pohl, M. Zimmer1,*, M.E. MacDonald2, B. Smith3, M. Bucan1, A. Poustka4, S. Volinia1, S. Searle1, G. Zehetner1, J.J. Wasmuth3, J. Gusella2, H. Lehrach1 and A.-M. Frischauf1

European Molecular Biology Laboratory Postfach 1022.40, D-6900 Heidelberg, FRG 1Imperial Cancer Research Fund, Lincoln's Inn Fields PO Box 123, London, WC2A 3PX, UK 2Neurogenetics Laboratory, Massachusetts General Hospital 32 Fruit Street, Boston, MA 02114 3Department of Biological Chemistry, California College of Medicine, University of California Irvine, CA 92717, USA 4Zentrum fur Molekularbiologie Heidelberg (ZMBH), Im Neuenheimer Feld D-6900 Heidelberg, FRG

Received July 6, 1988. Accepted September 2, 1988.

Linking clones contain sequences flanking recognition sites for enzymes cutting rarely in mammalian DNA. They can be used to obtain and correlate both physical and genetic mapping information over subregions of mammalian chromosomes. We have constructed and used a NotI linking clone library representing unmethylated NotI sites from HHW693 DNA, a hamster hybrid cell line containing 4pl5-4pter and a fragment of 5p as its only human chromosome contribution. Human clones were identified by hybridisation with a cloned human repeat sequence, and localised further to subregions of human chromosome 4pl5-4pter using a panel of additional hybrids. Clones from the region distal to the DNA probes (D4S10, D4S43, D4S95) linked to the Huntington's disease mutation, were further analysed. Four markers close to the HD gene: D4S111, D4S113, D4S114 and clone 417 are described here.

In addition to serving as markers in physical and genetic mapping experiments, these linking clones provide probes next to cleavable NotI sites, and can therefore be used to screen NotI based chromosome jumping libraries. They also provide indications for potential gene sequences, identifiable as evolutionarily conserved sequences.

*Present address: The Salk Institute, PO Box 85800, San Diego, CA 92138-9216, USA


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