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Isolation and regional localisation of DNA sequences from a human chromosome 11-specific cosmid library
Authors:Roy B Wadey  Peter F R Little  Jon Pritchard  John K Cowell
Institution:(1) Imperial Cancer Research Fund Laboratory of Molecular Genetics in the Department of Haematology and Oncology, Institute of Child Health, 30 Guilford Street, WC1N 1EH London, UK;(2) Department of Biochemistry, Imperial College of Science, Technology and Medicine, Exhibition Road, SW7 London, UK
Abstract:Summary A cosmid library has been prepared in the lorist-B vector from a mouse/human somatic cell hybrid containing region 11q23-11pter as the only human component. This chromosome region is stably maintained in the hybrid as a result of translocation onto one copy of mouse chromosome 13. Individual cosmids containing human DNA were isolated by their ability to hybridise with total human DNA, digested with either HindIII or EcoRI, and 33 individual unique sequences were identified. These fragments were then isolated and subcloned into the bluescribe plasmid vector. Regional localisation of these unique sequences was achieved using a panel of somatic cell hybrids containing different overlapping deletions of chromosome 11. The majority of the 33 mapped sequences derived from the long arm of chromosome 11. Two clones were located within the 11p13–p14 region, which is associated with a predisposition to Wilms' tumour. These probes supplement those already mapped to this chromosome and will assist in the generation of a detailed chromosome 11 linkage map.
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